CN101517074A - 蛋白酶筛选方法及由此鉴别的蛋白酶 - Google Patents
蛋白酶筛选方法及由此鉴别的蛋白酶 Download PDFInfo
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- CN101517074A CN101517074A CNA2007800328584A CN200780032858A CN101517074A CN 101517074 A CN101517074 A CN 101517074A CN A2007800328584 A CNA2007800328584 A CN A2007800328584A CN 200780032858 A CN200780032858 A CN 200780032858A CN 101517074 A CN101517074 A CN 101517074A
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Abstract
本发明提供了鉴别具有修饰的底物特异性或者其它性质的修饰的蛋白酶的方法。所述方法通过将候选和修饰的蛋白酶与底物接触，当底物裂解时通过形成稳定的复合物而捕获所述蛋白酶，由此对候选和修饰的蛋白酶进行筛选，所述底物例如是丝氨酸蛋白酶抑制蛋白、α巨球蛋白或者p35家族蛋白或者修饰的丝氨酸蛋白酶抑制蛋白和修饰的p35家族成员或者修饰的α巨球蛋白。本发明还提供了修饰的蛋白酶。
Description
相关申请
要求Edwin Madison 2006年7月5日提交的名称为“Protease ScreeningMethods and Proteases Identified Thereby”的美国临时申请系列号No.60/818,804以及Edwin Madison 2006年7月5日提交的名称为“ModifiedUrinary-Plasminogen Activator(u-PA)Proteases”的优先权。当允许时，上述申请的主题以其全文引入本文。
本申请与Edwin Madison 2007年7月5日提交的名称为“ProteaseScreening Methods and Proteases Identified Thereby”的美国申请系列号11/825,627相关，其要求美国临时申请系列号60/818,804和美国临时申请系列号60/818,910的优先权。
本申请还与J Nguyen，C Thanos，S Waugh-Ruggles，和C Craik 2003年10月2日提交的名称为“Methods of Generating and Screening for Proteaseswith Altered Specificity”的美国申请系列号10/677,977并在2004年7月29日公开为美国申请号US-2004-0146938相关以及与2004年4月15日公开的相应公开的国际PCT申请号WO2004/031733相关，其要求2002年10月2日提交的美国临时申请系列号60/415,388的优先权。
本申请还与J Nguyen和S Waugh-Ruggles 2005年4月2日提交的名称为“Cleavage of VEGF and VEGF Receptor by Wild-Type and Mutant MTSP-1”的美国申请系列号11/104,110并在2006年1月5日公开为美国申请号US-2006-0002916相关，以及与2005年11月24日公开的相应公开的国际PCT申请号WO2005/110453相关，其要求2004年4月12日提交的美国临时申请系列号60/561,720的优先权。
本申请还与J Nguyen和S Waugh-Ruggles 2005年4月12日提交的名称为“Cleavage of VEGF and VEGF Receptor by Wild-Type and MutantProteases”的美国申请系列号11/104,111并在2006年2月2日公开为美国申请号US-2006-0024289相关，以及与2005年10月27日公开的相应公开的国际PCT申请号WO2005/100556相关，其要求2004年4月12日提交的美国临时申请系列号60/561,671的优先权。
本申请还与Edwin L.Madison 2005年10月21日提交的名称为“Modified Proteases that Inhibit Complement Activation”的美国临时申请系列号60/729,817相关。本申请还与Edwin L.Madison，Jack Nguyen，SandraWaugh Ruggles和Christopher Thanos 2006年10月20日提交的美国申请系列号11/584,776相关，以及与2007年4月26日公开的相应公开的国际PCT申请号WO2007/047995相关，其各自要求美国临时申请号60/729,817的优先权。
当允许时，上述相关申请的主题以其全文引入本文。
在CD盘上的序列表的引用
CD-R上电子形式的序列表在此提交4个拷贝(标为COPY 1，COPY 2，COPY 3，和CRF)，它们的内容以其全文引入本文。每个上述CD盘的计算机可读文件在2007年7月5日创建，是相同的，大小为1809千比特，名称为4902SEQ.PC1.txt.
发明领域
本发明提供了鉴别具有修饰的底物特异性或者其它性质的修饰的蛋白酶的方法。所述方法通过将候选物和修饰的蛋白酶与底物接触而进行筛选，所述底物在底物裂解时捕获所述候选物和修饰的蛋白酶。
背景
蛋白酶是降解蛋白质的酶。由于蛋白酶可以与靶蛋白特异性相互作用并且使其失活或者活化，因此已经将它们用作治疗剂。天然发生的蛋白酶通常不是最佳的治疗剂，因为其不呈现使其适于作为生物治疗剂的特异性、稳定性和/或催化活性(见例如Fernandez-Gacio et al.(2003)Trends in Biotech.21：408-414)。在治疗性质之中，重要的是无免疫原性或者免疫原性降低、对于靶分子的特异性及有限的副作用。天然发生的蛋白酶通常缺乏这些性质的一种或多种。
已经尝试工程化具有改良性质的蛋白酶。这些方法包括：1)合理设计(rational design)，这需要关于蛋白酶的结构、催化机制和分子建模的信息；2)定向进化，这是包括产生蛋白酶的各种突变体库(repertoire)及选择呈现预期特性的那些突变体的方法(Bertschinger et al.(2005)in Phage displayin Biotech.and Drug Discovery(Sidhu s，ed)，pp.461-491)。就前者而言，缺少关于蛋白酶的结构-功能关系的信息限制了合理设计大多数蛋白酶的突变的能力。采用定向进化方法取得有限的成功。
筛选改良的蛋白酶活性通常导致底物选择性丧失，反之亦然。最佳的治疗性蛋白酶应呈现对于靶底物的高度特异性和高度催化效率。由于可利用的选择具有优化的特异性和优化的活性的蛋白酶的方法的效力有限，因此仍需要开发其它蛋白质选择方法。因此，本发明的目的是提供选择具有希望的底物特异性和活性的蛋白酶或者突变体蛋白酶的方法。
发明概述
本发明提供了选择或鉴别具有希望的或预定的底物特异性和活性的蛋白酶或突变体蛋白酶或其催化活性部分的方法。本发明特别提供了蛋白酶筛选方法，所述方法鉴别对于靶底物具有改变的、改良的或者优化的或者其它改变的底物特异性和/或活性的蛋白酶。所述方法可用于例如筛选对于疾病或病症的病因学中涉及的靶底物具有改变的底物特异性和/或活性的蛋白酶。依靠对于靶底物的改变的、典型地增加的特异性和/或活性，在本发明的方法中鉴别或被选择的蛋白酶是在所述靶底物所参与的疾病或病症的治疗中用作试剂或治疗剂的候选物。在实施本发明的方法中，将蛋白酶或其催化活性部分的集合与蛋白酶捕获多肽接触，导致形成蛋白酶捕获多肽与所述集合中的蛋白酶或其催化活性部分的稳定复合物。在一些实例中，所述蛋白酶捕获多肽被修饰为由对于靶底物(例如参与疾病或病症的靶底物)具有预定底物特异性和/或活性的蛋白酶裂解。所述方法可进一步包括筛选所述复合物对于靶底物的裂解序列的底物特异性。在这种实例中，被鉴别或选择的蛋白酶对于靶底物具有改变的活性和/或特异性。在一个实例中，所述稳定复合物是通过被选择的蛋白酶与蛋白酶捕获多肽的共价连接而形成的。所述被选择的蛋白酶或其催化活性部分是从本发明方法提供的复合物中鉴别或选择的。本发明的方法可进一步包括从集合中未复合的蛋白酶成员中分离复合的蛋白酶的步骤。在一个实例中，对所述蛋白酶捕获多肽进行标记以进行检测和分离，所述分离是通过捕获含有可检测的蛋白酶捕获多肽和所述蛋白酶或其催化活性部分的复合物而实现。捕获可以是在悬浮液、溶液或者固体支持物上实现。在固体支持物的捕获情况中，使所述蛋白酶捕获多肽附着于所述固体支持物，这可以在所述蛋白酶捕获多肽与所述蛋白酶或其催化活性部分集合进行接触之前、期间或随后进行。所述固体支持物可包括例如96孔平板的孔。在一些实例中，用生物素标记所述蛋白酶捕获多肽。在其它实例中，可以用His标记对所述蛋白酶捕获多肽进行标记，所述分离可以通过用金属螯合剂例如但非限于硫酸镍(NiSO4)、氯化钴(CoCl2)、硫酸铜(CuSO4)和氯化锌(ZnCl2)进行捕获而实现。所述金属螯合剂可以与固体支持物缀合，例如与珠如琼脂糖珠或磁珠缀合。
在本发明提供的方法中，所述方法可进一步包括扩增分离的复合物中的蛋白酶或其催化活性部分的步骤。在一些实例中，分离的复合物中的蛋白酶或其催化活性部分在噬菌体上展示，扩增是通过用所述噬菌体感染宿主细胞而实现。所述宿主细胞可包括细菌，例如大肠杆菌(E.coli)。可以筛选从细菌细胞培养基、细菌周质、噬菌体上清或者纯化的蛋白质中扩增的蛋白酶的靶底物特异性和/或活性。典型地，所述靶底物是疾病或病症病因学中涉及的多肽或裂解序列。
本发明还提供了多重方法，其中将蛋白酶集合与多种不同的蛋白酶捕获多肽(包括其修饰的形式)接触，对每个蛋白酶捕获多肽均进行标记，由此其可以被可鉴别地检测。在这种方法中，至少两个蛋白酶捕获多肽被可鉴别地标记，由此可以形成一个以上的稳定复合物及鉴别一种以上的蛋白酶。
在本发明提供的方法中，所述方法还包括多轮筛选以优化蛋白酶选择，其中在本发明筛选方法的第一轮中鉴别或选择蛋白酶之后对其进行扩增，从而产生蛋白酶或其催化活性部分的第二个集合。将所述第二个蛋白酶集合与蛋白质捕获多肽接触产生第二组稳定复合物，所述蛋白质捕获多肽与第一种蛋白酶捕获多肽相同或不同。鉴别或选择第二组稳定复合物中的蛋白酶。
在本发明提供的方法中，所述蛋白酶捕获多肽是丝氨酸蛋白酶抑制蛋白，α巨球蛋白家族的成员，或者p35家族的成员。本发明提供的方法中使用的这种多肽分子通过蛋白酶或其催化活性部分与蛋白酶捕获多肽的共价连接而形成复合物。
在本发明提供的方法的一方面，鉴别了具有希望的底物特异性的蛋白酶，通过将蛋白酶和/或蛋白酶的蛋白酶解活性部分的集合与蛋白酶捕获多肽接触，在所述蛋白酶捕获多肽裂解时形成蛋白酶捕获多肽与蛋白酶的稳定复合物。选择蛋白酶捕获多肽或其修饰形式用于所述方法中，以由具有希望的底物特异性的蛋白酶裂解。在所述方法中，鉴别了所述蛋白酶或其蛋白酶解活性部分，以选择具有希望的底物特异性的蛋白酶。
本发明提供的方法中使用的蛋白酶集合是蛋白酶或其催化活性部分的任何集合，包括至少、大约或者等于5、10、50、100、103、104、105、106或更多个成员。在一些情况中，所述蛋白酶是丝氨酸和/或半胱氨酸蛋白酶。在本发明提供的方法中，蛋白酶或其催化活性部分的集合被展示以与蛋白酶捕获多肽接触。在一个实例中，所述蛋白酶或其蛋白酶解活性部分被展示在固体支持物、细胞表面或者微生物表面上。所述蛋白酶可以被展示在酵母、细菌、病毒、噬菌体、核酸、mRNA分子或者核糖体上。在蛋白酶或其蛋白酶解活性部分被展示在微生物上的情况中，所述微生物包括但非限于大肠杆菌、酿酒酵母(S.cerevisiae)，或者病毒如M13、fd或T7噬菌体或者杆状病毒。在本发明提供的方法中，蛋白酶或其蛋白酶解活性部分被展示在噬菌体展示文库上，所述蛋白酶集合是蛋白酶噬菌体展示文库。在一些实施方案中，蛋白酶例如通过被展示为全长蛋白酶的蛋白酶解活性部分而提供在所述集合中。在一些实例中，蛋白酶集合与蛋白酶捕获多肽的接触是在同质混合物中进行。
本发明提供了蛋白酶选择方法，其中将至少两种不同的蛋白酶捕获多肽与所述集合接触，但是只有一种蛋白酶捕获多肽是可检测地标记的。所述可检测地标记的蛋白酶捕获多肽可以捕获含有所述可检测的蛋白酶捕获多肽和蛋白酶或其催化活性部分的稳定复合物。在这种方法的一些实例中，在反应中存在与可检测地标记的蛋白酶捕获多肽相比过量存在的一或多种未被可检测地标记的其它蛋白酶捕获多肽。在所述方法中，所述标记是可以检测的任何标记，例如荧光标记或者表位标记，如His标记。在其它实例中，所述可检测的标记是生物素。
本发明方法中进行选择的蛋白酶的集合包括蛋白酶的任何集合。在一些实例中，所述蛋白酶是丝氨酸或半胱氨酸蛋白酶。所述蛋白酶集合包括是丝氨酸蛋白酶的胰凝乳蛋白酶及枯草杆菌蛋白酶家族成员或者来自半胱氨酸蛋白酶木瓜蛋白酶家族的胱天蛋白酶。所述蛋白酶包括任何蛋白酶或其催化活性部分，如表7示出。在一些实例中，所述蛋白酶或其催化活性部分是尿激酶纤溶酶原激活物(u-PA)蛋白酶、组织纤溶酶原激活物(t-PA)蛋白酶或者MT-SP1蛋白酶。
一方面，本发明方法中使用的蛋白酶捕获多肽是丝氨酸蛋白酶抑制蛋白、p35家族成员、α-巨球蛋白家族成员，或者其任何修饰形式。本发明方法中使用的蛋白酶捕获多肽包括但非限于纤溶酶原抑制剂-1(PAI-1)、抗凝血酶(AT3)或者α2-巨球蛋白，或者其修饰形式。本发明方法中使用的蛋白酶捕获多肽的修饰形式包括在所述蛋白酶捕获多肽的反应位点含有氨基酸置换、缺失或者取代的那些修饰形式。在一些实例中，所述修饰是相应于靶底物的裂解序列的任一或多个氨基酸置换。所述靶底物可以是疾病或病症病因学中涉及的任何蛋白质。靶底物的例子包括但非限于VEGFR、t-PA裂解序列，或者补体蛋白。例如，靶底物包括但非限于VEGFR2或者补体蛋白C2。靶底物的裂解序列包括但非限于SEQ ID NO：389、479和498所示的任何序列。在一些情况中，所述蛋白酶捕获多肽是丝氨酸蛋白酶抑制蛋白，且所述一或多个氨基酸置换是位于丝氨酸蛋白酶抑制蛋白多肽的反应位点环中。所述反应位点环(reactive site loop，RSL)中的一或多个置换包括在P4-P2’任一或多个位置中的那些置换。本发明的方法中使用的这种丝氨酸蛋白酶抑制蛋白的例子是SEQ ID NO：497、499、610和611所示的任何序列。在另一实例中，所述蛋白酶捕获多肽是α2巨球蛋白且所述一或多个氨基酸置换位于所述多肽的诱饵区。在本发明方法中针对修饰的蛋白酶捕获多肽被鉴别或选择的蛋白酶可以根据与非靶底物相比对于靶底物的改变的底物特异性进行筛选或选择。在这些实例中，所述非靶底物包括相应的模板蛋白酶的底物。典型地，被鉴别或选择的蛋白酶的底物特异性增加1.5倍、2倍、5倍、10倍、50倍、100倍、200倍、300倍、400倍、500倍或更多倍。
本发明提供的方法包括鉴别和/或选择具有希望的底物特异性的蛋白酶的方法被重复或者进行多次的重复方法。在这种方法中，在所述方法的第一次重复或者第一轮中可以鉴别多种不同的蛋白酶。在其它实例中，在第一次重复后基于在第一轮重复中选择的被鉴别的蛋白酶产生和制备多种蛋白酶。此外，在一些实例中，将在第一轮或重复和/或在连续轮中鉴别的被选择的蛋白酶的氨基酸序列进行比较，以鉴别热点。热点是在多轮中被认为是修饰的基因座的那些位置，如与野生型或模板蛋白酶相比在所述方法使用的修饰的蛋白酶集合中至少2、3、4、5或更多种被鉴别的蛋白酶中出现的那些位置。
本发明方法中还提供了在鉴别蛋白酶之后制备第二个蛋白酶集合的步骤，其中鉴别的蛋白酶用作模板以在蛋白酶序列或其催化活性部分中产生进一步的突变，由此所述第二个集合的成员(基于)含有具有鉴别的蛋白酶的突变及其它突变的多肽；然后将所述第二个集合与用于分离第一种蛋白酶的蛋白酶捕获剂(protease trap)相同或不同的蛋白酶捕获多肽接触，其中所述蛋白酶捕获剂经修饰以被具有希望的底物特异性的蛋白酶裂解；从复合物集合中鉴别蛋白酶或其蛋白酶解活性部分，由此鉴别的蛋白酶与第一种鉴别的蛋白酶相比对于希望的底物具有较高的活性或特异性。所述第二个集合与鉴别的模板蛋白酶或其蛋白酶解活性部分的氨基酸序列相比可以含有随机或聚焦突变(focused mutation)。聚焦突变包括例如热点位置，如在丝氨酸蛋白酶如u-PA中基于胰凝乳蛋白酶编号的第30、73、89和155位。
在实施本发明的方法中，形成稳定复合物的反应可以通过控制一或多个参数而调节。这种参数是改变反应速率或程度或者反应效率的任何参数，例如但非限于反应时间、温度、pH、离子强度、文库浓度和蛋白酶捕获多肽浓度。
反应可以在存在反应竞争剂的条件下在蛋白酶捕获多肽与蛋白酶或其蛋白酶解活性部分之间进行，从而增强鉴别的蛋白酶或其蛋白酶解活性部分的选择性。所述竞争剂包括例如血清或血浆。如人血清或人血浆，细胞或组织提取物，生物学流体如尿液或血液，纯化或部分纯化的野生型形式(或者其它修饰形式)蛋白酶捕获剂。这种竞争剂的例子是纯化或部分纯化的蛋白酶捕获多肽的野生型形式或者蛋白酶捕获多肽的一或多种特异性变体。
本发明提供了进化或者选择或者鉴别对于至少两种裂解序列具有特异性/选择性和/或活性的蛋白酶或其蛋白酶解活性部分的重复方法。所述方法包括如下步骤：a)将蛋白酶和/或其蛋白酶解活性部分的集合与第一种蛋白酶捕获多肽接触，在所述蛋白酶捕获多肽被所述蛋白酶或其蛋白酶解活性部分裂解时形成含有蛋白酶捕获多肽与所述集合中的蛋白酶或其催化活性部分的稳定复合物，其中所述接触是在存在竞争剂的条件下进行；b)鉴别或者选择与第一种蛋白酶捕获多肽形成复合物的蛋白酶或其蛋白酶解活性部分；c)将与第一种蛋白酶捕获多肽形成复合物的蛋白酶或其蛋白酶解活性部分与第二种蛋白酶捕获多肽在存在竞争剂的条件下接触；及d)鉴别或者选择与第一种蛋白酶捕获多肽形成复合物的蛋白酶或其蛋白酶解活性成分。所述两个裂解序列可以在一个靶底物中或者可以在两个不同的靶底物中。所述鉴别的、选择的或者进化的蛋白酶或其蛋白酶解活性部分对于一或两个不同靶底物中的至少两个不同裂解序列具有底物特异性和/或裂解活性。所述第一种和第二种蛋白酶捕获多肽可以相同或不同。典型地，本发明方法中使用的第一种和第二种蛋白酶捕获多肽是不同的，且均被修饰为由对于不同靶底物具有预定底物特异性的蛋白酶裂解。所述方法可进一步包括重复至少一次步骤a)和b)或者a)-d)，直至对于至少两个识别序列具有希望的或者预定的底物特异性和裂解活性的蛋白酶被分离。底物特异性和裂解活性与模板蛋白酶相比典型地增加。
所述方法中使用的竞争剂包括所述蛋白酶捕获多肽可以与其相互作用的任何物质，其典型地具有比靶蛋白酶低的稳定性。竞争剂包括但非限于血清、血浆、人血清或人血浆、细胞或组织提取物、生物学流体如尿液或血液、纯化或部分纯化的蛋白酶捕获剂的野生型形式及蛋白酶捕获多肽的一或多种特异变体。
本发明还提供了蛋白酶选择方法，包括如下步骤：a)将蛋白酶或其蛋白酶解活性部分的集合与第一种蛋白酶捕获多肽接触，在所述蛋白酶捕获多肽裂解时，所述蛋白酶捕获多肽与所述集合中的任何蛋白酶或其催化活性部分形成共价复合物；b)从未复合的蛋白酶捕获多肽中分离复合的蛋白酶；c)分离或者选择或者鉴别复合的蛋白酶；d)基于被选择的蛋白酶产生蛋白酶或其蛋白酶解活性成分的第二个集合；及e)重复步骤a)-c)，将蛋白酶或其蛋白酶解活性部分的第二个集合与第二种蛋白酶捕获多肽(与第一种蛋白酶捕获多肽不同)接触形成复合物；分离所述复合物；以及分离、选择或者鉴别复合的蛋白酶。所述第一种和第二种蛋白酶捕获多肽可以被修饰为含有两个不同的靶底物识别序列，从而被鉴别的或者选择的蛋白酶对于至少两个识别序列具有特异性和高度裂解活性。这些方法可以重复多次。在这些方法中，可以将所述蛋白酶或其蛋白酶解活性部分的集合与所述第一种和/或第二种蛋白酶捕获多肽在存在竞争剂(见上述)的条件下接触。
在本发明提供的任何蛋白酶选择方法中，所述集合可含有修饰的蛋白酶。所述蛋白酶中的修饰可以是随机或定向修饰或者在多肽的靶区域中的修饰。
本发明还提供了含有蛋白酶和/或其蛋白酶解活性部分及至少一种蛋白酶捕获多肽的组合。所述成分可以是单独提供或者是以混合物提供。所述蛋白酶捕获多肽包括丝氨酸蛋白酶抑制蛋白、p35家族成员、α-巨球蛋白家族成员，其修饰的形式及混合物。
蛋白酶和/或其催化活性部分的集合可以是以溶液或者悬浮液或者固相形式提供，或者另外展示在例如固体支持物(基质材料)上或者于展示文库中，例如但非限于噬菌体展示文库，其中文库成员展示至少蛋白酶的蛋白酶解活性部分。
本发明提供了含有所述组合的试剂盒。典型地，所述试剂盒含有包装的成分，及任选其它试剂和进行所述方法的说明书。
本发明还提供了通过修饰选自第30、73、89和155位(基于胰凝乳蛋白酶编号)的一或多个残基而修饰丝氨酸蛋白酶如u-PA的底物特异性的方法。
本发明还提供了通过本发明的方法鉴别的修饰的蛋白酶。所述修饰的蛋白酶由于鉴别的修饰而呈现改变的底物特异性和/或活性。使用所述选择方法鉴别的任何修饰可以在野生型蛋白酶、其任何等位基因或物种变体或者在所述蛋白酶的任何其它变体中产生。另外，本发明还提供了含有与野生型蛋白酶或其等位基因或物种变体具有80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更高的序列相同性的修饰的蛋白酶，只要本文所述方法中鉴别的修饰存在即可。
这种蛋白酶是修饰的蛋白酶，包括胰凝乳蛋白酶家族中修饰的丝氨酸蛋白酶，如在选自第30、73、89和155位(基于胰凝乳蛋白酶编号)的热点位置中含有一或多个突变的尿纤溶酶原激活物(u-PA)多肽或者其催化活性片段，其中底物特异性被改变。
本发明还提供了在本发明的方法中鉴别的修饰的尿纤溶酶原激活物(u-PA)蛋白酶，其呈现对于疾病或病症的病因学中涉及的靶底物的增加的特异性和/或活性。这种靶底物包括但非限于VEGFR或者组织纤溶酶原激活物(t-PA)底物。因此，本发明还提供了裂解t-PA底物的修饰的丝氨酸蛋白酶或者其催化活性部分。特别地，这种蛋白酶是修饰的尿纤溶酶原激活物(u-PA)多肽，其中所述u-PA多肽或其催化活性部分在选自第21、24、30、39、61(A)、80、82、84、89、92、156、158、159和187位(基于胰凝乳蛋白酶编号)的位置含有一或多个修饰。本发明还提供了这种修饰的u-PA多肽或其催化活性部分，其含有选自F21V、I24L、F30V、F30L、T39A、Y61(A)H、E80G、E82E、E84K、I89V、K92E、K156T、T158A、V159A和K187E的一或多个突变。本发明还提供了这些修饰的u-PA多肽，其中u-PA多肽或其催化活性部分含有两或多个突变，所述突变选自F30V/Y61(a)H、F30V/K82E、F30V/K156T、F30V/K82E/V159A、F30V/K82E/T39A/V159A、F30V/K82E/T158A/V159A、F30V/Y61(a)H/K92E、F30V/K82E/V159A/E80G/I89V/K187E及F30V/K82E/V159A/E80G/E84K/I89V\K187E。
本发明还提供了裂解VEGFR、特别是裂解VEGFR-2的修饰的丝氨酸蛋白酶或其催化活性部分。特别地，本发明提供了修饰的尿纤溶酶原激活物(u-PA)多肽及其催化活性部分，其中所述u-PA多肽或其催化活性部分在选自第38、72、73、75、132、133、137、138、155、160和217位的位置(基于胰凝乳蛋白酶编号)含有一或多个修饰，从而改变对于VEGFR-2或其序列的底物特异性。本发明还提供了这样的多肽，其含有选自V38D、R72G、L73A、L73P、S75P、F132L、G133D、E137G、I138T、L155P、L155V、L155M、V160A和R217C的一或多个突变。这些多肽可进一步包括在第30和/或89位(基于胰凝乳蛋白酶编号)的修饰，如选自F30I、F30T、F30L、F30V、F30G、F30M和I89V的修饰。本发明还提供了含有一或多个突变及在一些情况中含有两或多个突变的这些u-PA多肽或其催化活性部分，所述突变选自L73A/I89V、L73P、R217C、L155P、S75P/I89V/I138T、E137G、R72G/L155P、G133D、V160A、V38D、F132L/V160A、L73A/I89V/F30T、L73A/I89V/F30L、L73A/I89V/F30V、L73A/I89V/F30G、L73A/I89V/L155V、L73A/I89V/F30M、L73A/I89V/L155M、L73A/I89V/F30L/L155M和L73A/I89V/F30G/L155M。
本发明还提供了修饰的u-PA多肽，其中所述u-PA多肽或其催化活性部分含有选自F30I、F30T、F30G和F30M的一或多个突变。
本发明还提供了通过本发明方法鉴别的修饰的MT-SP1多肽。这种修饰的多肽包括具有选自如下的一或多个氨基酸修饰的SEQ ID NO：253所示MT-SP1多肽：D23E、I41F、I41T、L52M、Y60(g)s、T56K、H71R、F93L、N95K、F97Y、F97L、T98P、F99L、A126T、V129D、P131S、I136T、I136V、H143R、T144I、I154V、N164D、T166A、L171F、P173S、F184(a)L、Q192H、S201I、Q209L、Q221(a)L、R230W、F234L和V244G(基于胰凝乳蛋白酶编号)。在一些实例中，所述修饰是在具有SEQ ID NO：505所示氨基酸序列的MT-SP1的催化活性部分中。在其它实例中，所述修饰是在MT-SP1多肽中进一步包含相应于SEQ ID NO：253所示MT-SP1多肽中C122S修饰的修饰(基于胰凝乳蛋白酶编号)，例如SEQ ID NO：507或517所示MT-SP1。本发明特别提供的修饰是选自I41F、F97Y、L171F和V244G的任何修饰。本发明提供的修饰的MT-SP1多肽可进一步包括相应于SEQ ID NO：253所示MT-SP1多肽中Q175R或D217V的一或多个修饰。这种修饰包括选自如下的任何修饰：I136T/N164D/T166A/F184(A)L/D217V、I41F、I41F/A126T/V244G、D23E/I41F/T98P/T144I、I41F/L171F/V244G、H143R/Q175R、I41F/L171F、R230W、I41F/I154V/V244G、I141F/L52M/V129D/Q221(A)L、F99L、F97Y/I136V/Q192H/S201I、H71R/P131S/D217V、D217V、T65K/F93L/F97Y/D217V、I41T/P173S/Q209L、F97L/F234L、Q175R、N95K和Y60(G)S。任何上述修饰的MT-SP1多肽呈现增加对于C2补体蛋白的特异性或者对于C2补体蛋白的活性或者这二者的修饰。
本发明还提供了药物组合物，其含有修饰的蛋白酶，包括修饰的丝氨酸蛋白酶如修饰的u-PA多肽或者修饰的MT-SP1多肽。所述药物组合物含有药物可接受的赋形剂，可以根据任何合适的给药途径而加以配制，所述给药途径包括但非限于系统、口服、经鼻、肺部、局部(local)及表面(topical)给药途径。本发明还提供了试剂盒，所述试剂盒含有任何所述药物组合物、给予所述组合物的装置及任选包含给药说明书。
本发明提供了编码修饰的蛋白酶、包括u-PA蛋白酶和MT-SP1蛋白酶及其催化活性部分的核酸分子。本发明还提供了含有所述核酸分子的载体及含有所述核酸分子或载体的细胞。
本发明提供了治疗患有疾病或病症的对象的方法，所述疾病或病症例如但非限于是动脉血栓形成、静脉血栓形成和血栓栓塞、缺血性脑中风、后天凝血障碍、弥散性血管内凝血、细菌感染和牙周炎及神经系统疾病，其通过给予t-PA治疗，通过给予含有修饰的u-PA蛋白酶或其蛋白酶解部分或者编码核酸分子或者细胞的药物组合物而进行。所述方法通过给予对象核酸分子、细胞或者药物组合物而实现。
本发明还提供了治疗患有由VEGFR、特别是VEGFR-2介导的疾病或病症的对象的方法。这种疾病和病症包括但非限于癌症、血管生成相关疾病、眼病如黄斑变性、炎症疾病和糖尿病，特别是糖尿病并发症如糖尿病视网膜病变。所述方法通过给予含有或者编码修饰的u-PA蛋白酶的核酸分子或者细胞或者组合物实现，所述修饰的蛋白酶与未修饰的形式相比呈现对于VEGFR-2的底物特异性，特别是增加的底物特异性。所述方法任选包括给予治疗所述疾病或病症的另一种药物，例如在所述疾病是癌症时给予抗肿瘤药物。
本发明还提供了治疗患有由补体蛋白、特别是C2介导的疾病或病症的对象的方法。这种疾病或病症包括但非限于败血症、类风湿性关节炎(RA)、膜增生性肾小球肾炎(MPGN)、多发性硬化(MS)、重症肌无力(MG)、哮喘、炎症性肠病、免疫复合物(IC)介导的急性炎症性组织损伤、阿尔茨海默病(AD)、缺血-再灌注损伤，及Guillan-Barre综合征。在一些实例中，所述缺血-再灌注损伤是由于例如但非限于心肌梗塞(MI)、中风、血管成形术、冠状动脉旁路移植术、心肺转流术(CPB)和血液透析等事件或治疗而导致的。所述方法是给予含有或编码修饰的MT-SP1蛋白酶的核酸分子或者细胞或者组合物实现的，所述修饰的蛋白酶与未修饰的形式相比呈现对于C2的底物特异性，特别是增加的底物特异性。在其它实例中，所述疾病或病症由对对象进行的治疗导致。例如，所述治疗可以导致补体介导的缺血-再灌注损伤。这种治疗包括但非限于血管成形术或冠状动脉旁路移植术。在这些实例中，在对对象进行治疗之前给予修饰的MT-SP1蛋白酶。所述修饰的MT-SP1多肽可以通过在体外、离体或者在体内与体液或者组织样品接触而给予。
本发明还提供了本发明提供的方法中使用的修饰的丝氨酸蛋白酶抑制蛋白多肽。这种修饰的丝氨酸蛋白酶抑制蛋白多肽在其反应位点环中相应于第P4-P2’位的位置用靶底物的裂解序列修饰。典型地，所述靶底物参与疾病或病症的病因学。这种靶底物包括但非限于VEGFR、补体蛋白或者t-PA底物。例如，靶底物包括VEGFR2和补体蛋白C2。本发明提供的修饰的丝氨酸蛋白酶抑制蛋白的例子是修饰的纤溶酶原激活物抑制剂-1(PAI-1)和抗凝血酶-3(AT3)。修饰的丝氨酸蛋白酶抑制蛋白例如SEQ ID NO：497、499、610和611所示。
附图简述
图1描述了丝氨酸蛋白酶抑制蛋白对蛋白酶的抑制机制及稳定的抑制复合物的产生。在接触之后，丝氨酸蛋白酶抑制蛋白(I)和蛋白酶(E)最初形成非共价的Michaelis样复合物(EI)。随后是P1-P1’易断裂键的裂解及蛋白酶的催化性丝氨酸对丝氨酸蛋白酶抑制蛋白的反应位点环(RSL)羰基的亲核攻击以及共价的酰基-酶中间体(EI#)的形成。动态捕获的共价抑制产物(EI+)是RSL插入、蛋白酶移位、蛋白酶活性位点变形及整体蛋白酶结构变形的结果。次要的非抑制途径释放正常的裂解产物丝氨酸蛋白酶抑制蛋白(I＊)和反应性蛋白酶(E)。
详细描述
概要
A.定义
B.筛选蛋白酶的方法
C.蛋白酶捕获多肽
1.丝氨酸蛋白酶抑制蛋白：结构、功能和表达
2.蛋白酶催化、丝氨酸蛋白酶抑制蛋白的抑制机制及酰基酶中间体的形成
a.丝氨酸蛋白酶抑制蛋白的例子
i.PAI-1
ii.抗凝血酶(AT3)
3.其它蛋白酶捕获多肽
a.p35
b.α巨球蛋白(aM)
4.蛋白酶捕获多肽竞争剂
5.蛋白酶捕获多肽变体
D.蛋白酶
1.候选蛋白酶
a.蛋白酶的类别
i.丝氨酸蛋白酶
(a)尿激酶型纤溶酶原激活物(u-PA)
(b)组织纤溶酶原激活物(t-PA)
(c)MT-SP1
ii.半胱氨酸蛋白酶
E.进行筛选的修饰的蛋白酶和集合
1.变体蛋白酶的产生
a.随机诱变
b.聚焦诱变(Focused Mutagenesis)
2.变体蛋白酶的嵌合形式
3.组合文库及其它文库
a.噬菌体展示文库
b.细胞表面展示文库
c.其它展示文库
F.接触、分离和鉴别选择的蛋白酶的方法
1.重复筛选
2.选择的蛋白酶的例子
G.评定蛋白酶活性和特异性的方法
H.产生编码蛋白酶捕获多肽(即丝氨酸蛋白酶抑制蛋白)或其变体或者蛋白酶/修饰的蛋白酶的核酸的方法
1.载体和细胞
2.表达
a.原核细胞
b.酵母细胞
c.昆虫细胞
d.哺乳动物细胞
e.植物
3.纯化技术
4.融合蛋白
5.核苷酸序列
I.选择的蛋白酶多肽的制备、配制和给予
1.组合物与输送
2.选择的蛋白酶的体内表达与基因治疗
a.核酸的输送
i.载体-附加型载体与整合载体
ii.人工染色体及其它非病毒载体输送方法
iii.含有核酸的细胞的脂质体及其它胶囊形式和给予
b.体外及离体输送
c.全身、局部和表面输送
3.组合治疗
4.产品和试剂盒的物品
J.使用选择的蛋白酶多肽的治疗方法实例
1.裂解tPA靶位的选择的uPA多肽的治疗方法实例
a.血栓形成性疾病和病症
i.动脉血栓形成
ii静脉血栓形成和血栓栓塞
(a)缺血性脑中风
iii.后天凝血障碍
(a)弥散性血管内凝血(DIC)
(b)细菌感染和牙周炎
b.其它tPA靶位相关病症
c.诊断方法
2.裂解VEGF或VEGFR靶位的选择的蛋白酶多肽的治疗方法实例
a.血管生成、癌症及其它细胞周期依赖性疾病或病症
b.使用裂解VEGF或VEGFR的选择的蛋白酶的组合治疗
3.裂解补体蛋白靶位的选择的MT-SP1多肽的治疗方法实例
a.免疫介导的炎症性疾病
b.神经退行性疾病
c.心血管疾病
K.实施例
A.定义
除非特别指出，本文所用所有技术和科学术语均具有本发明所属领域技术人员熟知的相同含义。除非特别指出，本文中所列举的所有专利、专利申请、公布的申请和出版物、Genbank序列、数据库、网站及其它公布的材料均以其全部内容并入作参考。当本文的术语有多种定义时，本章中的定义起决定作用。当参考URL或其它这种标识符或地址时，应理解这种标识符可以改变并且互联网上的特定信息可以来来去去，但是通过检索互联网可以发现等价信息。其中的引用表明这种信息的可利用性和公共传播。
如本文所用，“蛋白酶捕获剂”或者“蛋白酶捕获多肽”是指由蛋白酶裂解的底物，且在裂解时与所述蛋白酶形成稳定复合物，从而随着所述蛋白酶经历实际的过渡状态形成酶复合物而捕获所述蛋白酶，从而抑制蛋白酶的活性并捕获它。因此，蛋白酶捕获剂是蛋白酶的抑制剂。蛋白酶捕获剂是多肽或分子，其包括由蛋白酶裂解的氨基酸残基，由此当裂解时形成稳定复合物。所述复合物足够稳定，可以允许从未反应的捕获剂或者从与所述蛋白酶较不稳定地相互作用的捕获剂中分离所述复合物。蛋白酶捕获剂包括被所述蛋白酶裂解的合成的、修饰的或者天然发生的任何分子，当裂解时与所述蛋白酶形成复合物，使得可以从未反应的捕获剂中分离出反应的蛋白酶或复合物。这种蛋白酶捕获剂的例子是丝氨酸蛋白酶抑制蛋白，修饰的丝氨酸蛋白酶抑制蛋白，呈现与丝氨酸蛋白酶抑制蛋白相似机制的分子，例如p35，及由蛋白酶裂解并且捕获蛋白酶形成稳定复合物的任何其它分子，例如α2巨球蛋白。蛋白酶捕获剂还包括合成的多肽，其由蛋白酶(或其蛋白酶解活性部分)裂解，当裂解时与所述蛋白酶或其蛋白酶解活性部分形成稳定复合物。
如本文所用，丝氨酸蛋白酶抑制蛋白是指抑制蛋白酶的一组结构相关的蛋白质，当其反应位点由蛋白酶裂解时导致稳定的酰基-酶中间体形成及稳定的共价复合物中的蛋白酶的捕获。丝氨酸蛋白酶抑制蛋白包括等位基因和物种变体及其它变体，只要所述丝氨酸蛋白酶抑制蛋白分子通过形成稳定的共价复合物而抑制蛋白酶。丝氨酸蛋白酶抑制蛋白还包括全长丝氨酸蛋白酶抑制蛋白多肽的截短或连续的氨基酸片段，其最低限度包括至少足够的反应位点环(RSL)部分，以促进蛋白酶抑制及与所述蛋白酶的稳定的共价复合物的形成。丝氨酸蛋白酶抑制蛋白的实例在表2中列出和/或具有SEQ ID NO：1-38任一所示的氨基酸序列、其等位基因变体或者截短部分。
如本文所用，丝氨酸蛋白酶抑制蛋白的“突变体”或者“变体”是指含有氨基酸修饰的丝氨酸蛋白酶抑制蛋白，特别是在丝氨酸蛋白酶抑制蛋白的反应位点环中含有修饰。所述修饰可以是在相应于Pn-P15-P14-P13....P4-P3-P2-P1-P1’-P2’-P3’...Pn’的位置一或多个氨基酸的置换、缺失或者取代。典型地，所述丝氨酸蛋白酶抑制蛋白与野生型丝氨酸蛋白酶抑制蛋白相比在反应位点环中的1、2、3、4、5、6、7、8、9、10或更多个氨基酸位置上含有氨基酸置换。最通常地，所述置换是在相应于P4-P2’位置的一或多个氨基酸上。例如，SEQ ID NO：11所示的PAI-1丝氨酸蛋白酶抑制蛋白，相应于SEQ ID NO：11第366-370氨基酸位置的P4-P1’位置(VSARM)可以被修饰。实施例1描述了VSARM氨基酸残基被修饰为RRVRM或PFGRS。
如本文所用，“易断裂的键”是指被蛋白酶裂解的多肽中的键，表示在底物的裂解序列中P1-P1’位置之间形成的键。
如本文所用，反应位点是指由蛋白酶裂解的那部分靶底物序列。典型地，反应位点包括P1-P1’易断裂键序列。
如本文所用，反应位点环(RSL，也称作反应中心环RCL)是指丝氨酸蛋白酶抑制蛋白分子中的氨基酸序列(典型为17-22个连续氨基酸)，其作为蛋白酶识别位点且通常含有唯一的或者主要的蛋白酶特异性决定簇。RSL序列的裂解及其构象变化是稳定的共价复合物中丝氨酸蛋白酶抑制蛋白分子对蛋白酶的捕获的结果。对于本发明之目的，丝氨酸蛋白酶抑制蛋白的RSL环中的任一或多个氨基酸均可以被修饰为相应于希望的靶蛋白的裂解序列。这种修饰的丝氨酸蛋白酶抑制蛋白或其含有变体RSL序列的部分可以用于选择具有改变的底物特异性的蛋白酶。
如本文所用，分配(partitioning)是指稳定的丝氨酸蛋白酶抑制蛋白-蛋白酶复合物与裂解的丝氨酸蛋白酶抑制蛋白之间丝氨酸蛋白酶抑制蛋白分配的过程。丝氨酸蛋白酶抑制蛋白中的分配的原因与抑制途径的性质有关，其得自裂解的反应位点环经过丝氨酸蛋白酶抑制蛋白表面的大量移位。如果蛋白酶在采取抑制的位置之前有时间解离(即酶-丝氨酸蛋白酶抑制蛋白复合物去酰基)，则发生分配。因此，丝氨酸蛋白酶抑制蛋白-蛋白酶相互作用的结果依赖于抑制(k4)和底物(k5)途径之间的分配比率(如图1示出)，其由抑制作用的化学计量表示(SI＝1+k5/k4)；良好的抑制剂的值为1，因为大多数丝氨酸蛋白酶抑制蛋白分子分配为复合物形成且k5/k4接近为0。然而如果RSL环未足够快速地插入蛋白酶中，则发生分配且反应直接向裂解产物行进。
如本文所用，如本领域技术人员所熟知，催化效率或者kcat/km是蛋白酶裂解底物效率的一种测量方法，是在稳定状态条件下测量的。
如本文所用，抑制作用的二级速率常数是指蛋白酶与抑制剂相互作用的速率常数。通常蛋白酶与抑制剂如蛋白酶捕获剂如丝氨酸蛋白酶抑制蛋白的相互作用是与每种反应物、抑制剂和蛋白酶的浓度产物成比例的二级反应。由紧密结合或者不可逆的抑制剂或者蛋白酶捕获剂对于蛋白酶的抑制作用的二级速率常数是恒定的，当乘以酶浓度和抑制剂浓度时获得特定的抑制剂使酶失活的速率。每对蛋白酶捕获剂和酶的速率常数独特地反映其相互作用。作为二级反应，二级速率常数的提高意味着修饰的被选择的蛋白酶与抑制剂之间的相互作用比未修饰的蛋白酶与抑制剂之间的相互作用快。因此，二级速率常数的变化反映出反应成分即蛋白酶和/或抑制剂之间相互作用的变化。当筛选蛋白酶时，二级速率常数提高可以反映蛋白酶中希望的选择修饰。
如本文所用，酰基酶中间体是指底物与丝氨酸蛋白酶的催化中心内的必需丝氨酸(典型为Ser195，基于胰凝乳蛋白酶编号)之间催化反应第一步期间形成的共价中间体。所述反应如下进行：丝氨酸-OH攻击底物P1位置的羰基碳，组氨酸的氮接受来自丝氨酸的-OH的氢，来自羰基氧的双键的一对电子移动至所述氧原子。这样导致负电荷的四面体样中间体产生。连接底物肽键中氮与碳的键断裂。产生这个键的共价电子移动去攻击组氨酸的氢，从而破坏连接。先前从羰基氧双键移动的电子移动返回负电荷氧，再次形成键，导致共价酰基酶中间体形成。所述酰基酶中间体由水水解，导致脱酰作用并且形成裂解的底物和游离的酶。
如本文所用，蛋白酶集合是指含有至少10种不同蛋白酶和/或其蛋白酶解活性部分的集合，通常含有至少50、100、500、1000、104、105或更多个成员。所述集合典型地含有准备筛选底物特异性的蛋白酶(或其蛋白酶解活性部分)。所述集合中包括天然发生的蛋白酶(或者其蛋白酶解活性部分)和/或修饰的蛋白酶(或其蛋白酶解活性部分)。所述修饰包括沿着所述蛋白酶长度的随机突变和/或在靶定或选择区域中的修饰(即聚焦突变)。所述修饰可以是组合的，且可以包括在特定基因座或者所有基因座或者其亚集的取代所有氨基酸的所有变换。所述集合可包括全长或者较短的蛋白酶，包括仅蛋白酶结构域。所述蛋白酶可包括任何蛋白酶，例如丝氨酸蛋白酶和半胱氨酸蛋白酶。所述集合及特定集合的大小由用户确定。在其它实施方案中，所述集合含有少如2种蛋白酶。
如本文所用，“组合集合”或者“组合文库”是在其序列中具有与起始模板蛋白酶多肽序列相比独特和多样的氨基酸突变的蛋白酶多肽的集合。集合中表现的突变可以是贯穿多肽序列或者可以位于多肽序列的特定区域。所述突变可以随机产生或者可以基于结构或功能信息根据经验或者合理地设计的靶向突变。
如本文所用，“模板蛋白酶”是指具有用于诱变的氨基酸序列的蛋白酶。模板蛋白酶可以是野生型蛋白酶或者其催化活性部分的序列，或者其可以是产生另外突变的变体蛋白酶或者其催化活性部分的序列。例如，在本发明的选择方法中鉴别的突变体蛋白酶可以用作起始模板以在随后轮次的选择中进一步诱变。
如本文所用，随机突变是指不考虑突变或者对于突变没有偏好而在多肽的序列中导入一或多个氨基酸变化。随机诱变可以方便地通过本领域技术人员已知的多种技术进行，包括例如UV照射、化学方法及PCR方法(例如易错PCR)。
如本文所用，聚焦突变是指在多肽的特定区域或者特定位置的一或多个氨基酸改变。例如，可以在蛋白酶的特异性结合口袋中产生氨基酸的靶向突变。聚焦诱变可以例如使用本领域已知的标准重组技术通过定点诱变或者多位点定向诱变方法进行。
如本文所用，蛋白酶捕获剂与蛋白酶或其蛋白酶解活性部分之间的稳定复合物是指足够稳定的能从与所述蛋白酶捕获剂未形成复合物的蛋白酶(即未复合的蛋白酶)中分离出的复合物。这种复合物可以通过任何稳定的相互作用包括共价、离子和疏水性相互作用而形成，但是在反应条件下是足够稳定的以保持足够时间的复合以对复合物进行分离。丝氨酸蛋白酶抑制蛋白与裂解的蛋白酶之间的这种相互作用典型是共价键。
如本文所用，“热点”是在通过蛋白酶选择获得的对于希望的新底物序列呈现改良的活性和/或选择性的多个变体中被突变的位置。一或多个“热点”可以在蛋白酶选择期间鉴别。因此，这种位置是蛋白酶的特异性和/或选择性决定簇，并因此促成底物特异性，也可以在蛋白酶家族如丝氨酸蛋白酶家族或者特定的蛋白酶家族的一个以上成员中的相应基因座(如基于胰凝乳蛋白酶编号)作为广泛的特异性和/或选择性决定簇而发生。
如本文所用，关于底物特异性的希望的特异性是指对于预定的或预选的或者其它方式靶定的底物的裂解特异性。
如本文所用，“选择”或其语法上的变化用语是指挑选或者选择与蛋白酶捕获多肽复合的蛋白酶。因此，对于本发明而言，选择是指基于蛋白酶在稳定的复合物中与蛋白酶捕获多肽之间的结合而选出蛋白酶。通常地，选择可以通过捕获共价复合物而方便地进行，且如果需要时可以分离所述蛋白酶。例如，选择可以通过例如用预定的标记、标签或者其它可检测的部分标记所述蛋白酶捕获多肽而方便地进行，从而基于其在稳定复合物中的结合而鉴别所述蛋白酶。
如本文所用，“鉴别”及其语法上的变化用语是指识别或者了解稳定复合物中的蛋白酶。典型地，在本发明的方法中，蛋白酶是通过其在稳定复合物中与蛋白酶捕获多肽的结合而鉴别的，例如通过扩增(即在合适的宿主细胞中生长)复合物中结合的蛋白酶及随后通过DNA测序进行鉴别。
如本文所用，为了进行检测或分离而进行的标记是指使分子如蛋白酶捕获多肽与可检测的标记如荧光团或者与标签或者其它部分结合以进行纯化或者分离或者分开。可检测地标记是指对分子如蛋白酶捕获多肽标记以进行检测或分离。
如本文所用，蛋白酶或其蛋白酶解活性部分的扩增是指所述蛋白酶或其蛋白酶解活性部分的量增加，如通过分离和克隆及表达，或者在蛋白酶或其蛋白酶解活性部分在微生物上展示的情况中，将所述微生物导入合适的宿主中并且使其生长或培养以便产生更多展示的蛋白酶或其蛋白酶解活性部分。
如本文所用，与反应混合物均质是指所述反应物是液相混合物，包括溶液或悬浮液。
如本文所用，蛋白酶或其蛋白酶解活性部分的集合“基于”特定的蛋白酶是指所述集合衍生自特定的蛋白酶，例如通过随机或者定向诱变或者合理设计或者其它修饰方案产生集合。
如本文所用，通过给予t-PA治疗的疾病或病症是指本领域技术人员给予t-PA进行治疗的疾病或病症。这种病症包括但非限于纤维蛋白溶解病，如动脉血栓形成、静脉血栓形成和血栓栓塞、缺血性中风、后天凝血障碍、弥散性血管内凝血，及这些病症的前期状况，如细菌或病毒感染、牙周炎及神经系统病症。
如本文所用，由VEGFR-2介导的疾病或病症参与病理学或病因学中。这种病症包括但非限于炎症和血管生成病症，例如癌症、糖尿病视网膜病变和眼部疾病包括黄斑变性。
如本文所用，“蛋白酶”和“肽酶”可互换使用，是指催化共价肽键水解的酶。这些名称包括酶原形式及其活化的单链、双链和多链形式。为清楚起见，蛋白酶是指所有形式的蛋白酶。依赖于其活性位点的催化活性和裂解靶底物肽键的机制，蛋白酶包括例如丝氨酸蛋白酶、半胱氨酸蛋白酶、天冬氨酸蛋白酶、苏氨酸蛋白酶和金属蛋白酶。
如本文所用，酶原是指通过蛋白酶解激活的蛋白酶，包括通过成熟裂解如激活裂解和/或与其它蛋白质和/或辅因子形成复合物而激活。酶原是蛋白酶解酶的无活性前体。这种前体通常较大，但是非必需比其活性形式大。对于丝氨酸蛋白酶而言，酶原通过特异性裂解而转变为活性酶，包括催化和自身催化裂解，或者通过结合激活的辅因子，产生活性酶。因此，酶原是酶学上无活性形式的蛋白质，通过激活物的作用而将其转变为蛋白酶解酶。裂解可以通过自身催化而实现。酶原通常是无活性的，可以通过催化或者自身催化裂解所述酶原的前区(proregion)而被转变为成熟活性多肽。
如本文所用，“前区”、“前肽”或者“前序列(pro sequence)”是指被裂解以产生成熟蛋白质的区域或节段。其可包括通过掩蔽催化机构(catalyticmachinery)及因此防止催化中间体的形成(即通过在空间上封闭底物结合位点)而发挥抑制酶活性的功能的节段。前区是位于成熟的生物活性多肽的氨基末端的氨基酸序列，可以是小如几个氨基酸或者可以是多结构域结构。
如本文所用，激活序列是指酶原中为激活裂解或者成熟裂解以形成活性蛋白酶所需要的部位的氨基酸序列。激活序列的裂解可以自身催化裂解或者通过其激活配体偶催化。
激活裂解是一种成熟裂解类型，其中发生为活性所需的构象改变。这是经典的激活途径，例如对于丝氨酸蛋白酶而言，其中裂解产生新的N末端，其与催化机构的保守区域如催化残基相互作用，诱导活性所需的构象改变。激活可以导致多链形式的蛋白酶产生。在一些情况中，单链形式的蛋白酶可以呈现与单链一样的蛋白酶解活性。
如本文所用，结构域是指分子如蛋白质或者编码核酸的一部分，其在结构上和/或在功能上与所述分子的其它部分不同且是可鉴别的。
如本文所用，蛋白酶结构域是蛋白酶的催化活性部分。蛋白酶的蛋白酶结构域包括任何这些蛋白质的单链、双链和多链形式。蛋白质的蛋白酶结构域含有为其蛋白酶解活性所需的该蛋白质所有必需性质，例如其催化中心。
如本文所用，蛋白酶的催化活性部分或蛋白酶解活性部分是指蛋白酶结构域，或者其保留蛋白酶活性的任何片段或部分。重要的是，至少在体外，所述蛋白酶的单链形式及其催化结构域或者其蛋白酶解活性部分(典型为C末端截短)呈现蛋白酶活性。
如本文所用，“编码蛋白酶结构域或其催化活性部分的核酸”是指作为连续序列的仅编码所述单链蛋白酶结构域或其活性部分而不编码所述蛋白酶的其它连续部分的核酸。
如本文所用，所述基本上由蛋白酶结构域组成的多肽是指所述多肽的唯一部分是蛋白酶结构域或其催化活性部分。所述多肽可任选地且通常包括其它的非蛋白酶衍生的氨基酸序列。
如本文所用，“S1-S4”是指形成底物P1-P4残基的结合位点的氨基酸残基(见例如Schecter and Berger(1967)Biochem Biophys Res Commun 27：157-162)。每个S1-S4均含有一、二或多个残基，其可以是非连续的。这些位点从识别位点N末端至蛋白酶解位点相继编号，称作易断裂键。
如本文所用，术语“P1-P4”和“P1’-P4’”是指底物肽中的残基，其分别与S1-S4残基和S1’-S4’残基特异性相互作用，且由蛋白酶裂解。P1-P4是指位于裂解位点N末端侧的残基位置；P1’-P4’是指位于裂解位点C末端侧的残基位置。氨基酸残基从多肽底物的N末端至C末端被标记为(Pi，...，P3，P2，P1，P1′，P2′，P3′，...，Pj)。各自的结合亚位点被标记(Si，...，S3，S2，S1，S1′，S2′，S3′，...，Sj)。在P1与P1’之间催化裂解。
如本文所用，“结合口袋”是指与底物上特定氨基酸相互作用的残基。“特异性口袋”是较其它结合口袋提供更多能量的结合口袋(最重要的或者决定性的结合口袋)。典型地，结合步骤在过渡态形成之前进行，为发生催化过程所必需。S1-S4和S1’-S4’氨基酸组成底物序列结合口袋，并且通过分别与肽、多肽或者蛋白质底物的P1-P4和P1’-P4’氨基酸相互作用而促进底物识别。无论蛋白酶与底物相互作用是否是S1-S4和S1’-S4’位置中氨基酸的功能。如果任一或多个S1、S2、S3、S4、S1’、S2’、S3’和S4’亚位点中的氨基酸与底物中P1、P2、P3、P4、P1’、P2’、P3’和P4’位点的任一或多个氨基酸相互作用或者识别其，则所述蛋白酶可裂解所述底物。结合口袋使靶氨基酸与蛋白酶处于合适位置，由此实现肽键的催化和底物的裂解。例如，丝氨酸蛋白酶典型识别底物中的P4-P2’位点；其它蛋白酶可以具有超出P4-P2’的扩大的识别作用。
如本文所用，“有助于扩大底物特异性”的氨基酸是指除了特异性口袋之外活性位点裂口(cleft)中的那些残基。这些氨基酸包括蛋白酶中的S1-S4、S1’-S4’残基。
如本文所用，二级相互作用位点在活性位点裂口外面。这些位点可有助于底物识别和催化。这些氨基酸包括可导致与底物的第二和第三壳相互作用(shell interaction)的氨基酸。例如，蛋白酶结构中围绕S1-S4、S1’-S4’氨基酸的环在定位底物中的P1-P4、P1’-P4’氨基酸中起作用，从而将易断裂键记录于(registering)蛋白酶活性位点中。
如本文所用，蛋白酶的活性位点是指底物结合位点，在此发生对底物的催化。活性位点的结构和化学性质使得可以识别及结合所述底物，及随后水解和裂解底物中的易断裂键。蛋白酶的活性位点含有有助于肽裂解的催化机制的氨基酸以及有助于底物序列识别的氨基酸，例如有助于扩大底物结合特异性的氨基酸。
如本文所用，丝氨酸或半胱氨酸蛋白酶的“催化三联体”或者“活性位点残基”是指丝氨酸或半胱氨酸蛋白酶活性位点中的氨基酸组合，典型为三个氨基酸的组合，其有助于肽裂解的催化机制。通常地，催化三联体在丝氨酸蛋白酶中发现，提供活性亲核体及酸/碱催化作用。丝氨酸蛋白酶的催化三联体含有三个氨基酸，在胰凝乳蛋白酶中是Asp102、His57和Ser195。这些残基对于丝氨酸蛋白酶的催化效率是关键的。
如本文所用，“底物识别位点”或者“裂解序列”是指由蛋白酶裂解的蛋白酶活性位点识别的序列。典型地，例如对于丝氨酸蛋白酶而言，裂解序列由底物中的P1-P4和P1’-P4’的氨基酸组成，在P1位置之后发生裂解。典型地，丝氨酸蛋白酶的裂解序列的长度为6个残基，以匹配许多蛋白酶的扩大的底物特异性，但是根据蛋白酶而可以更长或更短。例如，为自身催化所需的MT-SP1的底物识别位点或裂解序列是RQARVV，其中R在P4位置，Q在P3位置，A在P2位置，R在P1位置。MT-SP1中的裂解发生在位置R后，随后是序列VVGG。
如本文所用，靶底物是指在其底物识别位点由蛋白酶特异性裂解的底物。靶底物最低程度包括组成裂解序列的氨基酸。任选地，靶底物包括含有裂解序列及任何其它氨基酸的肽。全长蛋白质、其等位基因变体、同工型或者含有由蛋白酶识别的裂解序列的任何部分是该蛋白酶的靶底物。此外，靶底物包括含有不影响蛋白酶裂解底物的其它部分的肽或蛋白质。例如，靶底物可包括与荧光部分化学连接的四氨基酸肽或者全长蛋白质。
如本文所用，裂解是指蛋白酶对肽键的破坏。蛋白酶的裂解位点基序包含易断裂键的N和C末端残基(分别为unprimed和primed sides，蛋白酶的裂解位点称作...P3-P2-P1-P1′-P2′-P3′...，裂解发生在P1与P1′残基之间)。典型地，底物的裂解是激活裂解或者抑制裂解。激活裂解是指多肽从无活性形式转变为活性形式的裂解。这种裂解包括例如酶原裂解为活性酶，和/或前生长因子(progrowth factor)裂解为活性生长因子。例如，MT-SP1可以通过在P1-P4序列RQAR裂解靶底物而自动激活。激活裂解也可以是蛋白质裂解为一或多种自身具有活性的蛋白质的裂解。例如，激活裂解在补体系统中发生，其是不可逆的蛋白酶解级联事件，其终止导致多个效应分子的形成，刺激炎症、促进抗原吞噬作用及直接溶解一些细胞。
如本文所用，抑制裂解是蛋白质裂解为一或多种非功能性降解产物。抑制裂解导致蛋白质活性的减小或降低。典型地，蛋白质活性的降低降低了涉及所述蛋白质的途径或过程。在一个实例中，任一或多种靶蛋白例如VEGFR的裂解是抑制裂解，导致靶底物的任一或多种功能或活性的伴随降低或抑制。例如，对于VEGFR的裂解，可以被抑制的活性包括但非限于配体结合、激酶活性或者血管发生活性如在体内或体外的血管发生活性。对于抑制而言，所述裂解使得活性与天然形式蛋白质的活性相比降低至少大约10％、20％、30％、40％、50％、60％、70％、80％、90％、95％、99.9％或更多。抑制裂解所需的蛋白质裂解百分比根据蛋白质而变化，但是可以通过测定蛋白质的活性而确定。
如本文所用，蛋白酶多肽是具有相应于本文所述任一候选蛋白酶或者其变体蛋白质的氨基酸序列的多肽。
如本文所用，“修饰的蛋白酶”或者“突变蛋白酶”是指在其一级序列中与野生型或模板蛋白酶相比具有一或多个修饰的蛋白酶多肽(蛋白质)。所述一或多个突变可以是一或多个氨基酸置换(取代)、插入、缺失及其任意组合。修饰的蛋白酶多肽包括具有1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20或更多个修饰位置的那些蛋白酶多肽。修饰的蛋白酶可以是全长蛋白酶，或者可以是修饰的全长蛋白酶的催化活性部分，只要所述修饰的蛋白酶是蛋白酶解活性的。通常地，这些突变改变了野生型或者模板蛋白酶裂解任一或多种希望的或预定的靶底物的特异性和活性。除了在改变蛋白酶的底物特异性的区域中含有修饰之外，修饰的蛋白酶在对于蛋白酶的底物特异性非必需的区域中也可允许具有其它修饰。因此，修饰的蛋白酶与相应的野生型或者支架蛋白酶的氨基酸序列典型具有60％、70％、80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多的序列相同性。修饰的全长蛋白酶或者修饰的蛋白酶的其催化活性部分可包括是融合蛋白的蛋白酶，只要所述融合蛋白自身不改变蛋白酶的底物特异性。
如本文所用，胰凝乳蛋白酶编号是指SEQ ID NO：391所示成熟胰凝乳蛋白酶多肽的氨基酸编号。胰凝乳蛋白酶家族蛋白酶(即u-PA、t-PA、MT-SP1及其它成员)包括蛋白酶结构域的对比可以用胰凝乳蛋白酶进行。在这种情况中，相应于胰凝乳蛋白酶的氨基酸的蛋白酶的氨基酸以胰凝乳蛋白酶氨基酸的编号示出。相应位置可以由本领域技术人员使用人工对比方式或者使用多种可利用的对比程序(例如BLASTP)通过这种对比而确定。相应位置也可以基于结构对比，例如通过使用计算机模拟的蛋白酶结构对比而确定。相应于揭示的序列中氨基酸的多肽的氨基酸是指使用标准对比算法如GAP算法对比所述多肽与所揭示的序列以使得相同性或同源性最大化(其中对比保守氨基酸)而鉴别的氨基酸。例如，在对比u-PA与成熟胰凝乳蛋白酶多肽时，SEQ ID NO：191所示u-PA的前体序列中氨基酸C168与成熟胰凝乳蛋白酶多肽的氨基酸C1对比在一起。因此，u-PA中的氨基酸C168基于胰凝乳蛋白酶编号也是C1。使用这种胰凝乳蛋白酶编号标准，SEQ ID NO：191所示前体u-PA序列中氨基酸L244与基于胰凝乳蛋白酶编号的L73相同，氨基酸I260与基于胰凝乳蛋白酶编号的I89相同。在另一实例中，在对比MT-SP1丝氨酸蛋白酶结构域(相应于SEQ ID NO：253的第615-855位氨基酸)与成熟胰凝乳蛋白酶时，在MT-SP1第615位的V以胰凝乳蛋白酶编号V16示出。随后的氨基酸相应编号。因此，在全长MT-SP1(SEQ ID NO：253)第708位的氨基酸F基于胰凝乳蛋白酶编号相应于F99。在残基存在于蛋白酶中而在胰凝乳蛋白酶中不存在的情况中，该氨基酸残基以字母表示。例如，是胰凝乳蛋白酶中是具有氨基酸60(基于胰凝乳蛋白酶编号)的环的一部分但是与胰凝乳蛋白酶相比插入在MT-SP1序列中的残基，例如称作Asp60b或Arg60c。
如本文所用，对于靶底物的特异性是指与另一底物(称作非靶底物)相比蛋白酶优先裂解靶底物。特异性以二级速率常数或特异性常数反映(kcat/Km)，这是蛋白酶与其底物的亲和性及该酶的效率的测量数据。
如本文所用，对于裂解的特异性常数是(kcat/Km)，其中Km是Michaelis-Menton常数(在一半Vmax的[S])，Kcat是Vmax/[ET]，其中ET是最终的酶浓度。参数kcat、Km和kcat/Km可以通过将底物浓度的倒数对底物裂解速度的倒数作图并拟合进Lineweaver-Burk方程(1/速度＝(Km/Vmax)(1/[S])+1Vmax，其中Vmax＝[ET]kcat)而计算。可以使用在存在不同浓度底物的条件下确定裂解速度随着时间而增加的任何方法计算特异性常数。例如，将底物与荧光部分连接，其在由蛋白酶裂解时释放。通过确定在不同酶浓度下的裂解速度，可以确定特定蛋白酶的kcat。特异性常数可以用于使用本领域的标准方法例如使用位置扫描组合文库(positional scanning combinatoriallibrary，PS-SCL)确定蛋白酶的任一或多个S1-S4口袋中的氨基酸相对于底物中伴随的P1-P4氨基酸的位点特异性偏爱。此外，特异性常数也可以用于确定蛋白酶对于一个靶底物相对于另一个靶底物的偏爱。
如本文所用，底物特异性比率是特异性常数的比率，可用于对比两或多个蛋白酶的特异性或者蛋白酶对于两个以上底物的特异性。例如，蛋白酶对于竞争底物的底物特异性或者竞争蛋白酶对于底物的底物特异性可以通过对比kcat/Km而进行比较。例如，对于靶底物具有2×106M-1sec-1特异性常数或者对于非靶底物具有2×104M-1sec-1特异性常数的蛋白酶对于所述靶底物更具特异性。使用上述特异性常数，所述蛋白酶对于靶蛋白酶具有100的底物特异性比率。
如本文所用，对于靶底物的偏爱可以底物特异性比率表示。反映偏爱性的特定比率值是底物与所研究的蛋白酶之间的函数。大于1的底物特异性比率表示对于靶底物的偏爱，小于1的底物特异性比率表示对于非靶底物的偏爱。通常地，至少或者大约为1的比率反映出被认为是候选治疗剂的蛋白酶的充分差异。
如本文所用，改变的特异性是指与起始野生型或模板蛋白酶相比修饰的或选择的蛋白酶的底物特异性变化。通常，特异性变化是与模板蛋白酶的野生型底物(在此称作非靶底物)相比修饰的蛋白酶对于靶底物的偏爱性变化。典型地，与靶蛋白酶的底物特异性相比，本发明提供的修饰的蛋白酶或者选择的蛋白酶对于靶蛋白质的任一或多种预定或希望的裂解序列呈现增加的底物特异性。例如，对于靶底物相对于与非靶底物的底物特异性比率为100的修饰的蛋白酶或者选择的蛋白酶与底物特异性比率为10的支架蛋白酶相比呈现出特异性增加10倍。在另一实例中，底物特异性比率为1相对于底物特异性比率为0.1的修饰的蛋白酶呈现出底物特异性增加10倍。为了呈现出与模板蛋白酶相比增加的特异性，修饰的蛋白酶对于任一或多种预定靶底物具有1.5倍、2倍、5倍、10倍、50倍、100倍、200倍、300倍、400倍、500倍或更高的底物特异性。
如本文所用，当用于描述蛋白酶从竞争底物混合物中选择和裂解一种靶底物的能力时，“选择性”可以与特异性互换使用。可以通过例如对比蛋白酶裂解靶底物的特异性常数而确定与任一或多种靶底物相比，蛋白酶对于一种靶底物增加的选择性。例如，如果蛋白酶对于靶底物的裂解特异性常数为2×106M-1sec-1而对于其它任一其他底物的裂解特异性常数为2×104M-1sec-1，则该蛋白酶对于前者靶底物更具选择性。
如本文所用，活性是指多肽或其与全长(完整)蛋白质相关联的一部分的一或多种功能活性。功能活性包括但非限于生物学活性、催化或酶活性、抗原性(结合或者与多肽竞争结合抗多肽抗体的能力)、免疫原性、形成多聚体的能力及特异性结合所述多肽的受体或配体的能力。
如本文所用，催化活性或者裂解活性是指在检测选择的底物的蛋白酶解作用的体外蛋白酶解测定中评价的蛋白酶的活性。裂解活性可以通过评价蛋白酶的催化效率而测量。
如本文所用，对于靶底物的活性是指裂解活性和/或功能活性，或者反映蛋白酶对于靶底物的活性的其它量度。裂解活性可以通过评价蛋白酶的催化效率而测量。对于本发明而言，如果与不存在蛋白酶的条件相比，蛋白酶呈现出更高的对靶底物的蛋白酶解或者裂解作用和/或调节(即激活或抑制)靶底物蛋白的功能活性，则表示活性增加。
如本文所用，丝氨酸蛋白酶或者丝氨酸内肽酶是指一类肽酶，其特征在于在酶的活性中心存在丝氨酸残基。丝氨酸蛋白酶参与机体内的许多功能，包括血液凝固、炎症以及在原核生物和真核生物中的消化酶。由“丝氨酸蛋白酶”所致的裂解机制是基于丝氨酸对于靶定肽键的亲核攻击。与天冬氨酸相关的半胱氨酸、苏氨酸或水分子或金属也可以发挥这个作用。丝氨酸、组氨酸和天冬氨酸的排列的侧链形成大多数丝氨酸蛋白酶共有的催化三联体。丝氨酸蛋白酶的活性位点在多肽底物结合之处是裂口形状。丝氨酸蛋白酶的例子包括SEQ ID NO：433所示尿纤溶酶原激活物(u-PA)及SEQ ID NO：253所示MT-SP1，及其催化活性部分，例如SEQ ID NO：505所示MT-SP1蛋白酶结构域(也称作B-链)。
如本文所用，人蛋白质是由人基因组中存在的核酸分子如DNA编码的蛋白质，包括其所有等位基因变体和保守变体。如果修饰基于人蛋白质的野生型或者主要序列，则蛋白质的变体或修饰是人蛋白质。
如本文所用，天然发生的α-氨基酸残基是在自然界发现的那20种α-氨基酸残基，其通过负载tRNA分子由人体中关联mRNA密码子的特异性识别而掺入蛋白质中。
如本文所用，非天然发生的氨基酸是指非遗传编码的氨基酸。
如本文所用，核酸包括DNA、RNA及其类似物，包括肽核酸(PNA)及其混合物。核酸可以是单链或双链的。当用于描述探针或引物时，包括单链分子，其任选被标记，如用可检测的标记如荧光或放射性标记进行标记。这种分子典型是这样的长度，以便其靶位用于探查或引发文库时在统计学上是唯一的或者是低拷贝数的(典型小于5，通常小于3)。通常探针或引物含有与感兴趣的基因互补或相同的序列的至少14、16或30个连续核苷酸。探针和引物的长度可以是10、20、30、50、100或更多个核酸。
如本文所用，肽是指长度为2-40个氨基酸的多肽。
如本文所用，在本发明提供的各个氨基酸序列中出现的氨基酸根据其已知的三字母或单字母缩写的方式表示(表1)。在各个核酸片段中出现的核苷酸以本领域常用的标准单字母命名法命名。
如本文所用，“氨基酸”是含有氨基基团和羧酸基团的有机化合物。多肽含有两或多个氨基酸。对于本发明而言，氨基酸包括20种天然发生的氨基酸、非天然氨基酸及氨基酸类似物(即其中α-碳具有侧链的氨基酸)。
如本文所用，“氨基酸残基”是指多肽在其肽键部位被化学消化(水解)时形成的氨基酸。本文所述的氨基酸残基推定为L异构体形式。也如此命名的D异构体形式的残基可以由任何L-氨基酸残基取代，只要多肽保留希望的功能性质。NH2是指在多肽的氨基末端存在的游离氨基基团。COOH是指在多肽的羧基末端存在的游离羧基基团。与J.Biol.Chem.，243：3552-3559(1969)及采用的37C.F.R..§§1.821-1.822所述标准多肽命名法一致，表1示出了氨基酸残基的缩写。
表1-对应表
应注意本文示出的所有氨基酸残基序列均是从左至右方向是氨基末端至羧基末端常规方向。另外，短语“氨基酸残基”广泛是指包括对应表(表1)中列出的氨基酸及修饰的和不常见的氨基酸，如并入作参考的37C.F.R.§§1.821-1.822中提及的那些。此外，应注意在氨基酸残基序列的起始处或末端的破折号“-”表示与一或多个氨基酸残基的进一步序列的肽键，与氨基末端基团如NH2或者与羧基末端基团如COOH的肽键。
如本文所用，“天然发生的氨基酸”是指在多肽中出现的20种L-氨基酸。
如本文所用，“非天然氨基酸”是指这样的有机化合物，其具有与天然氨基酸相似的结构，但是在结构上已经被修饰为模拟天然氨基酸的结构和反应性。非天然发生的氨基酸因此包括例如除了所述20种天然发生的氨基酸之外的氨基酸或氨基酸类似物，但非限于氨基酸的D-立体异构体(D-isostereomer)。非天然氨基酸的例子在本文中描述，且为本领域技术人员所已知。
如本文所用，等动力混合物是其中氨基酸的摩尔比率已经基于其报道的反应速率而调节的混合物(见例如Ostresh et al.，(1994)Biopolymers34：1681)。
如本文所用，DNA构建体是单链或双链、线性或环形DNA分子，含有以天然未发现的方式组合和并列的DNA节段。DNA构建体以人工操纵的结果而存在，包括操纵的分子的克隆及其它拷贝。
如本文所用，DNA节段是具有特定属性的较大DNA分子的一部分。例如，编码特定多肽的DNA节段是较长的DNA分子的一部分，如质粒或质粒片段，当从5’至3’方向读取时，其编码特定多肽的氨基酸序列。
如本文所用，术语直向同源物是指得自一个物种的是功能对应物的多肽或蛋白质，或者来自不同物种的多肽或蛋白质。直向同源物的序列差异是物种形成的结果。
如本文所用，术语多核苷酸是指从5’至3’末端读取的脱氧核糖核苷酸或者核糖核苷酸碱基的单链或双链聚合物。多核苷酸包括RNA和DNA，可以分离自天然来源、在体外合成或者从天然分子与合成分子的组合中制备。多核苷酸分子的长度在本文以核苷酸(缩写为nt)或碱基对(缩写为bp)示出。术语核苷酸根据上下文用于描述单链和双链分子。当该术语用于双链分子时，其用于表示全长分子，且应理解为与术语碱基对相同。本领域技术人员意识到双链多核苷酸的两个链在长度上可以略微不同，其末端可以是交错的；因此双链多核苷酸分子内的所有核苷酸可以不都是配对的。这种不配对的末端通常不超过20个核苷酸长度。
如本文所用，两个蛋白质或核酸之间的“相似性”是指蛋白质的氨基酸序列或者核酸的核苷酸序列之间的关联性。相似性可以基于残基序列和其中包含的残基的相同性和/或同源性程度。评价蛋白质或核酸之间相似性程度的方法为本领域技术人员所已知。例如，在评价序列相似性的一个方法中，将两个氨基酸或核苷酸序列以在序列之间产生最大水平相同性的方式对比。“相同性”是指氨基酸或者核苷酸序列无差异的程度。氨基酸序列的对比及在一定程度上核苷酸序列的对比也可以考虑到氨基酸(或者核苷酸)中的保守差异和/或频繁取代。保守差异是保持所包含残基的物理-化学性质的那些差异。对比可以是整体对比(对全长序列进行对比，包括所有残基)或者局部对比(仅包括最相似区域的序列部分的对比)。
“相同性”具有本领域公认的含义，可以使用公开的技术计算(见例如Computational Molecular Biology，Lesk，A.M.，ed.，Oxford University Press，New York，1988；Biocomputing：Informatics and Genome Projects，Smith，D.W.，ed.，Academic Press，New York，1993；Computer Analysis of Sequence Data，Part I，Griffin，A.M.，and Griffin，H.G.，eds.，Humana Press，New Jersey，1994；Sequence Analysis in Molecular Biology，von Heinje，G.，Academic Press，1987；及Sequence Analysis Primer，Gribskov，M.and Devereux，J.，eds.，M StocktonPress，New York，1991)。存在测量两个多核苷酸或多肽之间相同性的许多方法，术语“相同性”为本领域技术人员所熟知(Carillo，H.& Lipton，D.，SIAMJ Applied Math 48：1073(1988))。
如本文所用，同源(关于核酸和/或氨基酸序列)是指大约高于或等于25％序列同源性，典型高于或等于25％、40％、50％、60％、70％、80％、85％、90％或95％序列同源性；如果需要，可以指明精确的百分比。对于本发明而言，除非特别指出，则术语“同源性”和“相同性”通常可互换使用。通常地，为了确定同源性或相同性百分比，对比序列以便获得最高级别的匹配(见例如Computational Molecular Biology，Lesk，A.M.，ed.，OxfordUniversity Press，New York，1988；Biocomputing：Informatics and GenomeProjects，Smith，D.W.，ed.，Academic Press，New York，1993；ComputerAnalysis of Sequence Data，Part I，Griffin，A.M.，and Griffin，H.G.，eds.，Humana Press，New Jersey，1994；Sequence Analysis in Molecular Biology，vonHeinje，G.，Academic Press，1987；及Sequence Analysis Primer，Gribskov，M.and Devereux，J.，eds.，M Stockton Press，New York，1991；Carillo et al.(1988)SIAM J Applied Math 48：1073)。通过序列同源性，保守氨基酸的数目通过标准对比算法程序确定，可以使用每个供应商确定的默认缺口罚分。基本上同源的核酸分子典型在中等严格条件或者在高度严格条件下沿着感兴趣的核酸的长度杂交。本发明还涉及含有简并密码子代替杂交核酸分子中的密码子的核酸分子。
任两个分子是否具有至少60％、70％、80％、85％、90％、95％、96％、97％、98％或99％“相同”或“同源”的核苷酸序列或氨基酸序列可以使用已知的计算机算法如FASTA程序利用例如默认参数确定，如Pearson et al.(1988)Proc.Natl.Acad.Sci.USA 85：2444所述(其它程序包括GCG程序包(Devereux，J.，et al.，Nucleic Acids Research 12(I)：387(1984))、BLASTP、BLASTN、FASTA(Atschul，S.F.，et al，JMolec Biol 215：403(1990))；Guide toHuge Computers，Martin J.Bishop，ed.，Academic Press，San Diego，1994及Carillo et al.(1988)SIAM J Applied Math 48：1073)。例如，National Center forBiotechnology Information数据库的BLAST可用于确定相同性。其它可商购或可公开获得的程序包括DNAStar″MegAlign″程序(Madison，WI)和theUniversity of Wisconsin Genetics Computer Group(UWG)″Gap″程序(MadisonWI)。蛋白质和/或核酸分子的同源性或者相同性百分比可以例如通过使用GAP计算机程序对比序列信息而确定(例如Needleman et al.(1970)J.Mol.Biol.48：443，由Smith and Waterman((1981)Adv.Appl.Math.2：482修订)。简而言之，GAP程序将相似性规定为相似的对比符号(即核苷酸或氨基酸)的数目除以两个序列中较短序列的符号总数。GAP程序的默认参数可包括：(1)一元比较矩阵(含有相同性值为1，非相同性值为0)及Gribskov et al.(1986)Nucl.Acids Res.14：6745的加权比较矩阵，如Schwartz and Dayhoff，eds.，ATLAS OF PROTEIN SEQUENCE AND STR UCTURE，NationalBiomedical Research Foundation，pp.353-358(1979)所述；(2)每个缺口的3.0罚分及每个缺口中每个符号的额外0.10罚分；及(3)末端缺口无罚分。
因此，如本文所用，术语“相同性”或者“同源性”表示测试多肽或多核苷酸与参考多肽或多核苷酸之间的对比。如本文所用，术语至少“90％相同”是指相对于参考核酸或多肽的氨基酸序列从90至99.99的相同性百分比。在90％或者更高水平的相同性是指假定对比长度为100个氨基酸的测试多肽和参考多肽。在测试多肽中不超过10％的(即100分之10)氨基酸与参考多肽的氨基酸不同。在测试多核苷酸与参考多核苷酸之间可以进行相似对比。这种差异可以表示随机分布于多肽全长的点突变，或者其可以聚集在不同长度的一或多个位置直至最大容许值，例如10/100氨基酸差异(大约90％相同性)。差异定义为核酸或者氨基酸取代、插入或者缺失。在大约85-90％以上的同源性或相同性水平，结果应与程序和缺口参数设置无关，这种高水平相同性可以易于通过人工对比而不用依靠软件即可评定。
如本文所用，对比的序列是指使用同源性(相似性和/或相同性)对比核苷酸或氨基酸序列中的相应位置。典型地，对比50％或更高相同性的相关的两或多个序列。对比的序列组是指在相应位置对比的两或多个序列，且可以包括衍生自RNA的对比序列，例如EST及与基因组DNA序列对比的其它cDNA。
如本文所用，“引物”是指可作为模板指导的DNA合成的起始点的核酸分子，所述合成是在适当条件下(例如存在四种不同的核苷三磷酸和聚合剂如DNA聚合酶、RNA聚合酶或者逆转录酶)、在合适的缓冲液中及在合适的温度下进行。本领域技术人员意识到某些核酸分子可以作为“探针”和“引物”。然而，引物具有3’羟基基团以进行延伸。引物可用于多种方法中，包括例如聚合酶链反应(PCR)、逆转录酶(RT)-PCR、RNA PCR、LCR、多重PCR、panhandle PCR、捕获PCR、表达PCR、3′和5′RACE、原位PCR、连接介导的PCR及其它扩增方法。
如本文所用，“引物对”是指一组引物，其包括与被扩增(例如通过PCR扩增)的序列的5’末端杂交的5’(上游)引物及与被扩增的序列的3’末端的互补序列杂交的3’(下游)引物。
如本文所用，“特异性杂交”是指通过互补碱基配对，核酸分子(例如寡核苷酸)与靶核酸分子退火。本领域技术人员熟知影响特异性杂交的体外和体内参数，例如特定分子的长度和组成。与体外杂交特别相关的参数进一步包括退火和洗涤温度、缓冲液组成和盐浓度。在高度严格条件下，除去非特异性结合的核酸分子的洗涤条件例如是0.1 x SSPE、0.1％SDS、65℃，在中等条件下是0.2 x SSPE、0.1％SDS、50℃。等价的严格条件为本领域所已知。技术人员可容易地调节这些参数以实现核酸分子与适于特定用途的靶核酸分子的特异性杂交。
如本文所用，与产物基本上相同是指足够的相似，由此感兴趣的性质足以未加改变以便所述基本上相同的产物可用于代替所述产物。
如本文所用，也应理解术语“基本上相同”或者“相似”根据上下文而有所不同，这种用法为相关领域技术人员所了解。
如本文所用，等位基因变体或者等位基因变异是指占据相同染色体基因座的基因的两或多种变化形式任一。等位基因变异通过突变天然产生，且可以导致群体内的表型多态性。基因突变可以是沉默突变(编码的多肽中无变化)或者可以编码具有改变的氨基酸序列的多肽。术语“等位基因变体”在本文也用于描述由基因的等位基因变体编码的蛋白质。典型地，基因的所述参考形式编码来自群体或者来自物种的一个参考成员中野生型形式和/或优势形式多肽。典型地，等位基因变体包括物种之间或者物种之中的变体，与相同物种的野生型和/或优势形式典型具有至少80％、90％或更高的氨基酸相同性；相同性的程度取决于基因及对比是种间还是种内对比。通常，种内等位基因变体与野生型和/或优势形式具有至少大约80％、85％、90％或95％或者更高的相同性，包括与多肽的野生型和/或优势形式具有96％、97％、98％、99％或者更高的相同性。本文提及的等位基因变体通常是指相同物种的成员之中的变异蛋白质。
如本文所用，“等位基因”与“等位基因变体”在本文可互换使用，是指基因或其一部分的替换形式。等位基因占据同源染色体上的相同基因座或位置。当对象具有基因的两个相同的等位基因时，称所述对象是该基因或等位基因纯合的。当对象具有基因的两个不同的等位基因时，称所述对象是该基因杂合的。特定基因的等位基因彼此可以有一个核苷酸或者几个核苷酸的不同，且可以包括核苷酸的取代、缺失和插入。基因的等位基因可以是含有突变的基因形式。
如本文所用，物种变体是指不同物种中的多肽变体，包括不同的哺乳动物物种如小鼠和人。
如本文所用，剪接变体是指通过对基因组DNA的一级转录物进行不同加工，产生一种以上类型的mRNA而产生的变体。
如本文所用，修饰是指多肽的氨基酸序列或者核酸分子的核苷酸序列中的修饰，分别包括氨基酸和核苷酸的缺失、插入和置换。修饰多肽的方法为本领域技术人员所熟知，例如使用重组DNA方法学进行。
如本文所用，肽模拟物(peptidomimetic)是模拟特定肽的生物学活性形式的构象和某些立体化学特征的化合物。通常，肽模拟物被设计为模拟化合物的某些希望的性质而不模拟不希望的性质，例如导致生物学活性构象丧失和键破坏的柔性。通过用生物等构物(bioisostere)置换导致不希望的性质的某些基团或键，可以从生物学活性化合物制备肽模拟物。生物等构物为本领域技术人员所已知。例如，亚甲基生物等构物CH2S已经在脑啡肽类似物中用作酰胺置换物(见例如Spatola(1983)pp.267-357in Chemistryand Biochemistry of Amino Acids，Peptides，and Proteins，Weinstein，Ed.volume 7，Marcel Dekker，New York)。可以口服的吗啡是内啡肽的肽模拟物化合物。对于本发明而言，环肽包括在肽模拟物中，是其中一或多个肽键由模拟物置换的多肽。
如本文所用，一种多肽包含特定百分比的参考多肽的氨基酸是指在多肽与参考多肽之间共有的连续相同氨基酸的比例。例如，包含具有SEQ IDNO：XX所示氨基酸序列(其列举了147个氨基酸)的参考多肽中70％氨基酸的同工型是指所述参考多肽含有SEQ ID NO：XX所示氨基酸序列的至少103个连续氨基酸。
如本文所用，术语启动子是指含有用于RNA聚合酶结合及转录起始的DNA序列的基因部分。启动子序列通常但非总是在基因的5’非编码区中发现。
如本文所用，分离的或纯化的多肽或蛋白质或其生物学活性部分基本上不含细胞材料或者来自从中衍生所述蛋白质的细胞或组织中的其它污染蛋白质，或者当其是经化学合成的时，基本上不含化学前体或其它化学物。如果通过标准分析方法确定制备物看起来没有易于可检测的杂质，则可以确定所述制备物基本上是不含污染物的，所述方法如薄层层析法(TLC)、凝胶电泳法和高效液相层析法(HPLC)，本领域技术人员使用这些方法评定这种纯度，或者所述制备物是足够纯的由此进一步的纯化将不会可检测地改变所述物质的物理和化学性质，如酶和生物学活性。纯化化合物以产生基本上化学纯的化合物的方法为本领域技术人员所已知。基本上化学纯的化合物可以是立体异构体的混合物。在这种情况中，进一步纯化可增加化合物的比活性。
术语“基本上不含细胞材料”包括这样的蛋白质制备物，其中蛋白质自从中分离或者重组产生蛋白质的细胞的细胞成分中分离出。在一个实施方案中，术语基本上不含细胞材料包括这样的蛋白酶蛋白质制备物，其具有低于大约30％(干重)的非蛋白酶蛋白质(在本文也称作污染蛋白质)，通常低于大约20％的非蛋白酶蛋白质或者10％的非蛋白酶蛋白质或者低于大约5％的非蛋白酶蛋白质。当所述蛋白酶蛋白质或其活性部分是重组产生的时，其也基本上不含培养基，即培养基低于大约或者为所述蛋白酶蛋白质制备物体积的20％、10％或5％。
如本文所用，术语基本上不含化学前体或者其它化学物包括这样的蛋白酶蛋白质制备物，其中所述蛋白质从参与蛋白质合成的化学前体或者其它化学物中分离出。该术语包括这样的蛋白酶蛋白质制备物，其具有低于大约30％(干重)、20％、10％、5％或更少的化合物前体或非蛋白酶化学物或成分。
如本文所用，术语“合成的”，例如合成的核酸分子或者合成的基因或者合成的肽是指核酸分子或者多肽分子是通过重组方法和/或通过化学合成方法产生的。
如本文所用，通过使用重组DNA方法重组产生是指使用分子生物学领域熟知的方法表达由克隆的DNA编码的蛋白质。
如本文所用，载体(或质粒)是指用于将异源核酸导入细胞中以进行表达或复制的不连续元件。载体典型地保持为附加体，但是可以设计为实现基因或其一部分整合进基因组染色体中。本发明还涉及作为人工染色体的载体，例如酵母人工染色体和哺乳动物人工染色体。这种载体的选择和应用为本领域技术人员所熟知。
如本文所用，表达载体包括能表达DNA的载体，所述DNA与能实现这种DNA片段表达的调节序列如启动子区域可操纵地连接。这种额外的节段可包括启动子和终止子序列，及任选可包括一或多个复制起点、一或多个可选择的标记、增强子、聚腺苷酸化信号等。表达载体通常衍生自质粒或病毒DNA，或者可以含有这两种元件。因此，表达载体是指重组DNA或RNA构建体，如质粒、噬菌体、重组病毒或者其它载体，在导入合适的宿主细胞中时导致克隆的DNA表达。合适的表达载体为本领域技术人员所熟知，包括在真核细胞和/或原核细胞中可以复制的那些载体，及保持为附加体的那些载体或者整合进宿主细胞基因组中的那些载体。
如本文所用，载体还包括“病毒载体”。病毒载体是工程化的病毒，其与外源基因可操纵地连接以(作为运载体或穿梭工具)将所述外源基因转移进细胞中。
如本文所用，腺病毒是指在人体内导致结膜炎和上呼吸道感染的含有DNA的任何病毒。如本文所用，裸DNA是指无组蛋白的DNA，其可以用于疫苗和基因治疗。裸DNA是遗传材料，其在称作转化的基因转移过程中在细胞与细胞中传递。在转化中，纯化的或裸DNA由受体细胞摄取，其将为受体细胞提供新的特性或表型。
如本文所用，可操纵地连接当用于描述DNA节段时是指所述节段的排列使得其功能与指定目的一致，例如转录在启动子中起始且从编码节段行进至终止子。
如本文所用，蛋白质结合序列是指能特异性结合其它蛋白质或肽序列、通常是一组蛋白质或肽序列或者特定的蛋白质或肽序列的蛋白质或肽序列。
如本文所用，表位标记是指相应于表位的一段较短的氨基酸残基序列，便于随后对表位标记的蛋白质或肽进行生物化学和免疫学分析。表位标记是通过将表位标记序列加入在合适的表达载体中的蛋白酶编码序列中而实现。表位标记的蛋白质可以通过使用针对所述标记的高度特异性抗体进行亲和纯化。
如本文所用，金属结合序列是指能特异性结合金属离子、通常为一组金属离子或者特定的金属离子的蛋白质或肽序列。
如本文所用，术语评定是指包括定量和定性确定，获得样品中存在的蛋白质或其结构域活性的绝对值，也可以获得关于活性水平的指数、比率、百分比、目测值或者其它数值。评定可以是直接或间接评定，且实际检测的化学种类不需要是自身蛋白酶解产物，而是可例如是其衍生物或者一些进一步的物质。例如，通过SDS-PAGE及用考马斯蓝染色蛋白质检测补体蛋白的裂解产物。
如本文所用，生物学活性是指化合物的体内活性或者在体内给予化合物、组合物或者其它混合物时产生的生理反应。因此，生物学活性包含这种化合物、组合物和混合物的治疗效果和药物活性。生物学活性可以在设计为测试或使用这种活性的体外系统中观测。因此，对于本发明而言，蛋白酶的生物学活性是其催化活性，其中多肽被水解。
如本文所用，当用于描述核酸的两个序列时，术语等价是指所述两个序列编码相同的氨基酸序列或者等价的蛋白质。当等价用于描述两个蛋白质或肽时，是指这两个蛋白质或肽具有基本相同的氨基酸序列，仅具有基本不改变所述蛋白质或肽的活性或功能的氨基酸取代。当等价用于描述性质时，所述性质不需要以相同程度存在(例如两个肽可以表现为不同速率的相同类型酶活性)，但是所述活性通常基本相同。当用于描述两个核苷酸序列时，互补是指两个核苷酸序列能杂交，在相对的核苷酸之间典型具有低于25％、15％或5％的错配。如果需要，指明互补百分比。典型地，选择在高度严格条件下杂交的两个分子。
如本文所用，调节蛋白质活性或者基因或核酸表达的物质降低或者增加或者另外改变蛋白质的活性，或者以一些方式上调或下调或者另外改变核酸在细胞中的表达。
如本文所用，药物作用或者治疗作用是指在给予用于治疗疾病或病症或者改善其症状的物质时观测到的作用。
如本文所用，“调节”或“改变”是指分子例如蛋白质的活性的改变。活性例如包括但非限于生物学活性，如信号转导。调节可包括活性增加(即上调或激动剂活性)、活性降低(即下调或抑制)或者活性的任何其它改变(如周期性、频率、持续时间、动力学或者其它参数的改变)。调节可以根据情况而定，典型地，调节是与指定状态相对比，例如与野生型蛋白质、组成型状态的蛋白质或者在指定细胞类型或条件中表达的蛋白质相对比。
如本文所用，抑制是指相对于未被抑制的活性，活性降低。
如本文所用，组合物是指任何混合物。其可以是溶液、悬浮液、液体、粉末、糊剂、水溶液、非水溶液或者其任意组合。
如本文所用，组合是指两或多个项目(items)之间或之中的任何结合。所述组合可以是两或多个单独项目，如两个组合物或两个集合，可以是其混合物，如两或多个项目的单一混合物，或者其任何变体。组合的元件通常是功能关联或相关的。试剂盒是包装的组合，其任选包括使用所述组合或其元件的说明书。
如本文所用，“疾病或失调”是指生物体由于一些原因或条件而引起的病理学病症，所述原因和条件包括但非限于感染、后天获得的病症、遗传病症，及特征在于可鉴别的症状。本发明感兴趣的疾病或失调是涉及补体激活的那些，包括由补体激活介导的那些及其中补体激活在病因学和病理学中起作用的那些。疾病和失调也包括由于蛋白质的缺乏而引起的那些，如免疫缺陷，本发明感兴趣的疾病和失调是其中由于补体蛋白的缺陷而不发生补体激活的那些失调。
如本文所用，“治疗”患有疾病或病症的对象是指在治疗后所述对象的症状部分或全部减轻，或者保持稳定状态。因此，治疗包含预防、治疗和/或治愈。预防是指预防潜在的疾病和/或预防症状恶化或者疾病进展。治疗还包含本文提供的修饰的干扰素和组合物的药物学应用。
如本文所用，治疗剂、治疗方案、放射防护剂或者化学疗法是指常规的药物和药物治疗，包括疫苗，这些为本领域技术人员所已知。放射疗法为本领域所熟知。
如本文所用，治疗是指其中病症、失调或疾病的症状或者其它指征得以改善或者另外得以有益改变的任何方式。
如本文所用，治疗作用是指对对象进行治疗后获得的改变、典型为改善或者减轻疾病或病症的症状或者治愈疾病或病症的作用。治疗有效量是指在给予对象之后产生治疗作用的组合物、分子或者化合物的量。
如本文所用，术语“对象”是指动物，包括哺乳动物，如人类。
如本文所用，患者是指人对象。
如本文所用，通过例如给予药物组合物或者其它治疗方法进行处理而改善特定疾病或失调的症状是指由于给予所述组合物或者治疗方法引起的或者与之相关的症状的任何减轻，无论是永久或暂时、长期或短暂的。
如本文所用，预防是指降低发生疾病或病症的风险的方法。
如本文所用，有效量是预防、治愈、改善、阻止或部分阻止疾病或失调的症状必需的治疗剂的量。
如本文所用，给予蛋白酶如修饰的蛋白酶是指将所述蛋白酶与其底物接触的任何方法。可以在体内或离体或在体外给予。例如，对于离体给予，将体液如血液从对象体内取出，并且在机体之外与修饰的非补体蛋白酶接触。对于体内给予，可以将修饰的蛋白酶导入机体中，如通过局部、表面、全身和/或其它导入途径。体外给予包含如细胞培养方法等方法。
如本文所用，如本领域技术人员所已知，单位剂量形式是指适于人和动物对象且单独包装的物理分立单位。
如本文所用，单一剂量制剂是指直接给予的制剂。
如本文所用，“制造品”是生产和销售的产品。如本说明书中所用，该术语包含包装物品中包含的修饰的蛋白酶多肽及核酸。
如本文所用，流体是指可以流动的任何组合物。因此，流体包含半固体、糊剂、溶液、水溶液混合物、凝胶、洗剂、乳液及其它这种组合物形式的组合物。
如本文所用，“试剂盒”是指本发明提供的修饰的蛋白酶多肽或者核酸分子与另一项目的组合，所述另一项目用于包括但非限于给予、诊断和评定生物学活性或性质的项目。试剂盒典型包括使用说明书。
如本文所用，细胞提取物或者溶解产物是指从溶解或破坏的细胞中制备的制备物或级分。
如本文所用，动物包括任何动物，例如但非限于灵长类动物包括人、大猩猩和猴子；啮齿类动物如小鼠和大鼠；禽类如鸡；反刍动物如山羊、牛、鹿、绵羊；猪类如猪，及其它动物。非人动物是除了人之外的动物。本发明提供的蛋白酶来自任何来源，动物、植物、原核生物和真菌。大多数蛋白酶源于动物，包括源于哺乳动物。
如本文所用，对照是指与测试样品基本相同的样品，不同之处是其未用测试参数处理，或者如果样品是血浆样品，则其可以来自未受所述感兴趣病症影响的正常志愿者。对照也可以是内部对照。
如本文所用，除非特别指出，则单数形式“一个”、“一种”包括复数形式。因此，例如包含“一个胞外结构域”的化合物包括具有一或多个胞外结构域的化合物。
如本文所用，范围和数量可以用“大约”的特定数值或范围表示。大约也包括精确量。因此，“大约5个碱基”是指“大约5个碱基”也指“5个碱基”。
如本文所用，“任选”是指随后描述的事件或情况发生或不发生，且所述描述包括其中所述事件或情况发生及不发生的情况。例如，任选取代的基团是指该基团不被取代或者被取代。
如本文所用，除非特别指出，则任何保护基团、氨基酸及其它化合物均根据其常用的公认缩写形式或者根据IUPAC-IUB Commission onBiochemical Nomenclature(见(1972)Biochem.11：1726)所述进行缩写。
B.筛选蛋白酶的方法
本发明提供了筛选性质改变、特别是底物特异性和选择性改变的蛋白酶的方法。所述方法还提供了这种改变的蛋白酶，其对于治疗用途呈现出基本未改变的活性或者具有足够活性。本发明提供的方法可以使用蛋白酶修饰及设计修饰的蛋白酶的任何方法。这种方法包括产生文库、使用现有文库的随机方法，也包括定向进化方法。
已经应用了许多选择方案以鉴别具有改变的底物特异性/选择性的蛋白酶，但是每种方法均有限制。本发明提供的方法克服了这种限制。通常，选择方案包括：1)根据蛋白酶结合进行选择或者2)根据蛋白酶催化作用进行选择。利用蛋白酶结合的策略的例子包括例如使用过渡态类似物(TSA)及应用小分子自杀底物的那些策略。TSA是稳定的化合物，其模拟蛋白酶：底物反应的过渡态的电子和结构特征。蛋白酶与底物之间最强的相互作用典型发生在反应的过渡态。TSA用作模型底物以选择具有高度结合亲和性的蛋白酶。TSA不是真正过渡态的理想模拟物且其合成困难(Bertschinger etal.(2005)in Phage display in Biotech.and Drug Discovery(Sidhu S，ed)，pp.461-491)。这种策略鉴别了具有改变的底物特异性的蛋白酶变体，但是这种蛋白酶通常呈现出活性降低，因为蛋白酶催化作用的需求不是该选择方案的一部分。
在另一种策略中，小分子自杀底物(也称作基于机制的抑制剂)用于基于结合而选择蛋白酶。这种自杀底物典型是小分子抑制剂，其共价结合酶的活性位点。这些自杀底物含有反应性亲电体，其与酶亲核体反应形成共价键。这个反应不需要蛋白酶对天然肽键的裂解。典型地，这种抑制剂仅在结合正确的酶及经历正常催化步骤时才产生反应性亲核体(见例如Bertschinger et al.(2005)in Phage display in Biotech.and Drug Discovery(Sidhu S，ed)，pp.461-491)。在许多情况中，底物抑制剂模拟催化作用中涉及的第一过渡态的构象，但是不允许催化循环的完成。结果，这种抑制剂的使用是针对强结合而不是催化作用进行有效选择，导致选择了底物解离减弱的失活酶(Droge et al.(2006)ChemBioChem，7：149-157)。同样，由于其大小和缺乏底物裂解的需求，因此其未重现蛋白酶与天然蛋白质底物的相互作用。
已经尝试了针对催化作用而不是结合进行选择的蛋白酶选择策略(见例如Heinis et al.(2001)，Protein Engineering，14：1043-1052)。针对催化作用进行测定的一种主要限制是反应产物在反应完成后迅速扩散，使得难以分离催化活性酶。因此，针对催化作用进行选择的策略依赖于将底物和酶锚定于噬菌体，由此使其紧密相邻。例如，钙调蛋白已经用作固定剂(Demartis(1999)J Mol.Biol.，286：617-633)。使用钙调蛋白结合肽衍生物将反应底物非共价地锚定于钙调蛋白标记的噬菌体酶上。在催化反应之后，使用抗产物亲和性试剂从非催化活性噬菌体中分离展示反应产物的噬菌体。然而由于底物附着于噬菌体颗粒，由此可以阻碍催化反应。因此，选择蛋白酶的这些及其它方法有一些限制，未鉴别出具有改变的特异性及基本未改变的具有足够治疗应用活性的蛋白酶。本发明的方法解决了这些限制。
本发明提供了选择蛋白酶的方法，以鉴别具有改变的、优化的或者改良的底物特异性的蛋白酶和/或蛋白酶变体。这种蛋白酶被鉴别用于优化及用作治疗性蛋白酶，其可裂解并失活(或者激活)希望的蛋白质靶位，例如疾病或失调的病因学中涉及的蛋白质靶位。在本发明提供的筛选蛋白酶的方法中，候选蛋白酶作为蛋白酶酶反应的稳定中间体复合物而被捕获，然后得以鉴别。所述稳定的中间体复合物典型是共价复合物或者其它复合物，可以从未复合的分子中分离。这种中间体包括例如酰基酶中间体，允许捕获及最终鉴别具有选择的或预定的底物特异性的蛋白酶。通过将蛋白酶集合与由所述蛋白酶裂解的蛋白酶捕获多肽接触，并在裂解时形成稳定复合物而实现蛋白酶的捕获。这种蛋白酶捕获多肽的例子是丝氨酸蛋白酶抑制蛋白、α2巨球蛋白及其它这种分子。所述蛋白酶捕获多肽可以是天然发生的和/或可以是修饰的以针对特定靶底物进行选择。
在实施本发明方法时，将蛋白酶、典型为修饰或突变体蛋白酶集合和/或天然蛋白酶集合与蛋白酶捕获多肽接触，在底物裂解之后所述蛋白酶捕获多肽与蛋白酶反应形成含有捕获的中间体的复合物。这些方法可用于鉴别具有希望的底物特异性/选择性的蛋白酶。为了鉴别具有希望的底物特异性/选择性的蛋白酶，可以在蛋白酶捕获多肽中修饰易断裂键的氨基酸序列和/或反应位点周围序列如反应环序列或者类似序列以模拟希望的靶底物的底物裂解序列。
所述筛选反应通过将蛋白酶集合与蛋白酶捕获多肽在一定条件下接触由此形成稳定的复合物、典型为共价复合物而进行。所述复合物具有足够稳定性，允许其从稳定性较差的复合物和未反应的蛋白酶捕获多肽中分离。
所述蛋白酶捕获多肽可以是可鉴别地标记的或者亲和标记的，以便于鉴别复合物。例如，通过荧光部分、亲和标记或其它这种标记物质对所述蛋白酶捕获多肽进行标记，便于分离蛋白酶-抑制剂复合物及鉴别选择的蛋白酶。可以对选择的蛋白酶进行活性分析以评定蛋白酶解效率和底物特异性。鉴别的或选择的蛋白酶也可以例如通过测序或者其它鉴别方法包括质谱分析法鉴别，或者通过其它标记方法，以鉴别复合物中的选择的蛋白酶。
本发明提供的方法还包括任选地重复筛选步骤，由此对于希望的或预定的特异性/选择性和/或裂解活性的蛋白酶可以进行一次或者可以进行多轮所述方法。例如，蛋白酶选择可包括随机或者根据经验或者系统地修饰被选择的蛋白酶(在靶定区域和/或沿着长度修饰)及重复(一轮、两轮、三轮、四轮或更多轮)将所述蛋白酶集合与一或多种蛋白酶捕获多肽接触的方法。
本发明提供的方法可以是多重的，例如包括两或多个差异标记的或者可差异鉴别的蛋白酶捕获多肽。
在本发明提供的方法中，所述蛋白酶捕获多肽非必需在复合反应中呈现100％或者极高的效率，只要至少可检测的百分比、典型为至少1％、2％、5％、10％、20％、30％、40％、50％、60％、70％、80％、90％或更高百分比可形成稳定的复合物，所述复合物可以从稳定性较差的复合物或者未反应的蛋白酶捕获多肽中分离或者另外鉴别。因此，在反应中发生分配的情况中可选择蛋白酶，其中存在蛋白酶捕获多肽的低于100％抑制作用，例如1％、5％、10％、20％、30％、40％、50％、60％、70％、80％、90％、95％、99％或者更高的蛋白酶催化反应的抑制。在本发明提供的方法中，可以调节选择及其它参数的严格性，例如通过控制反应时间、温度、pH、离子强度和/或文库和底物浓度进行。特异性限制也可以在选择期间加以调节，通过包括竞争剂如含有不希望的底物裂解序列的特定竞争剂或者较广泛类别的竞争剂如人血浆而进行。
本发明提供的方法也可以通过将蛋白酶集合与一种蛋白酶捕获多肽或者不同蛋白酶捕获多肽的混合物接触(例如通过多重)而进行。在使用多个不同蛋白酶捕获多肽的情况中，每个蛋白酶捕获多肽均可以单独及独特地进行标记，由此其可以被可鉴别地检测。这种方法使得可以在一个反应中从蛋白酶集合中分离和鉴别多个蛋白酶。
本发明提供的方法允许立即对多个蛋白酶集合进行筛选以鉴别具有希望的或预定的底物特异性的那些蛋白酶。蛋白酶集合包括包含各种野生型蛋白酶、修饰的蛋白酶、其混合物及其蛋白酶解活性部分的集合的任何蛋白酶集合。可以应用任何集合。所述集合也可以是一组突变体蛋白酶或者含有所述突变的其蛋白酶解活性部分。这种集合包括组合集合，其中所述集合的成员含有多样的突变。所述突变可以是沿着蛋白酶(或其催化活性部分)长度的随机突变，或者靶向特定位置或区域的突变，例如靶向蛋白酶的特异性结合口袋的突变。本发明提供的方法可以鉴别和揭示先前未意识到作为特异性决定簇的非接触的残基(即埋没的残基)。因此，本发明提供的蛋白酶选择技术方法可用于产生具有完全新的特异性和活性的蛋白酶和/或用于优化现有蛋白酶的特异性或活性。
C.蛋白酶捕获多肽
本发明提供的方法中使用的蛋白酶捕获多肽是一种多肽或者含有反应位点的多肽部分，其作为蛋白酶的底物在裂解时导致稳定的蛋白酶-底物中间体复合物形成。通常，这种蛋白酶复合物需要通过所述蛋白酶裂解易断裂键(P1-P1’)以产生捕获的底物-蛋白酶复合物。所述稳定的复合物典型是在蛋白酶与蛋白酶捕获多肽之间通过紧密的相互作用而形成的不可逆复合物，如通过共价键、离子、疏水性或者其它紧密连接形成。由于这种复合物通常可以稳定几小时、几天、几周或者更长时间，由此可以分离该复合物。在一个实例中，稳定的中间体复合物可以是酰基酶中间体，其是在丝氨酸或半胱氨酸蛋白酶与蛋白酶捕获多肽反应时形成的。最通常地，在蛋白酶捕获多肽裂解后，复合物中的快速构象变化使得蛋白酶变形且阻止酰基-酶复合物脱酰作用。因此，用蛋白酶捕获多肽淘选蛋白酶容许针对催化作用的限速步骤(即P1-P1’键的裂解和酶的酰化)进行选择，同时形成易于从集合混合物中分离出非常紧密的(即共价)复合物。
典型地，这种蛋白酶捕获多肽是较大的(100个氨基酸以上)单结构域蛋白，含有由蛋白酶识别的反应位点序列。通常，所述反应位点裂解序列是柔性的、暴露的及较长的大反应环的一部分，使其成为靶底物(Otlewski et al.(2005)The EMBO J.24：1303-1310)，然而，只要蛋白酶捕获剂含有可以由蛋白酶裂解的反应位点序列，从而模拟底物裂解，则其可以用于本发明的方法中。因此，任何大的多肽或者合成产生的多肽均可以用于本发明的方法中，所述多肽含有由蛋白酶裂解的易断裂键，导致在长期稳定复合物中捕获蛋白质。这种蛋白酶捕获多肽的例子是丝氨酸蛋白酶抑制蛋白，如本文所述任何多肽。其它蛋白酶捕获多肽也可以用于在本发明提供的方法中，如其作用机制与丝氨酸蛋白酶抑制蛋白分子相似的那些多肽。这些多肽包括例如合成或者重组产生的丝氨酸蛋白酶抑制蛋白样分子，或者含有丝氨酸蛋白酶抑制蛋白分子的连续片段或者序列的多肽，包括丝氨酸蛋白酶抑制蛋白分子的反应位点环的足够部分。另外，可以使用其抑制机制与丝氨酸蛋白酶抑制蛋白相似的其它蛋白酶抑制剂，例如抑制胱天蛋白酶的杆状病毒p35蛋白(Xu et al.(2001)Nature，410：494-497；Otlewski et al.(2005)The EMBO J.24：1303-1310)。其它蛋白酶捕获多肽包括捕获稳定复合物中蛋白酶的任何多肽，其可易于分离，例如但非限于α2巨球蛋白。
1.丝氨酸蛋白酶抑制蛋白：结构、功能和表达
丝氨酸蛋白酶抑制蛋白(丝氨酸蛋白酶抑制剂)与其它丝氨酸蛋白酶抑制剂(通常为大约少于60个氨基酸)相比是较大的蛋白质分子(大约330-500个氨基酸)的蛋白酶抑制剂。丝氨酸蛋白酶抑制蛋白超家族是最大及最广泛分布的蛋白酶抑制剂。目前在不同动物、痘病毒、植物、细菌和古细菌中已经鉴别了超过1500个丝氨酸蛋白酶抑制蛋白家族成员(Law et al.(2006)Genome Biology，7：216)，迄今为止已经研究了超过30个不同的人丝氨酸蛋白酶抑制蛋白。大多数人丝氨酸蛋白酶抑制蛋白在血液中发现，在此其发挥广泛的调节作用，包括例如炎症、补体、血液凝固和纤维蛋白溶解级联反应。丝氨酸蛋白酶抑制蛋白也在细胞内发挥细胞保护作用，例如调节细胞毒性蛋白酶的不适当释放。尽管大多数丝氨酸蛋白酶抑制蛋白对于蛋白酶活性具有抑制作用，但是一些丝氨酸蛋白酶抑制蛋白具有其它非抑制性作用，例如但非限于激素转运、皮质类固醇激素结合球蛋白、及血压调节作用(Silverman et al.(2001)JBC，276：33293-33296)。非抑制性丝氨酸蛋白酶抑制蛋白是类固醇结合球蛋白和卵清蛋白。典型地，丝氨酸蛋白酶抑制蛋白抑制丝氨酸蛋白酶的作用，但是已经鉴别出几个丝氨酸蛋白酶抑制蛋白是木瓜蛋白酶样半胱氨酸蛋白酶或胱天蛋白酶的抑制剂(Whisstock et al.(2005)FEBS Journal，272：4868-4873)。
丝氨酸蛋白酶抑制蛋白家族成员的序列相同性较差，然而，其结构是高度保守的。例如，丝氨酸蛋白酶抑制蛋白家族成员与丝氨酸蛋白酶抑制蛋白α1-抗胰蛋白酶呈现大约30％氨基酸序列同源性，且具有保守的三级结构。在结构上，丝氨酸蛋白酶抑制蛋白由三个β折叠(A、B和C)和8-9个α-螺旋(A-I)组成，其组成上部β-桶结构域和下层螺旋结构域。这两个结构域由5链B-折叠A桥连，其是丝氨酸蛋白酶抑制蛋白的主要结构特征(Huntington et al.(2003)，J.Thrombosis and Haemostasis，1：1535-1549)。丝氨酸蛋白酶抑制蛋白是亚稳定蛋白，由此其在活性形式仅是部分稳定的；其需要蛋白酶以采取完全稳定构象。称作反应位点环(RSL)的环可以改变丝氨酸蛋白酶抑制蛋白分子的构象。RSL是一段大约17个氨基酸残基的暴露序列，其在A和Cβ-折叠之间的区域中从分子的顶部突出。RSL作为蛋白酶识别位点，通常含有唯一的蛋白酶特异性决定簇。丝氨酸蛋白酶抑制蛋白结构的最稳定形式是RSL-裂解的形式。在蛋白酶裂解之后，RSL的氨基末端部分插入β-折叠A的中心，成为六链β-折叠的第四链。这个构象改变称作从“紧张(stressed)”至“松弛(relaxed)”(或者S至R)转换。这种构象特征在于分子的热稳定性增加，这是由于五链β-折叠A向六链反平行形式的重构所致(Lawrence et al.(2000)，J Biol.Chem.，275：5839-5844)。换句话说，丝氨酸蛋白酶抑制蛋白的天然结构与潜在的中间体等价，所述潜在的中间体仅在经蛋白酶裂解之后转变为更稳定的结构(Law et al.(2006)GenomeBiology，7：216)。
典型地，丝氨酸蛋白酶抑制蛋白靶向丝氨酸蛋白酶，但是一些丝氨酸蛋白酶抑制蛋白使用相似机制抑制半胱氨酸蛋白酶。RSL环决定哪种蛋白酶被靶向抑制，因为其提供了所述靶蛋白酶的假底物。实质上，特定丝氨酸蛋白酶抑制蛋白的抑制特异性由RSL序列介导，其是丝氨酸蛋白酶抑制蛋白中的最可变区域(Travis et al.(1990)Biol.Chem.Hoppe Seyler，371：3-11)。RSL模拟蛋白酶的底物识别序列，因此含有以...Pn-P3-P2-P1-P1’-P2’-P3’-P’n...编号的反应位点，其中所述反应位点是P1与P1’之间的易断裂键。对于成熟的α1-抗胰蛋白酶，在Met358-Ser359键(相应于SEQ ID NO：1所示氨基酸序列的Met382和Ser389氨基酸)发生P1-P1’键的裂解。所述蛋白酶上残基的相应结合位点是...Sn-S3-S2-S1-S1’，S2’，S3’，Sn’-...。在本发明提供的方法中，对RSL序列进行修饰以从展示文库中选择底物特异性改变的蛋白酶，如下文详细论述。
2.蛋白酶催化、丝氨酸蛋白酶抑制蛋白的抑制机制及酰基酶中间体的形成
本发明提供的蛋白酶选择方法利用多肽捕获蛋白酶的能力鉴别具有改变的底物特异性的蛋白酶，如通过丝氨酸蛋白酶抑制蛋白所例证。蛋白酶催化机制在蛋白酶解酶的不同类别之间略为不同，所述蛋白酶例如是丝氨酸蛋白酶、半胱氨酸蛋白酶、天冬氨酸蛋白酶、苏氨酸蛋白酶或者金属蛋白酶。例如，丝氨酸肽酶具有包含在活性中心内的丝氨酸残基，天冬氨酸肽酶催化中心内具有两个天冬氨酸，半胱氨酸型肽酶具有半胱氨酸残基，苏氨酸型肽酶具有苏氨酸残基，金属肽酶在催化机制中使用金属离子。通常，形成共价中间体的那些蛋白酶家族是本发明提供的蛋白酶选择方法的靶。这些包括例如丝氨酸和半胱氨酸蛋白酶家族的成员。例如，对于丝氨酸蛋白酶而言，催化作用的第一步是在底物与蛋白酶的催化中心内的丝氨酸之间形成酰基酶中间体。这种共价中间体的形成通过负电荷四面体过渡态中间体行进，然后底物的P1-P1’肽键被裂解。在第二步或脱酰作用期间，酰基-酶中间体由水分子水解，释放肽及恢复所述酶的Ser-羟基。也包含四面体过渡态中间体形成的脱酰作用通过酰化的反向反应途径行进。对于脱酰作用，水分子代替Ser残基成为攻击亲核体。丝氨酸蛋白酶催化中心的His残基提供了通用碱，并接受反应性Ser的OH基团。
丝氨酸蛋白酶抑制蛋白使用上述S至R转换抑制丝氨酸和半胱氨酸靶蛋白酶的催化反应。其作用机制在蛋白酶抑制剂中是独特的，通过在脱酰化之前破坏蛋白酶的活性位点，从而不可逆地阻碍蛋白酶解，随后形成酰基-酶中间体(Otlewski et al.(2005)The EMBO Journal，24：1303)。丝氨酸蛋白酶抑制蛋白与蛋白酶反应的动力学模型与底物的蛋白酶解的相同(见例如图1；Zhou et al.(2001)J.Biol.Chem.，276：27541-27547)。在与靶蛋白酶相互作用之后，丝氨酸蛋白酶抑制蛋白最初通过P1-P1’易断裂键两侧RSL中的的残基的相互作用而形成非共价Michaelis样复合物(Silverman et al.(2001)，J.Biol.Chem.，276：33293-33296)。蛋白酶活性位点中的丝氨酸残基(对于丝氨酸蛋白酶而言)攻击P1-P1’键，促进肽键裂解及丝氨酸残基与P1残基的主链羰基之间共价酯键的形成。在RSL被裂解之后，RSL插入丝氨酸蛋白酶抑制蛋白分子的β-折叠A中。插入的第一个残基是P14(即成熟α1-抗胰蛋白酶中第345位氨基酸，其相应于SEQ ID NO：1所示氨基酸序列中的T369氨基酸)，随后是RSL的柔性铰链区(P15-P9)(Buck et al.(2005)Mol.Biol.Evol.，22：1627-1634)。RSL的插入转运与其共价结合的蛋白酶，导致蛋白酶的构象改变，特征在于变形的活性位点(见图1)以及丝氨酸蛋白酶抑制蛋白转换为“松弛”状态。蛋白酶的构象变化改变了活性位点的催化三联体，由此P1侧链从S1口袋中除去。构象重排的最终结果是捕获酰基酶中间体(Silverman et al.(2001)，J.Biol.Chem.，276：33293-33296)。
酰基酶的形成对于丝氨酸蛋白酶抑制蛋白相互作用是重要的，因此丝氨酸蛋白酶抑制蛋白典型特异于在催化作用中具有酰基酶中间体的蛋白酶类别。这些类别的蛋白酶是丝氨酸蛋白酶家族的主要成员，包括胰凝乳蛋白酶超家族中的那些蛋白酶及蛋白酶的枯草杆菌蛋白酶超家族中的那些蛋白酶，这些蛋白酶在下文详细描述。此外，丝氨酸蛋白酶抑制蛋白对于半胱氨酸蛋白酶也具有反应性，所述半胱氨酸蛋白酶包括例如丝氨酸蛋白酶的木瓜蛋白酶家族和胱天蛋白酶家族中的那些蛋白酶。典型地，丝氨酸蛋白酶抑制蛋白不抑制金属-、苏氨酸或天冬氨酸家族的蛋白酶。例如，丝氨酸蛋白酶抑制蛋白与金属蛋白酶的相互作用不产生共价捕获的中间体，而是金属蛋白酶使所述抑制剂裂解而不形成任何复合物(Li et al.(2004)Cancer Res.64：8657-8665)。
因此，尽管大多数丝氨酸蛋白酶抑制蛋白抑制胰凝乳蛋白酶家族的丝氨酸蛋白酶，但是确实存在抑制半胱氨酸蛋白酶的跨类别(cross-class)抑制剂。所述跨类别抑制剂是抑制胱天蛋白酶1的病毒丝氨酸蛋白酶抑制蛋白CrmA和PI9(SEPRINB9)及抑制木瓜蛋白酶样半胱氨酸蛋白酶(包括组织蛋白酶L、K和S)的SCCA1(SEPRINB3)。丝氨酸蛋白酶抑制蛋白介导的丝氨酸蛋白酶抑制机制看起来适于半胱氨酸蛋白酶。然而，差异之处是动力学捕获的中间体是硫羟酸酯而不是在丝氨酸蛋白酶情况中的氧酯(oxyester)(Silverman et al.(2001)J.Biol.Chem.，276：33293-33296)。稳定的共价硫羟酸酯型键的存在通过检测到SCCA1与组织蛋白酶S之间SDS稳定复合物而支持(Silverman et al.(2001)J.Biol.Chem.，276：33293-33296；Schick etal.(1998)Biochemistry，37：5258-5266)。
根据丝氨酸蛋白酶抑制蛋白-蛋白酶对，丝氨酸蛋白酶抑制蛋白-蛋白酶对高度稳定持续几周至几年，然而最后终会发生解离以产生正常蛋白酶解产物(即裂解的丝氨酸蛋白酶抑制蛋白和活性蛋白酶；见例如Zhou et al.(2001)J.Biol.Chem.，276：27541-27547)。另外，如果RSL环未足够快地插入蛋白酶中，则反应直接行进至裂解产物。这个现象称作分配(partitioning)且反映出可以发生的分支途径的存在，导致稳定抑制复合物或者丝氨酸蛋白酶抑制蛋白至底物的转换，如图1所示，抑制复合物的形成相对非抑制途径(Lawrence et al.(2000)，J.Biol.Chem.，275：5839-5844)。丝氨酸蛋白酶抑制蛋白的分配可以通过改变RSL环、特别是起始环插入的RSL的铰链区中的残基(即P14)而调节，或者通过改变丝氨酸蛋白酶抑制蛋白与其最佳相互作用的蛋白酶而调节。例如，丝氨酸蛋白酶抑制蛋白纤溶酶原激活物抑制剂-1(PAI-1)的抑制活性在蛋白酶uPA、tPA与凝血酶之间不同，靶向偏爱uPA和tPA。另外，RSL环在例如铰链区的P14位置的变化改变了PAI-1的靶向偏爱性：突变为带电荷的氨基酸(即Arg、Lys、Asp、Glu)降低了PAI-1对于uPA、tPA和凝血酶的抑制活性；突变为中性氨基酸(即His、Tyr、Gln、Asn)或者突变为无侧链的Gly导致PAI-1对于tPA和凝血酶的抑制活性与对于uPA相比降低10-100倍；突变为疏水性氨基酸与野生型PAI-1相比不改变PAI-1的抑制活性(Lawrence et al.(2000)，J.Biol.Chem.，275：5839-5844)。
在丝氨酸蛋白酶抑制蛋白介导的对蛋白酶催化的成功抑制中重要的因子是RSL环的长度，其必须是精确长度以保证丝氨酸蛋白酶抑制蛋白与蛋白酶的相互作用提供了丝氨酸蛋白酶抑制蛋白本体(body)与蛋白酶之间的杠杆作用，允许催化性丝氨酸从活性位点的位移及蛋白酶的变形(Zhou etal.(2001)J.Biol.Chem.，276：27541-27547；Huntington et al.(2000)Nature，407：923-926)。实际上，丝氨酸蛋白酶抑制蛋白本体使得所述蛋白酶被变形。大多数丝氨酸蛋白酶抑制蛋白具有RSL，所述RSL长度为17个残基，而仅几个丝氨酸蛋白酶抑制蛋白经鉴别具有16个残基的环(即α2-抗纤溶酶、C1-抑制剂和CrmA)。具有18个残基环的α2-抗纤溶酶变体丝氨酸蛋白酶抑制蛋白已经从出血性疾病患者中鉴别，但是这种变体不是有功能的抑制性丝氨酸蛋白酶抑制蛋白(Zhou et al.(2001)J.Biol.Chem.，276：27541-27547)。因此，丝氨酸蛋白酶抑制蛋白抑制机制可以适应缩短的RSL，但是不适应加长的RSL(Zhou et al.(2001)J.Biol.Chem.，276：27541-27547)。除了丝氨酸蛋白酶抑制蛋白家族成员的环长度的保守性之外，丝氨酸蛋白酶抑制蛋白的RSL通常也保留保守的铰链区(P15-P9)组成，且典型不含有带电荷的或大的P残基。
a.丝氨酸蛋白酶抑制蛋白的例子
本发明提供的方法中使用的丝氨酸蛋白酶抑制蛋白可以是任何丝氨酸蛋白酶抑制蛋白多肽，包括但非限于重组产生的多肽、合成产生的多肽和从细胞、组织和血液中提取的丝氨酸蛋白酶抑制蛋白。丝氨酸蛋白酶抑制蛋白还包括等位基因变体及来自不同物种的多肽，包括但非限于来自人及非人动物、痘病毒、植物、细菌和古细菌。典型地，丝氨酸蛋白酶抑制蛋白的等位基因或物种变体与天然或野生型丝氨酸蛋白酶抑制蛋白具有大约或者至少40％、50％、60％、70％、80％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％的不同。人丝氨酸蛋白酶抑制蛋白包括本发明提供的任何丝氨酸蛋白酶抑制蛋白(例如下表2所示)、等位基因变体同工型、从核酸合成的分子、从人组织、细胞或血液分离的蛋白质，及任何人丝氨酸蛋白酶抑制蛋白多肽的修饰形式。丝氨酸蛋白酶抑制蛋白还包括截短的多肽片段，只要存在足够的RSL环部分以介导与蛋白酶的相互作用及形成共价酰基酶中间体。
典型地，本发明提供的方法中使用的丝氨酸蛋白酶抑制蛋白是抑制性丝氨酸蛋白酶抑制蛋白或其片段，能在丝氨酸蛋白酶抑制蛋白与蛋白酶之间形成共价酰基酶中间体。通常，这种丝氨酸蛋白酶抑制蛋白用于选择由丝氨酸蛋白酶抑制蛋白正常靶向的蛋白酶，在此发生接近于蛋白酶的完全抑制并且在抑制性复合物与裂解的丝氨酸蛋白酶抑制蛋白底物之间的分配被最小化。表3示出丝氨酸蛋白酶及其关联丝氨酸蛋白酶抑制蛋白抑制剂的实例。预期这种丝氨酸蛋白酶抑制蛋白/蛋白酶对具有高缔合常数或者抑制作用二级速率常数并且低或不分配为非抑制性复合物。例如，t-PA的主要生理抑制剂是丝氨酸蛋白酶抑制蛋白PAI-1，一种大约50kD的糖蛋白(Pannekoek et al.(1986)EMBO J.，5：2539-2544；Ginsberg et al.，(1980)J.Clin.Invest.，78：1673-1680；and Carrell et al.In.：Proteinase Inhibitors，Ed.Barrett，A.J.et al.，Elsevier，Amsterdam，pages 403-420(1986)。然而，其它丝氨酸蛋白酶抑制蛋白/蛋白酶对也可以用于本发明提供的方法中，即使缔合常数较低而分配较高。例如，其他丝氨酸蛋白酶抑制蛋白如C1酯酶抑制剂和α-2-抗纤溶酶与tPA的缔合常数较PAI-1的低几个数量级(Ranby et al.(1982)Throm.Res.，27：175-183；Hekman et al.(1988)Arch.Biochem.Biophys.，262：199-210)，但是这些丝氨酸蛋白酶抑制蛋白仍抑制tPA(见例如Lucore etal.(1988)Circ.77：660-669)。
表3
| 丝氨酸蛋白酶 | 关联丝氨酸蛋白酶抑制蛋白 |
| 激活的C蛋白 | C蛋白抑制剂PAI-1 |
| C1酯酶 | C1酯酶抑制剂 |
| 组织蛋白酶G | α-1-抗胰蛋白酶α-1-抗胰凝乳蛋白酶 |
| 类胰凝乳蛋白酶 | α-1-抗胰凝乳蛋白酶 |
| 胰凝乳蛋白酶 | α-1-抗胰凝乳蛋白酶α-2-抗纤溶酶制酶蛋白 |
| 凝血因子(VIIa，Xa，XIa，XIIa) | 抗凝血酶IIIC1酯酶抑制剂 |
| 弹性蛋白酶 | α-1-抗胰蛋白酶 |
| 激肽释放酶 | C1酯酶抑制剂α-1-抗胰蛋白酶 |
| 纤溶酶 | α-2-抗纤溶酶 |
| 凝血酶 | 抗凝血酶III肝素辅因子II |
| tPA | PAI-1，PAI-2，PAI-3 |
| 胰蛋白酶 | α-1-抗胰蛋白酶生长激素调节蛋白 |
| 胰蛋白酶样蛋白酶 | 蛋白酶微管连接蛋白I |
| u-PA | PAI-1，PAI-2，PAI-3 |
因此，在本发明提供的方法中用于选择蛋白酶的丝氨酸蛋白酶抑制蛋白通常产生这样的反应产物，其中80％、90％、93％、94％、95％、96％、97％、98％、99％或100％所述反应产物是抑制性复合物的形成。在一些情况中，在本发明提供的方法中可以发生丝氨酸蛋白酶抑制蛋白与蛋白酶之间的分配增加，例如如果本发明方法中使用的丝氨酸蛋白酶抑制蛋白不是最佳靶向蛋白酶。因此，在本发明提供的方法中，丝氨酸蛋白酶抑制蛋白可用于选择蛋白酶，其中所得反应导致为或大约为20％、30％、40％、50％、60％、70％、75％或更多的稳定抑制性复合物，其余的产物是裂解的丝氨酸蛋白酶抑制蛋白底物。当发生分配时可被改变以优化蛋白酶选择的因素包括例如增加的丝氨酸蛋白酶抑制蛋白浓度和增加的反应时间。在一些情况中，如下文讨论的其它非抑制性丝氨酸蛋白酶抑制蛋白或其突变体可以用于本发明提供的方法中，只要进行选择的靶蛋白酶能与丝氨酸蛋白酶抑制蛋白底物相互作用产生可以被捕获的共价抑制性复合物。
i.PAI-1
蛋白酶选择方法中使用的丝氨酸蛋白酶抑制蛋白例如是纤溶酶原激活物抑制剂-1(PAI-1)或其变体。PAI-1是组织纤溶酶原激活物(t-PA)和尿激酶或者尿纤溶酶原激活物(u-PA)的主要抑制剂，其是参与由纤溶酶原激活所致的纤维蛋白溶解中的蛋白酶。PAI-1对于t-PA和u-PA具有大约2×107M-1s-1的二级速率常数。PAI-1参与肿瘤浸润、纤维蛋白溶解、细胞迁移、组织重塑、组织内卷、排卵、炎症、滋养层浸润及恶性转化(Salonen et al.(1988)JBiol.Chem.，264：6339-6343)。PAI-1主要由内皮产生，但是也由其它组织类型例如脂肪组织分泌。其它相关的纤溶酶原激活物抑制剂包括PAI-2和PAI-3。例如，PAI-2也是一种u-PA和t-PA抑制剂，但是由胎盘分泌，且典型仅在妊娠期间大量存在。
PAI-1是具有SEQ ID NO：11所示前体序列的单链糖蛋白，包括23个氨基酸的信号序列，当裂解时产生379个氨基酸的成熟序列。与其它丝氨酸蛋白酶抑制蛋白一样，PAI-1在由蛋白酶在其位于Arg346-Met347(即相应于SEQ ID NO111所示前体序列的Arg369和Met370氨基酸)的P1-P1’反应位点裂解之后由潜伏形式转换为活性形式，从而导致稳定的共价复合物形成及结合的蛋白酶失活。然而，与其它丝氨酸蛋白酶抑制蛋白不同，PAI-1采用潜伏过渡态自发产生失活的、高度稳定的但共价完整的形式，从而RSL的残基P15至P4插入β-折叠A中形成β-折叠的第四链(即s4A)，残基P3至P10’在分子表面形成突出的环(De Taeye et al.(2003)J Biol.Chem.，278：23899-23905)。因此，活性PAI-1在37℃相对不稳定，呈现仅2.5小时的半衰期，之后自发转变为潜伏构象。然而，这种潜伏形式可以通过变性而再激活，例如通过十二烷基硫酸钠、胍氯化物(guanidinium chloride)和脲变性(Declerek et al.(1992)J.Biol.Chem.，267：11693-11696)及加热(Katagiri etal.(1988)Eur J.Biochem.，176：81-87)变性。PAI-1的活性形式也通过与玻连蛋白相互作用而稳定化。已经鉴别了不能经历转变为潜伏构象及因此在温度和pH升高时更长期更稳定的突变PAI-1(见例如Berkenpas et al.(1995)The EMBO J.，14：2969-2977)。
已经对丝氨酸蛋白酶(即t-PA或u-PA)和/或抑制性丝氨酸蛋白酶抑制蛋白(即PAI-1)进行修饰以调节或改变抑制作用的二级速率常数以便使得蛋白酶对其关联丝氨酸蛋白酶抑制蛋白抑制剂或其变体具有抗性，例如用于需要野生型蛋白酶活性的治疗应用中(见例如U.S.Patent Serial Nos.5,866,413；5,728,564；5,550,042；5,486602；5,304,482)。
ii.抗凝血酶(AT3)
用于本发明方法中的丝氨酸蛋白酶抑制蛋白或其变体的另一例子是抗凝血酶(AT3)。AT3也是丝氨酸蛋白酶抑制蛋白家族的成员，使得许多酶失活，包括例如来自凝血系统的那些酶，例如但非限于因子X、因子IX、因子II(凝血酶)、因子VII、因子XI和因子XII。典型地，抗凝血酶主要在血液中发现，在此其通过阻断凝血酶的功能而防止或抑制血液凝固。AT3的活性通过一或多种辅因子、典型为肝素的存在而增加。在与肝素相互作用时，AT3经历构象重排，包括环从丝氨酸蛋白酶抑制蛋白结构中排出及P1暴露，导致形成具有暴露的蛋白酶可及构象的AT3结构。另外，肝素既可结合蛋白酶也可结合抑制剂，从而加速抑制机制(Law et al.(2006)GenomeBiology，7(216)：1-11)。
AT3的基因序列编码跨越七个外显子的DNA，所述DNA编码SEQ IDNO：5所示前体蛋白。相应于SEQ ID NO：5所示序列1-32位氨基酸的信号序列的裂解导致产生432个氨基酸的、分子量为大约58000道尔顿的成熟蛋白质。其中6个氨基酸是半胱氨酸，其导致三个分子内二硫键形成。AT3的RSL中P4-P2’位含有氨基酸残基IAGRSL(SEQ ID NO：478)，其相应于SEQ ID NO：5所示氨基酸序列中第422-427位氨基酸，其中在反应位点P1-P1’的裂解发生在Arg425-Ser426氨基酸之间。
3.其它蛋白酶捕获多肽
本领域已知或者可以鉴别其它蛋白酶捕获多肽，其呈现与丝氨酸蛋白酶抑制蛋白相似的抑制机制(例如靶底物由蛋白酶裂解产生稳定的中间体及蛋白酶结构的构象变化)。这种蛋白酶捕获多肽预期用于本发明提供的方法中。这种蛋白酶捕获多肽的例子是p35。另外，本发明提供的方法中可以使用由蛋白酶裂解导致在长期稳定复合物中捕获蛋白酶的任何其它分子。
a.p35
例如，杆状病毒p35蛋白(SEQ ID NO：473)是广谱胱天蛋白酶抑制剂，其可以这种方式抑制胱天蛋白酶(Xu et al.(2001)Nature 410：494-497；Xu etal.(2003)J.Biol.Chem.278(7)：5455-5461)。胱天蛋白酶对p35的P1-P1’键的裂解(在胱天蛋白酶裂解位点DQMD87)在p35环的氨基节段(Asp87)与胱天蛋白酶催化三联体的半胱氨酸残基(胱天蛋白酶-8中Cys350)之间产生共价硫酯中间体。在硫酯键形成时，所述蛋白酶经历构象改变，允许裂解的环的氨基节段埋入胱天蛋白酶中，而在第2位含有Cys残基的p35的N末端插入胱天蛋白酶活性位点中，因此阻断胱天蛋白酶-8中His 317残基的溶剂可及性。对水解性水分子的不可及性因此防止随后的硫酯键水解。
除了p35之外，相似的病毒胱天蛋白酶抑制剂包括但非限于p49(SEQID NO：491)和丝氨酸蛋白酶抑制蛋白CrmA牛痘基因(SEQ ID NO：492)。p49抑制剂呈现与p35相似的胱天蛋白酶抑制机制，其中在p49胱天蛋白酶识别序列TVTD94裂解时与胱天蛋白酶的活性位点形成稳定的硫酯键。
使用本发明提供的方法进行筛选的靶底物可以包括病毒胱天蛋白酶抑制剂多肽，如p35、p49或CrmA多肽。修饰本发明提供的丝氨酸蛋白酶抑制蛋白的RSL环的方法可以容易地适应于病毒胱天蛋白酶抑制剂多肽的修饰。例如，可以对p35RSL中裂解的靶位点进行修饰，以选择对于靶底物具有改变的反应性或特异性的蛋白酶。在野生型p35中，在84-87(DQMD87)氨基酸位置发现胱天蛋白酶识别。对于病毒胱天蛋白酶抑制剂多肽进行的修饰因此可包括改变裂解序列和/或周围氨基酸残基的修饰。例如，这种修饰的胱天蛋白酶抑制剂多肽例如p35、p49或CrmA多肽可以设计为模拟希望的靶底物的裂解序列，例如疾病或病症的病因学中涉及的靶底物。在本发明提供的方法中可以对病毒胱天蛋白酶抑制剂多肽的RSL环序列进行任何修饰。
本发明提供的方法中使用的病毒胱天蛋白酶抑制剂多肽如p35、p49或CrmA多肽可以是任何病毒胱天蛋白酶抑制剂多肽，包括但非限于重组产生的多肽、合成产生的多肽及通过杆状病毒纯化方法产生的p35或p49多肽。病毒胱天蛋白酶抑制剂多肽也包括多肽的等位基因变体，如p35、p49或CrmA多肽变体。
b.α巨球蛋白(aM)
蛋白酶的α巨球蛋白(aM)家族包括蛋白酶抑制剂，如举例的蛋白酶抑制剂α-2-巨球蛋白(a2M，SEQ ID NO：490)，预期在本发明提供的方法中用作蛋白酶捕获剂。aM分子抑制所有类别的蛋白酶。aM蛋白酶捕获剂特征在于相似的抑制机制，包括蛋白酶对抑制剂诱饵区域的裂解。所述诱饵区域是对蛋白酶解敏感的节段，在裂解时aM分子中发生构象改变，导致蛋白酶周围结构折拢(collapse)。例如在SEQ ID NO：490所示a2M序列中的诱饵区域相应于第690-728位氨基酸。在获得的aM-蛋白酶稳定复合物中，蛋白酶的活性位点在空间上被隔离，从而降低对正常蛋白酶底物的可及性。典型地，捕获的蛋白酶保留对于小肽底物的活性，但是丧失其与大蛋白质底物或抑制剂相互作用的能力。另外，aM分子特征在于存在反应性硫羟酸酯，通过硫羟酸酯与胺的反应使得抑制能力失活。另外，在诱饵区域裂解时发生的构象变化暴露了保守的COOH-末端受体结合结构域(RBD)。RBD序列的暴露便于从循环中除去aM-蛋白酶复合物。
4.蛋白酶捕获竞争剂
在本发明提供的方法中可以使用竞争剂，以调节被选择的蛋白酶对于靶底物的特异性和选择性限制。可以将竞争剂在任何时间与蛋白酶或其集合接触，如在蛋白酶与希望的蛋白酶捕获多肽接触之前或之后进行接触，或者竞争剂与希望的蛋白酶捕获多肽可以同时与蛋白酶接触。竞争剂可以是特异性竞争剂或者广谱竞争剂。
特异性竞争剂被设计为模拟预定的非靶底物，从而作为预定的潜在脱靶物(off-target)。典型地，这种竞争剂未被标记，以便不选择形成的稳定蛋白酶复合物。另外，这种竞争剂以大大超过选择方案中使用的设计的蛋白酶捕获多肽的量加入，典型以过量的摩尔量加入，由此所述竞争剂结合所述集合中不希望的蛋白酶。在特异性竞争的一个实例中，将两个不同的蛋白酶捕获多肽(每个多肽均设计为模拟不同的底物识别)与蛋白酶集合接触，其中仅一个蛋白酶捕获多肽是可检测地标记的。例如，竞争剂可包括设计为具有模拟非靶底物的裂解序列的反应位点的多肽蛋白酶捕获剂。因此，竞争剂如丝氨酸蛋白酶抑制蛋白可以设计为其P4-P1’RSL残基由预定的非靶底物的裂解序列置换。所述竞争剂可以与蛋白酶捕获多肽组合用于本发明方法中，例如与另一种丝氨酸蛋白酶抑制蛋白多肽组合使用，其RSL序列已经修饰为在P4-P1’位含有模拟希望的或预定的靶底物的裂解序列的氨基酸，且被标记以进行分离。因此，这两种蛋白酶捕获多肽均选择对靶裂解序列或非靶裂解序列呈现选择性的蛋白酶，但是仅那些呈现希望的靶底物特异性且被可检测地标记的稳定的蛋白酶复合物可以从反应中分离。特异性竞争剂的其它实例包括例如天然蛋白酶捕获多肽，其反应位点已经在本发明提供的方法中被修饰。实施例6例证了这种策略，其中血浆纯化的AT3丝氨酸蛋白酶抑制蛋白用作修饰的丝氨酸蛋白酶抑制蛋白AT3SLGR-KI的竞争剂。
广谱竞争剂也可以用于本发明提供的方法中，以限制被选择的蛋白酶的特异性和选择性。广谱竞争剂的例子包括例如含有多种天然蛋白酶抑制剂的人血浆或人血清。或者，可以产生蛋白酶捕获多肽的广泛小分子文库，其中P2、P3或P4的每个位置均是不同的，例如Acxxx-Thiaphine文库。
5.变体蛋白酶捕获多肽
已经修饰为在其反应位点具有改变的裂解序列的蛋白酶捕获多肽可以用于本发明提供的方法中，以用希望的或预定的靶底物选择蛋白酶。因此，蛋白酶捕获剂在其作为公认的蛋白酶裂解位点的序列区域被修饰，以选择对于靶底物具有改变的反应性或特异性的蛋白酶。例如，丝氨酸蛋白酶抑制蛋白可以被修饰为在RSL环的易断裂键或其周围具有改变的裂解序列。在另一实例中，a2M可以在其诱饵区域中被修饰以具有改变的裂解序列。这种修饰的蛋白酶捕获剂可以设计为模拟希望的靶底物的裂解序列，例如疾病或病症的病因学中涉及的靶底物。
丝氨酸蛋白酶抑制蛋白分子的RSL环序列中的任何修饰均可以在本发明提供的方法中产生。举例的野生型丝氨酸蛋白酶抑制蛋白的RSL序列的对比在下表4中示出。在下表中，P15至P5’位的编号是就成熟α1-抗胰蛋白酶分子而言(相应于SEQ ID NO：1所示氨基酸序列的第367-387位氨基酸)。RSL环序列的特性为本领域技术人员所已知和/或可以通过序列对比而确定，例如通过与下表4示出的丝氨酸蛋白酶抑制蛋白对比。
表4：RSL环序列对比＊
| 丝氨酸蛋白酶抑制蛋白 | RSL环序列 | SEQ IDNO |
| 343P15 P10 P4 P1P1′P5′363 | ||
| Manduca sexta丝氨酸蛋白酶抑制蛋白1BManduca sexta丝氨酸蛋白酶抑制蛋白1Kα1-抗胰凝乳蛋白酶抗凝血酶-IIIPAI-IIα1-抗胰蛋白酶PAI-IPAI-III卵清蛋白 | EGAEAAAANAFGIVPKSLILYEGAEAAAANAFKITTYSFHFVEGTEASAATAVKITLLSALVEEGSEAAASTAVVIAGRSLNPNEGTEAAAGTGGVMTGRTGHGGKGTEAAGAMFLEAIPMSIPPESGTVASSSTAVIVSARMAPEESGTRAAAATGTIFTFRSARLNAGREVVGSAEAGVDAASVSEE | 397398399400401402403404405 |
*根据Ye et al.(2001)Nature Structural Biology 8：979所述改编
因此，可以修饰丝氨酸蛋白酶抑制蛋白的RSL环中相应于丝氨酸蛋白酶抑制蛋白反应位点中的任一或多个氨基酸的氨基酸序列(即相应于如上表4示出的P15至P5’位的任一或多个氨基酸)。典型地，作为RSL环序列的铰链区一部分的氨基酸(即相应于P15-P9位的氨基酸)未被修饰。在一个实例中，P1和/或P1’位中的一或多个氨基酸被修饰以相应于在易断裂键两侧的那些氨基酸。在另一实例中，相应于反应位点位置P4-P2’的任一或多个氨基酸被修饰。例如，PAI-1的P4-P1’是VSARM(SEQ ID NO：378)，其中在R(P1)与M(P1’)氨基酸之间发生裂解。可以修饰VSARM序列的任一或多个氨基酸以修饰PAI-I的裂解序列以选择具有改变的特异性的蛋白酶。实施例1例证了PAI-I的修饰，其中反应位点环中的VSARM序列被修饰为RRARM(SEQ ID NO：379)。在另一实例中，反应位点环VSARM序列可以被修饰为已知的有效肽底物PFGRS(SEQ ID NO：389)。这种突变PAI-1的例子如SEQ ID NO：610和611示出。
在另一实例中，可以在抗凝血酶III(AT3)的RSL中产生修饰。例如，AT3的P4-P1’是IAGRSL(SEQ ID NO：478)，其中裂解发生在R(P1)与S(P1’)氨基酸之间。可以对IAGRSL序列的任一或多个氨基酸进行修饰以修饰AT3的裂解序列以选择具有改变的特异性的蛋白酶。实施例6和7例证了AT3的修饰，其中反应位点环中的IAGRSL序列被修饰为RRVRKE(SEQ ID NO：498)。在另一实例中，反应位点环中的IAGRSL氨基酸序列可以被修饰为SLGRKI(SEQ ID NO：479)。其它修饰的AT3多肽含有IAGRSL氨基酸序列由氨基酸序列SKGRSL(SEQ ID NO：501)或者氨基酸序列PRFKII(SEQ IDNO：503)置换。这种突变AT3分子的例子以SEQ ID NO：497、499、500和502示出。
或者及如果需要，则可以以一次一个、一次两个、一次三个等方式对P4-P2’位置的任一或多个氨基酸进行修饰，获得的修饰的丝氨酸蛋白酶抑制蛋白可以在连续轮的选择中单独检测，以优化在每个修饰的位置呈现底物特异性和/或选择性的蛋白酶。
在大多数情况中，置换野生型丝氨酸蛋白酶抑制蛋白反应位点环或者另一蛋白酶捕获剂中类似序列中的氨基酸残基的氨基酸残基是基于在希望的靶底物中的裂解序列而选择。靶底物蛋白是正常参与病理学中的底物，其中在指定底物序列裂解靶蛋白作为对病理进行治疗的方式(见例如美国专利公开号US 2004/0146938，US2006/0024289，US2006/0002916，及临时申请系列号60/729,817)。例如，靶蛋白可以是类风湿性关节炎(即TNFR)、败血症(即C蛋白)，致肿瘤原性(即生长因子受体，如VEGFR)或者炎症(即补体蛋白)中涉及的蛋白质。靶底物也可以是病毒蛋白，由此在病毒蛋白裂解时，所述病毒不能感染细胞。下表5举例示出了靶底物。
靶蛋白内的裂解位点是已知的或者可易于鉴别。通过如下标准鉴别靶蛋白内的裂解位点：1)其位于蛋白质的暴露表面；2)其位于缺乏二级结构的区域中(即不在螺旋的P折叠中)，如通过结构预测算法的原子结构确定(这些区域倾向于是蛋白质表面上的环或者细胞表面受体上的柄)；3)其位于基于其已知功能而可能使蛋白质失活(或激活)的位点。裂解序列的长度例如为4个残基(即P1-P4位)以匹配扩大的蛋白酶底物特异性，但是也可以更长或更短。例如，补体因子C2中裂解序列的P4-P1氨基酸残基是SLGR(SEQ IDNO：431)，但是也可以是P4-P2’序列SLGRKI(SEQ ID NO：479)，其中在P1与P1’位置(即R/K之间)位置发生裂解。因此，裂解序列内的任一或多个残基，包括P4-P2’残基包括P4-P1的任一或多个，均可以导入蛋白酶捕获多肽中，如导入丝氨酸蛋白酶抑制蛋白的RSL中以产生突变蛋白酶捕获多肽。
可以通过本领域已知的任何方法在靶底物中鉴别裂解序列(见例如公布的美国申请US2004/0146938)。在一个实例中，靶底物的裂解通过将靶底物与已知裂解所述底物的任何蛋白酶一起保温而确定。在与所述蛋白酶保温后，可以通过SDS-PAGE分离靶蛋白，并通过用蛋白质染料如考马斯亮蓝对蛋白质染色而鉴别降解产物。可以对蛋白酶解片段进行测序以确定裂解序列的性质，例如6氨基酸P4-P2’裂解序列，及特别是4氨基酸P4-P1裂解序列残基。表6鉴别了举例的靶底物相应于P4-P1位置的裂解序列。
因此，对于靶底物的任何希望的或预定的裂解序列，可以对丝氨酸蛋白酶抑制蛋白的RSL或者其它蛋白酶捕获剂中类似序列进行修饰。在一个实例中，选择的裂解序列可以是t-PA的特别有效的裂解序列。这种裂解序列是例如PFGRS(SEQ ID NO：389；见例如Ding et al.(1995)PNAS，92：7627-7631)。因此，例如可以选择具有改变的底物特异性的蛋白酶，以复制t-PA的底物特异性。由于t-PA是治疗纤维蛋白溶解疾病的常用的治疗剂，因此这种被选择的蛋白酶可以优化为t-PA的替代治疗剂，使得通常与t-PA治疗剂相关的不希望的副作用(即过度出血)最小化。
在另一实例中，可以使用本发明提供的方法将补体蛋白的裂解序列靶向为预定或希望的裂解序列以选择蛋白酶。被选择的蛋白酶对于任一或多种补体蛋白具有增加的底物特异性，其将是治疗疾病或病症的候选治疗剂，所述疾病或病症与炎症相关，例如但非限于自身免疫疾病如类风湿性关节炎和狼疮，心脏病，及其它炎症疾病如败血症和缺血-再灌注损伤(见例如临时申请系列号60/729,817)。实施例6至实施例15例证了根据通过用C2补体蛋白的裂解序列(即SLGRKI，SEQ ID NO：479)置换天然P4-P2’残基IAGRSL(SEQ ID NO：478)修饰的AT3丝氨酸蛋白酶抑制蛋白分子来选择MT-SP1蛋白酶。在任何蛋白酶捕获多肽如任何丝氨酸蛋白酶抑制蛋白多肽中可以用SLGRKI序列或者如下述的其中间体修饰或置换氨基酸残基以选择希望的任何候选蛋白酶。
在另一个实例中，裂解序列可以在VEGFR中选择，如在VEGFR的柄区域中选择，由此VEGFR在由对于裂解序列具有特异性的蛋白酶裂解时失活。VEGFR中裂解序列的例子在本文描述，及见于相关公布的美国申请系列号US20060024289和US20060002916所述。例如，丝氨酸蛋白酶抑制蛋白的RSL或者在其它蛋白酶捕获剂中的类似序列如α-2巨球蛋白中的“诱饵”区域可以被修饰为在P4-P2’的任一或多个氨基酸位置由VEGFR的裂解序列置换。在一个实例中，天然丝氨酸蛋白酶抑制蛋白中的氨基酸残基可以被修饰为含有相应于RRVR(SEQ ID NO：489)裂解序列的P4-P1位置或者全部的P4-P2’序列RRVRKE(SEQ ID NO：498)。针对这种修饰的丝氨酸蛋白酶抑制蛋白而选择的蛋白酶将是治疗VEGFR介导的疾病例如血管生成疾病的候选治疗剂。
在一些情况中，在本发明提供的方法中，丝氨酸蛋白酶抑制蛋白RSL的任一或多个P4-P2’位置或者其它蛋白酶捕获剂中的相似序列中的修饰可以在连续轮中产生以优化选择具有希望的或预定的底物特异性的蛋白酶。例如，u-PA和t-PA蛋白酶均优选在P2位的小氨基酸及在P3和P4位非常不同的氨基酸。因此，可以产生修饰的丝氨酸蛋白酶抑制蛋白，其是最终的靶裂解序列的中间体，其中第一个中间体是通过仅修饰P3和P4位置产生的以选择呈现P3和P4位置特异性的蛋白酶。然后可以将被选择的蛋白酶用作模板产生针对另外对P2位置进行改变的新丝氨酸蛋白酶抑制蛋白分子的新组合文库。
因此，例如在选择对于VEGFR裂解序列RRVR呈现增加的底物特异性的u-PA蛋白酶或其变体中，第一轮选择可以是针对中间体修饰的蛋白酶捕获多肽如丝氨酸蛋白酶抑制蛋白进行，其中仅P3和P4位置与天然序列的这些位置相比被改变。例如，在丝氨酸蛋白酶抑制蛋白PAI-1的RSL环中天然P4-P1’氨基酸是VSARM的情况中，可以通过仅置换P4和P3VEGFR裂解序列产生修饰的中间体PAI-1以产生在P4-P1’位含有RRARM(SEQ IDNO：379)的中间体丝氨酸蛋白酶抑制蛋白分子。随后轮的蛋白酶选择可以针对另外在P2位置被修饰的PAI-1丝氨酸蛋白酶抑制蛋白进行。
可以使用本领域已知的任何修饰蛋白质的方法对蛋白酶捕获剂包括丝氨酸蛋白酶抑制蛋白进行修饰。这种方法包括定点诱变，包括单位点或多位点定点诱变。同样，可以使用本领域表达与纯化多肽的标准方法对蛋白酶捕获多肽包括变体蛋白酶捕获多肽进行表达与纯化。任何宿主细胞系统均可用于表达，包括但非限于哺乳动物细胞、细菌细胞或者昆虫细胞。另外，蛋白酶捕获多肽可以进一步被修饰为包括有助于所述蛋白酶捕获多肽的鉴别与纯化的其它序列。例如，可以加入表位标记例如但非限于His标记或Flag标记以有助于所述多肽的亲和纯化。在一些实例中，蛋白酶捕获多肽被直接生物素化以有助于捕获和/或纯化。生物素化蛋白酶捕获多肽的方法在实施例16中举例说明。
如丝氨酸蛋白酶抑制蛋白分子或其它蛋白酶捕获剂的生物功能测定等测定法为本领域所已知，可用于评估在本发明的方法中修饰的蛋白酶捕获剂作为抑制剂的活性。这种测定法依赖于被修饰而用于本发明方法中的蛋白酶捕获多肽。对于PAI-1的这种测定法的例子包括例如针对标准胰蛋白酶的活性位点滴定或者标准胰蛋白酶的滴定，如实施例1举例说明。这种测定方法的另一例子是本领域已知的蛋白酶抑制测定，其中蛋白酶捕获剂抑制活性蛋白酶裂解荧光底物的能力用作蛋白酶捕获剂活性的读出结果。蛋白酶抑制测定法的例子是matriptase(MT-SP1)抑制测定法。在这种测定法的一个实例中，蛋白酶捕获剂是丝氨酸蛋白酶抑制蛋白。在特定的实施例中，丝氨酸蛋白酶抑制蛋白是AT3或者根据本发明提供的方法产生的变体AT3蛋白，所述荧光底物是RQAR-ACC。例如实施例14A举例，测量了底物的裂解。凝血酶抑制测定也可用于评估AT3或者修饰的AT3的活性。根据与蛋白酶捕获多肽或其变体通常相互作用的关联蛋白酶，类似的测定法可以被设计或者为本领域技术人员所已知。另外，预期且通常是这样的情况，即修饰的蛋白酶捕获多肽与野生型蛋白酶捕获多肽相比在正常的蛋白酶捕获活性或功能测定中具有降低的活性。
D.蛋白酶
在本发明提供的方法中，选择典型地对于预定或希望的底物呈现改变的底物特异性的候选蛋白酶。将蛋白酶、突变体蛋白酶或者其催化活性部分的集合与蛋白酶捕获多肽如本文提供的任何蛋白酶捕获多肽包括但不限于丝氨酸蛋白酶抑制蛋白或修饰的丝氨酸蛋白酶抑制蛋白接触以选择具有改变的底物特异性的蛋白酶。所述蛋白酶集合可以在固体支持物上或者于均质混合物如溶液或悬浮液中提供。被选择的蛋白酶可以作为与蛋白酶捕获多肽的稳定复合物而分离及被鉴别。被选择的蛋白酶对于希望的或预定的靶底物展示增加的催化效率和反应性，因此在例如涉及所述靶底物的任何疾病或病症中用作治疗剂的候选物。
1.候选蛋白酶
在本发明提供的方法中，选择对于疾病或病症中涉及对于希望的底物具有改变的和/或增加的特异性的蛋白酶。通常，蛋白酶是高度特异性的蛋白质，其水解靶底物同时保持其它物质完整。对于天然底物的裂解，蛋白酶呈现高度的选择性，由此有利于底物裂解而不利于非底物裂解(Coombset al.(1996)J.Biol.Chem.，271：4461-4467)。对于希望的靶底物具有改变的特异性和选择性的蛋白酶的选择使得可以使用蛋白酶作为治疗剂以选择性激活或失活蛋白酶从而减少、改善或者预防疾病或病症。本发明提供的蛋白酶捕获选择方法中使用的靶蛋白酶可以是能使与所述蛋白酶捕获剂相互作用的肽键水解的任何已知类别的蛋白酶。典型地，对于丝氨酸蛋白酶抑制蛋白而言，这种蛋白酶通常是丝氨酸或半胱氨酸蛋白酶，丝氨酸蛋白酶抑制蛋白与其反应形成共价中间体复合物。丝氨酸和半胱氨酸蛋白酶的例子是下表7列出的任何蛋白酶。典型地，本发明提供的方法中使用修饰的蛋白酶的文库以选择蛋白酶变体，所述蛋白酶变体对于靶蛋白酶捕获剂或其变体如丝氨酸蛋白酶抑制蛋白或其变体呈现增加的特异性或选择性。
本文描述了本发明提供的选择方法中可以使用和/或为了用在所述方法中而修饰的蛋白酶的例子，包括包括其催化活性部分的截短的多肽。候选蛋白酶的实例列于表7并在本文中描述(见例如www.merops.sanger.ac.uk)。所述表中描述了每个举例的候选蛋白酶的核苷酸序列及编码的氨基酸前体序列的序列标识符(SEQ ID NO)。相应于信号肽或者前肽序列以产生成熟蛋白质的编码的氨基酸也在该表中示出。另外，也示出了指明蛋白酶结构域的氨基酸(即肽酶单位)，其是组成例如各个蛋白酶催化三联体的活性位点残基。由于相互作用是动态的，因此标示的氨基酸位置只供参考和举例说明。标示的位置反映出可变化2、3、4、5或更多个氨基酸的基因座范围。在等位基因变体和物种变体之中也存在变化。本领域技术人员可以通过目测对比或者其它对比方法包括易于利用的算法和软件鉴别相应序列。
用于选择的候选蛋白酶典型是野生型蛋白酶的野生型或者修饰的或者变体形式，或者其催化活性部分，包括任一蛋白质的等位基因变体和同工型。候选蛋白酶可以通过本领域已知的任何方法产生和分离，包括从天然来源中分离、在细胞、组织或生物体中重组产生的蛋白质的分离，及通过重组方法和通过包括在芯片(in silico)步骤的方法、合成方法及本领域技术人员已知的任何方法。用于选择的候选蛋白酶的修饰可以通过本领域技术人员已知的任何方法进行，例如下文描述的任何方法。
表7：候选蛋白酶实例
a.蛋白酶类别
蛋白酶(也称作肽酶)是降解蛋白质的酶，其识别靶蛋白质内的氨基酸序列或者多肽底物。在识别底物氨基酸序列时，蛋白酶催化靶蛋白内肽键的水解或裂解。依赖于所述肽键在靶序列的全长序列内的位置，靶蛋白的这种水解可以失活或者在一些情况中激活靶位。
蛋白酶基于其攻击蛋白的方式而分为外切蛋白酶和内切蛋白酶类别。蛋白酶或内肽酶攻击蛋白质内部产生大肽。肽酶或者外肽酶攻击蛋白质的末端或片段产生小肽和氨基酸。肽酶根据其作用模式而可分为如下类别：氨基肽酶，从氨基末端裂解氨基酸；羧肽酶，从羧基末端裂解氨基酸；二肽基肽酶，裂解两个氨基酸；二肽酶，裂解二肽；三肽酶，裂解三肽的氨基酸。大多数蛋白酶较小，为21000至45000道尔顿。许多蛋白酶是以称作酶原的失活形式合成和分泌，随后通过蛋白酶解而激活。这样改变了所述酶活性位点的结构。
蛋白酶利用一些独特类型的催化机制(Barret et al.(1994)Meth.Enzymol.244：18-61；Barret et al.(1994)Meth.Enzymol 244：461-486；Barret etal.(1994)Meth.Enzymol.248：105-120；Barret et al.(1994)Meth.Enzymol.248：183-228)。基于其催化机制，羧肽酶被再分为丝氨酸-、金属-和半胱氨酸型羧肽酶，内肽酶被再分为丝氨酸-、半胱氨酸-、天冬氨酸-、苏氨酸-和金属内肽酶。丝氨酸肽酶活性中心包含丝氨酸残基，天冬氨酸肽酶催化中心内具有两个天冬氨酸，半胱氨酸型肽酶具有半胱氨酸残基，苏氨酸型肽酶具有苏氨酸残基，金属肽酶在催化机制中使用金属离子。通常，蛋白酶可以基于其催化活性而分类，由此蛋白酶的类别可包括丝氨酸蛋白酶、半胱氨酸蛋白酶、天冬氨酸蛋白酶、苏氨酸蛋白酶或金属蛋白酶。蛋白酶的催化活性为裂解靶底物所必需。因此，改变蛋白酶的催化活性的修饰(即修饰特异性/选择性)可影响蛋白酶裂解特定底物的能力。
每种蛋白酶均具有一系列沿着活性位点口袋排列并且使其与底物直接接触的氨基酸。肽酶的晶体结构示出所述活性位点通常位于相邻结构域之间分子表面上的沟中，底物特异性是由沿着催化位点的一或两侧的沟排列的结合位点的性质决定，所述催化位点与易断裂键的水解相关。因此，肽酶的特异性由每个亚位点容纳单氨基酸残基的侧链的能力加以描述。所述位点从催化位点编号，S1，S2...Sn朝向底物的N末端，S1′，S2′...Sn′朝向C末端。其容纳的残基分别编号为P1，P2...Pn和P1′，P2′...Pn′。靶蛋白的裂解在P1与P1’之间被催化，其中从所述多肽底物的N末端至C末端的氨基酸残基被标为(Pi，...，P3，P2，P1，P1′，P2′，P3′，...，Pj)，蛋白酶上其相应的结合识别口袋被标为(Si，...，S3，S2，S1，S1′，S2′，S3′，...，Sj)(Schecter and Berger(1967)Biochem Biophys Res Commun 27：157-162)。因此，P2与S2相互作用，P1与S1相互作用，P1’与S1’相互作用等。结果，蛋白酶的底物特异性来自活性位点中的S1-S4位，在此蛋白酶与肽底物序列的P1-P4残基接触。在一些情况中，活性位点的S1-S4结合口袋之间的相互作用很小(若有的话)，由此每个结合口袋看起来识别并结合独立于其它结合口袋的肽底物序列上的相应残基。因此，可以改变一个口袋中的特异性决定簇，而不影响其它口袋的特异性。基于家族成员的众多结构与建模，针对许多蛋白酶已经描述了导致扩大的底物特异性及与底物的其它二级相互作用的表面残基，所述蛋白酶包括丝氨酸蛋白酶、半胱氨酸蛋白酶、天冬氨酸蛋白酶、金属蛋白酶和苏氨酸蛋白酶家族(见例如Wang et al.，(2001)Biochemistry 40(34)：10038-46；Hopfner et al.，(1999)Structure Fold Des.7(8)：989-96；Friedrich et al.(2002)J Biol Chem.277(3)：2160-8；Waugh et al.，(2000)Nat Struct Biol.7(9)：762-5；Cameron et al.，(1993)J Biol Chem.268：11711；Cameron et al.，(1994)J Biol Chem.269：11170)。
i.丝氨酸蛋白酶
丝氨酸蛋白酶(SP)包括分泌的酶及隐蔽在胞质贮存细胞器中的酶，它们具有多种生理作用，包括参与血液凝固、创伤愈合、消化、免疫应答及肿瘤侵入和转移。例如，胰凝乳蛋白酶、胰蛋白酶和弹性蛋白酶在消化道中起作用；因子10、因子11、凝血酶和纤溶酶参与凝血及创伤愈合；Clr、Cls和C3转变酶在补体激活中起作用。
称作II型跨膜丝氨酸蛋白酶的一类细胞表面蛋白是具有胞外结构域的膜锚定蛋白质的蛋白酶。作为细胞表面蛋白，其在胞内信号转导和介导细胞表面蛋白酶解事件中起作用。其它丝氨酸蛋白酶是膜结合蛋白酶，以相似方式起作用。其它蛋白酶是分泌的蛋白酶。许多丝氨酸蛋白酶在结合细胞表面受体时发挥其活性，并因此作用于细胞表面。细胞表面蛋白酶解是产生介导多种细胞功能的生物学活性蛋白质的机制。
丝氨酸蛋白酶，包括分泌的和跨膜丝氨酸蛋白酶，参与包括肿瘤发生和进展的一些过程。虽然这些蛋白酶的精确作用还未完全阐释，但是丝氨酸蛋白酶及其抑制剂参与许多胞内和胞外生理过程的控制，包括癌细胞侵入和转移扩散中的退化作用及肿瘤进展中涉及的肿瘤新血管生成。蛋白酶参与胞外基质(ECM)的降解和重塑，并导致组织重塑，且为癌症侵入和转移所必需。一些蛋白酶的活性和/或表达已经示出与肿瘤进展和发生相关。
已经鉴别了超过20个家族的丝氨酸蛋白酶(称作S1-S27)，基于结构相似性及其它功能证据将这些家族分为6个异种集团(SA、SB、SC、SE、SF和SG)(Rawlings ND et al.(1994)Meth.Enzymol.244：19-61)。一些丝氨酸肽酶的反应机制中存在相似性。胰凝乳蛋白酶、枯草杆菌蛋白酶和羧肽酶C异种集团具有丝氨酸、天冬氨酸和组氨酸的催化三联体：丝氨酸作为亲核体，天冬氨酸作为亲电体，组氨酸作为碱。催化残基的几何方向在家族之间是相似的，尽管存在不同的蛋白质折叠。催化残基的线性排列通常反映异种集团的关系。例如，胰凝乳蛋白酶异种集团(SA)中的催化三联体的顺序是HDS，但是在枯草杆菌蛋白酶异种集团(SB)中的顺序是DHS，在羧肽酶异种集团(SC)中的顺序是SDH。
胰凝乳蛋白酶超家族的丝氨酸蛋白酶的例子包括组织型纤溶酶原激活物(tPA)、胰蛋白酶、胰蛋白酶样蛋白酶、胰凝乳蛋白酶、纤溶酶、弹性蛋白酶、尿激酶(或者尿型纤溶酶原激活物，u-PA)、顶体蛋白、激活的C蛋白、C1酯酶、组织蛋白酶G、类胰凝乳蛋白酶(chymase)，及血液凝固级联中的蛋白酶包括激肽释放酶、凝血酶和因子VIIa、IXa、Xa、XIa和XIIa(Barret，A.J.，In：Proteinase Inhibitors，Ed.Barrett，A.J.，Et al.，Elsevier，Amsterdam，Pages 3-22(1986)；Strassburger，W.et al.，(1983)FEBS Lett.，157：219-223；Dayhoff，M.O.，Atlas of Protein Sequence and Structure，Vol 5，NationalBiomedical Research Foundation，Silver Spring，Md.(1972)；和Rosenberg，R.D.et al.(1986)Hosp.Prac.，21：131-137)。
丝氨酸蛋白酶家族中蛋白酶活性依赖于形成其活性位点的一组氨基酸残基。其中一个残基总是丝氨酸，因此其名称为丝氨酸蛋白酶。例如，胰凝乳蛋白酶、胰蛋白酶和弹性蛋白酶共有一个相似结构，且所有这三种蛋白酶的活性丝氨酸残基位于相同位置(Ser-195)。尽管存在相似性，但它们具有不同的底物特异性，它们在蛋白质消化期间裂解不同的肽键。例如，胰凝乳蛋白酶优选羰基碳是被裂解肽键一部分的残基的芳族侧链。胰蛋白酶优选在这个位置正电荷的Lys或Arg。丝氨酸蛋白酶在其底物识别性质方面显著不同：一些是高特异性的(即所述蛋白酶参与血液凝固和免疫补体系统)；一些仅是部分特异性的(即哺乳动物消化蛋白酶胰蛋白酶和胰凝乳蛋白酶)；其它如枯草杆菌蛋白酶是细菌蛋白酶，是完全非特异性的。尽管在特异性方面存在这些差异，但是丝氨酸蛋白酶的催化机制是很保守的。
丝氨酸蛋白酶裂解靶蛋白的机制基于丝氨酸对靶定肽键的亲核攻击。半胱氨酸、苏氨酸或者与天冬氨酸或金属缔合的水分子也可以发挥这种作用。在许多情况中，所述基团的亲核性质通过组氨酸的存在而改良，通过天冬氨酸保持“质子受体状态”。排列的丝氨酸、组氨酸和天冬氨酸的侧链构成大多数丝氨酸蛋白酶共有的催化三联体。例如，胰凝乳蛋白酶及与胰凝乳蛋白酶是相同家族成员的丝氨酸蛋白酶如MTSP-1的活性位点残基是Asp102、His57和Ser195。
胰凝乳蛋白酶超家族的所有丝氨酸蛋白酶的催化结构域既具有序列同源性也具有结构同源性。所述序列同源性包括如下保守性：1)特征性活性位点残基(例如在胰蛋白酶情况中的Ser195、His57和Asp102)；2)氧负离子洞(oxyanion hole)(例如在胰蛋白酶情况中的Gly193、Asp194)；及3)形成所述结构中二硫键的半胱氨酸残基(Hartley，B.S.，(1974)Symp.Soc.Gen.Microbiol.，24：152-182)。所述结构同源性包括：1)特征在于两个希腊钥匙结构的共同折叠(Richardson，J.(1981)Adv.Prot.Chem.，34：167-339)；2)催化残基的共同排列；及3)分子核心内结构的细节保存(Stroud，R.M.(1974)Sci.Am.，231：24-88)。
在丝氨酸蛋白酶的胰凝乳蛋白酶家族中，底物与酶之间的主链相互作用是完全保守的，但是侧链相互作用显著不同。含有活性位点的S1-S4口袋的氨基酸特性决定了该特定口袋的底物特异性。将一个丝氨酸蛋白酶的氨基酸移植至相同折叠的另一蛋白酶，使得一种蛋白酶的特异性被修饰为另一种的特异性。典型地，含有S1-S4口袋的蛋白酶的氨基酸是具有侧链的那些氨基酸，所述侧链距底物4-5埃。这些氨基酸与蛋白酶底物的相互作用通常称作“第一层(first shell)”相互作用，因为其直接接触底物。然而，还可以具有最终定位第一层氨基酸的“第二层”和“第三层”相互作用。第一层和第二层底物结合作用主要由β-桶结构域之间的环决定。由于这些环不是蛋白质的核心元件，因此折叠的完整性被保持，同时在分子水平实现必需的代谢或调节微环境(niches)的进化期间可以选择具有新的底物特异性的环变体。典型地，对于丝氨酸蛋白酶而言，一级序列中的如下氨基酸是特异性决定簇(基于胰凝乳蛋白酶编号)：195，102，57(催化三联体)；189，190，191，192和226(S1)；57，58与64之间的环及99(S2)；192，217，218(S3)；Cys168与Cys180之间的环、215及97至100(S4)；及41和151(S2’)，其中S1位置中的氨基酸影响P1特异性，S2位置中的氨基酸影响P2特异性，S3位置中的氨基酸影响P3特异性，S4位置中的氨基酸影响P4特异性。丝氨酸蛋白酶中的第189位是埋入所述口袋底部的残基，决定S1特异性。各种丝氨酸蛋白酶的结构决定簇列于表8中，基于成熟胰凝乳蛋白酶编号方式进行编号，每个指定蛋白酶的蛋白酶结构域与胰凝乳蛋白酶的蛋白酶结构域排列对比。在Cys168-Cys182和60’s环各列标题下面的数字表示这两个氨基酸之间环中及所述环中氨基酸数目。在Cys191-Cys220一列标题下的是/否表示蛋白酶中是否存在二硫键。这些区域在胰凝乳蛋白酶样丝氨酸蛋白酶家族中可有所不同，表示其自身的结构决定簇。改变S1-S4口袋中任一或多个氨基酸的蛋白酶修饰影响蛋白酶对于靶底物的特异性或选择性。
表8：各种丝氨酸蛋白酶的结构决定簇
(a)尿激酶型纤溶酶原激活物(u-PA)
尿激酶型纤溶酶原激活物(u-PA，也称作尿纤溶酶原激活物)是用作本发明方法中用于选择的候选物的举例蛋白酶。u-PA以SEQ ID NO：190示出，其编码SEQ ID NO：191所示前体氨基酸序列。u-PA在尿液、血液、精液及在许多癌组织中发现。其参与多种生物学过程，与纤溶酶原转变为纤溶酶相关，其自身是一种丝氨酸蛋白酶。纤溶酶通过裂解多种分子包括纤维蛋白、纤连蛋白、蛋白聚糖和层粘连蛋白而在许多正常和病理学过程中起作用，包括例如细胞迁移和组织破坏。u-PA在伤口愈合、炎症细胞迁移、新血管生成和肿瘤细胞侵润期间参与组织重塑。u-PA也裂解及激活其它底物，包括但不限于肝细胞生长因子/分散因子(scatter factor)(HGF/SF)、膜型1基质金属蛋白酶的潜伏形式(MT-SP1)及其它底物。
u-PA的成熟形式是411个残基的蛋白质(相应于SEQ ID NO：191所示含有20个氨基酸信号肽的前体形式的氨基酸序列中第21-431位氨基酸)。u-PA含有三个结构域：丝氨酸蛋白酶结构域、kringle结构域及生长因子结构域。在成熟形式的人u-PA中，第1-158位氨基酸表示N末端A链，包括生长因子结构域(第1-49位氨基酸)、kringle结构域(第50-131位氨基酸)及结构域间接头区(第132-158位氨基酸)。第159-411位氨基酸表示C末端丝氨酸蛋白酶结构域或B链。u-PA作为单链酶原分子形式合成及分泌，其由多种蛋白酶转变为活性双链u-PA，所述蛋白酶包括例如纤溶酶、激肽释放酶、组织蛋白酶B和神经生长因子-γ。在成熟u-PA序列的第158与159位残基(相应于SEQ ID NO：191所示第178与179位氨基酸残基)之间裂解为双链形式。获得的两个链通过二硫键连接在一起，形成双链形式的u-PA。
u-PA通过与高亲和性细胞表面受体uPAR结合而被调节。u-PA与uPAR的结合增加了纤溶酶原激活速度及增强了胞外基质降解和细胞侵入。在uPAR与u-PA之间形成的二元复合物与膜结合纤溶酶原相互作用形成更高级的激活复合物，降低纤溶酶原激活的Km(即对于底物的近似亲和性的动力学速率常数)(Bass et al.(2002)Biochem.Soc，Trans.，30：189-194)。另外，u-PA与uPAR的结合保护蛋白酶免于关联抑制剂即PAI-1的抑制。这是因为通常存在于血浆中的单链u-PA对于PAI-1抑制不敏感，血浆中任何激活的u-PA将被PAI-1抑制。与受体结合的激活的u-PA可由PAI-1充分抑制，然而PAI-1不能接近结合的活性分子(Bass et al.(2002)Biochem.Soc，Trans.，30：189-194)。结果，u-PA主要在细胞表面起作用且其功能与纤溶酶依赖性细胞外周蛋白酶解的激活相关。
u-PA与t-PA(在下文讨论)的扩大的底物特异性相似，这是由于这二者均可以将纤溶酶原裂解为活性纤溶酶。u-PA和t-PA二者对于在P1Arg之后裂解均具有高特异性，且相似地示出优选在P2位置的小氨基酸。P3和P4这两个位置均是u-PA和t-PA的底物特异性决定簇，P3位置的作用特别显著(Ke et al.(1997)J.Biol.Chem.，272：16603-16609)。对于在P3位置氨基酸的偏爱是独特的，且是两个蛋白酶之间改变的底物分辨性的主要决定簇。t-PA对于在P3位置的芳族氨基酸(Phe和Tyr)具有偏爱，而u-PA对于小极性氨基酸(Thr和Ser)具有偏爱(见例如Ke et al.(1997)J.Biol.Chem.，272：16603-16609；Harris et al.(2000)PNAS，97：7754-7759)。
(b)组织纤溶酶原激活物(t-PA)
在本发明的方法中针对蛋白酶捕获剂选择候选蛋白酶也包括例如丝氨酸蛋白酶组织纤溶酶原激活物(t-PA)及其变体。t-PA是一种丝氨酸蛋白酶，其将纤溶酶原转变为纤溶酶，参与纤维蛋白溶解或者血凝块形成。重组t-PA在特征在于血液凝固例如脑中风的疾病中用作治疗剂。t-PA基因的可变剪接产生三个转录物。主要的转录物以SEQ ID NO：192示出，其编码SEQ IDNO：193所示含有20-23个氨基酸信号序列和12-15个氨基酸前序列的前体蛋白质。其它转录物以SEQ ID NO：194和196示出，其分别编码具有SEQ IDNO：195和197所示氨基酸序列的前体蛋白。没有信号序列和前肽序列的t-PA成熟序列是527个氨基酸。
t-PA由血管内皮分泌并且以单链形式在血液中循环。与许多其它丝氨酸蛋白酶不同，单链或“酶原”形式的t-PA具有高催化效率。在存在纤维蛋白的条件下t-PA的活性增加。在不存在纤维蛋白的条件下，单链t-PA的活性是双链t-PA活性的大约8％，然而在存在纤维蛋白的条件下，单链和双链形式的t-PA显示相似活性(Strandberg et al.(1995)J.Biol.Chem.，270：23444-23449)。因此，单链t-PA的激活可以通过激活裂解(即酶原裂解)产生双链形式或者通过结合辅因子纤维蛋白而实现。在由纤溶酶、组织激肽释放酶和激活的因子X在氨基酸位置Arg275-Ile276(相应于SEQ ID NO：193所示氨基酸序列中Arg310-Ile311)裂解后发生激活裂解，导致活性双链形式t-PA产生。双链多肽含有A和B链，其通过链间二硫键连接。
成熟t-PA含有16个二硫键，组成5个独特结构域(Gething et al.(1988)，The EMBO J.，7：2731-2740)。成熟蛋白的第4-50位残基形成指状结构域，第51-87位残基形成EGF样结构域，第88-175和176-263位残基形成两个kringle结构域，每个kringle结构域均含有三个结构域内二硫键，成熟分子的第277-527位残基(相应于SEQ ID NO：193所示前体序列的第311-562位氨基酸残基)组成丝氨酸蛋白酶结构域。
与作为细胞受体结合激活物的u-PA相反，t-PA发挥纤维蛋白依赖性循环激活酶作用。同样，单链和双链形式的t-PA对于其关联抑制剂例如PAI-1的抑制作用均敏感，但是PAI-1对双链t-PA的抑制比对单链t-PA的抑制快大约1.4倍(Tachias et al.(1997)J Biol.Chem.，272：14580-5)。t-PA通过结合内皮细胞上的细胞结合位点Annexin-II可以被保护免于被抑制。因此，尽管t-PA和u-PA均裂解并激活纤溶酶原，但是t-PA在血液中的作用支持t-PA是纤溶酶原的主要纤维蛋白溶解性激活物，而u-PA是纤溶酶原的主要细胞激活物。
(c)MT-SP1
膜型丝氨酸蛋白酶MT-SP1(也称作matriptase，TADG-15，肿瘤发生能力阻抑物14，ST14)是在本发明提供的方法中用于选择的蛋白酶以选择对于希望的或预定的底物裂解序列具有改变的底物特异性的变体的例子。MT-SP1的序列示于SEQ ID NO：252，其编码具有SEQ ID NO：253所示氨基酸序列的855个氨基酸的多肽。其是具有C末端丝氨酸蛋白酶结构域的多结构域蛋白酶(Friedrich et al.(2002)J Biol Chem 277(3)：2160)。已经分离了683个氨基酸的蛋白酶变体，但是这个蛋白质看起来是截短形式或胞外结构域形式。
MT-SP1在前列腺、乳腺和结肠直肠癌中高表达或活性，其可在乳腺癌和前列腺癌转移中起作用。MT-SP1也在多种上皮组织中表达，在人胃肠道和前列腺中具有高水平活性和/或表达。已知其它物种的MT-SP1。例如，已经鉴别了MT-SP1小鼠同系物，将其称作epithin。
MT-SP1含有一个跨膜结构域，两个CUB结构域，四个LDLR重复及在SEQ ID NO：253所示序列的第615-854位氨基酸(或者根据文献报道不同在第615-855位氨基酸)之间的一个丝氨酸蛋白酶结构域(或肽酶S1结构域；也称作B-链)。所述蛋白酶结构域的氨基酸序列示于SEQ ID NO：505，由SEQID NO：504所示的核酸序列编码。MT-SP1作为酶原被合成，通过裂解被激活为双链形式。另外，单独的单链蛋白酶解结构域是催化活性和功能性的。
称作CB469的MT-SP1变体具有相应于SEQ ID NO：253或505所示MT-SP1野生型序列的C122S突变(基于胰凝乳蛋白酶编号)，其呈现在噬菌粒载体上改良的展示。这种变体MT-SP1示于SEQ ID NO：515(全长MT-SP1)或者SEQ ID NO：507(蛋白酶结构域)，可用于下文所述的方法中。
MT-SP1属于丝氨酸蛋白酶的肽酶S1家族(也称作胰凝乳蛋白酶家族)，其也包括胰凝乳蛋白酶和胰蛋白酶。通常，胰凝乳蛋白酶家族成员与胰凝乳蛋白酶呈现序列和结构同源性。MT-SP1在本文根据成熟胰凝乳蛋白酶编号，其蛋白酶结构域与胰凝乳蛋白酶的蛋白酶结构域进行对比并因此对其残基进行编号。基于胰凝乳蛋白酶编号，活性位点残基是Asp102、His57和Ser195(相应于SEQ ID NO：253中Asp711、His656和Ser805)。线性氨基酸序列可以与胰凝乳蛋白酶的序列对比并根据胰凝乳蛋白酶的β折叠编号。在β折叠之间的环中发生插入和缺失，但是在整个结构家族中，核心折叠是保守的。所述丝氨酸蛋白酶与底物以保守的β折叠方式相互作用。在底物与酶之间可发生直至6个保守氢键。胰凝乳蛋白酶家族的所有丝氨酸蛋白酶在其为催化活性所必需的蛋白酶结构域的N末端均具有一个保守区域(即IIGG、VVGG或IVGG，其中这个四联体中第一个氨基酸根据胰凝乳蛋白酶编号方式编号，称作Ile16。这种编号不反映前体序列的长度)。
已经使用位置扫描合成的组合文库和底物噬菌体展示方法对蛋白酶结构域中MT-SP1的底物特异性进行作图(Takeuchi et al.(2000)J Biol Chem275：26333)。底物中由MT-SP1识别的裂解残基含有在P4位的Arg/Lys和在P3位的碱性残基或Gln、在P2位的小残基、在P1位的Arg或Lys及在P1’位的Ala。有效的底物在P4至P1位点分别含有Lys-Arg-Ser-Arg。通常，对于MT-SP1的底物特异性揭示了这样的倾向，即如果P3是碱性的，则P4趋于是非碱性的；如果P4是碱性的，则P3趋于是非碱性的。已知的MT-SP1底物包括例如蛋白酶激活的受体-2(PAR-2)、单链uPA(sc-uPA)、MT-SP1的原形式(proform)及肝细胞生长因子(HGF)符合MT-SP1特异性底物的裂解序列。
MT-SP1可以与胰蛋白酶同样有效地裂解选择的合成底物。但是比胰蛋白酶呈现更受限的底物特异性。MT-SP1的催化结构域具有胰蛋白酶样丝氨酸蛋白酶的全部结构折叠，但是展示独特的性质如疏水性/酸性S2/S4亚位点和暴露的60环。相似地，MT-SP1在可及的Lys或Arg残基不是不加选择地裂解肽底物，而是需要识别易断裂肽键周围的其它残基。这种对于扩大的一级序列的要求突出了MT-SP1对其底物的特异性。例如，尽管MT-SP1裂解蛋白酶激活受体-2(PAR-2)(展示P4至P1靶序列Ser-Lys-Gly-Arg)，但是所述酶不激活与这个底物密切关联的蛋白质，如PAR-1、PAR-3和PAR-4，这些蛋白质未展示易断裂键附近与扩大的MT-SP1特异性匹配的靶序列(见Friedrich et al.(2002)J Biol Chem 277：2160)。
MT-SP1的蛋白酶结构域由前区域(pro-region)和催化结构域组成。所述多肽的催化活性部分自成熟蛋白第611位氨基酸残基的自身活化位点之后开始(见例如SEQ ID NO：253，在RQAR之后是残基VVGG)。MT-SP1与胰蛋白酶的S1口袋对于Lys以及Arg P1残基的良好互补性相似，从而解释了在胰蛋白酶的底物裂解中的一些相似性。P1-Lys残基的容纳由Ser190介导，其侧链提供了另外的氢键受体以稳定隐蔽的α-铵基团(见Friedrich et al.(2002)J Biol Chem 277：2160)。S2口袋适合于容纳P2氨基酸的小至中等大小的疏水性侧链，且通常接受在P2位置的多种氨基酸。在底物结合时，S2亚位点不是刚性的，这由Phe99苄基基团的旋转证实。在P3(对于Gln或者碱性残基)和P4(对于Arg或Lys残基)位置的底物氨基酸看起来由S3和S4口袋与Asp-217和/或Asp-96的酸性侧链的静电相互作用而介导，当它们接近酶活性位点裂口处时可有利地预定向(pre-orient)特定的碱性肽底物。P3残基的侧链也能与Gln192的氨甲酰基团形成氢键，或者PS侧链可伸入S4亚位点中与Phe97形成氢键，从而削弱与Gly216的主链间氢键。在任一构象中，碱性P3侧链能与MT-SP1 S4口袋的负电位有利地相互作用。相互的电荷补偿及从相同S4位点排除解释了在良好的MT-SP1底物中P3和P4位置同时出现Arg/Lys残基的可能性低。通常，造成对于底物结合的扩大的特异性的MT-SP1氨基酸位置(基于胰凝乳蛋白酶编号)包括：146和151(S1’)；189、190、191、192、216、226(S1)；57、58、59、60、61、62、63、64、99(S2)；192、217、218、146(S3)；96、97、98、99、100、168、169、170、170A、171、172、173、174、175、176、178、179、180、215、217、224(S4)。
ii.半胱氨酸蛋白酶
半胱氨酸蛋白酶具有包含半胱氨酸巯基基团的催化机制。半胱氨酸巯基由相邻的组氨酸去质子化，随后半胱氨酸对于肽羰基碳进行亲核攻击。连接新的羧基末端与半胱氨酸硫醇的硫酯是所述反应的中间体(与丝氨酸蛋白酶的酰基酶中间体相似)。半胱氨酸蛋白酶包括木瓜蛋白酶、组织蛋白酶、胱天蛋白酶和钙激活中性蛋白酶。
木瓜蛋白酶样半胱氨酸蛋白酶是与木瓜蛋白酶结构相似的硫醇依赖性内肽酶家族。其形成具有标记为R和L(代表右和左)的结构域的双结构域蛋白质及来自这两个结构域的环形成底物识别裂口。它们具有由氨基酸Cys25、His159和Asn175组成的催化三联体。与基于底物的β折叠构象识别且蛋白酶解靶肽的丝氨酸蛋白酶不同，这个蛋白酶家族不具有明确的口袋用于底物识别。主要底物识别发生在P2氨基酸(与丝氨酸蛋白酶中在P1残基相比)。
许多半胱氨酸蛋白酶(人组织蛋白酶L、V、K、S、F、B、木瓜蛋白酶和cruzain)的底物特异性已经使用完整多样位置扫描合成组合文库(complete diverse positional scanning synthetic combinatorial library，PS-SCL)确定。该完整文库含有P1、P2、P3和P4四肽底物，其中一个位置保持固定，其它三个位置随机具有20个可能氨基酸的等摩尔混合物，提供共～160000个四肽序列的多样性。
总之，P1特异性在组织蛋白酶之间几乎相同，Arg和Lys优先，且耐受小的脂族氨基酸。在P2位置发现许多选择性，其中人组织蛋白酶对于疏水性氨基酸具有严格选择性。令人感兴趣地，P2对于疏水性残基的特异性在芳族氨基酸如Phe、Tyr和Trp(组织蛋白酶L、V)与大脂族氨基酸如Val或Leu(组织蛋白酶K、S、F)之间有所不同。与P2位置相比，在P3位置的选择性严格性明显较低。然而，一些蛋白酶对于脯氨酸(组织蛋白酶V、S和木瓜蛋白酶)、亮氨酸(组织蛋白酶B)或者精氨酸(组织蛋白酶S、cruzain)具有独特的偏爱。所述蛋白酶示出在P4位置具有广谱特异性，因为无一氨基酸相对于其它氨基酸被选择。
S2口袋是最具选择性的且是蛋白酶底物识别位点的最被特征化的。其通过如下空间位置的氨基酸限定(木瓜蛋白酶编号)：66、67、68、133、157、160和205。第205位置发挥与丝氨酸蛋白酶中第189位置相似的作用，这是埋入口袋底部的决定特异性的残基.。其它特异性决定簇包括如下氨基酸(基于木瓜蛋白酶编号)：61和66(S3)；19、20和158(S1)。各种半胱氨酸蛋白酶的结构决定簇列于表9。典型地，对半胱氨酸蛋白酶例如木瓜蛋白酶进行的修饰影响蛋白酶对于靶底物包括补体蛋白靶底物的特异性或选择性，所述修饰用以改变扩大的特异性结合口袋或者相互作用的其它二级位点中的任一或多个氨基酸。
表9：各种半胱氨酸蛋白酶的结构决定簇
E.用于筛选的修饰的蛋白酶及集合
蛋白酶或其变体可用于本发明的方法中以鉴别具有希望的底物特异性的蛋白酶，通常所述底物特异性被改变、改良或优化。本发明提供的方法中使用的修饰的蛋白酶可以通过使用本领域已知的任何方法突变蛋白酶的任一或多个氨基酸残基而产生(也见公布的美国申请2004/0146938所述)。用于修饰及本发明方法的蛋白酶包括例如全长野生型蛋白酶、具有足够催化活性例如蛋白酶解底物的蛋白酶的已知变体形式或片段。这种修饰的蛋白酶可以利用靶蛋白酶捕获剂单独筛选，所述捕获剂例如是丝氨酸蛋白酶抑制蛋白或者修饰的丝氨酸蛋白酶抑制蛋白；或者这种修饰的蛋白酶可以作为集合被筛选，例如通过使用展示文库筛选，所述展示文库包括组合文库，其中蛋白酶通过例如噬菌体展示、细胞表面展示、珠展示、核糖体展示或者其它方式进行展示。由于稳定的共价抑制复合物的形成而对蛋白酶捕获剂或其修饰的形式呈现特异性和/或选择性的蛋白酶的选择可以通过本领域技术人员已知的任何检测方案进行，包括但不限于亲和标记和/或纯化法、ELISA、生色测定法、基于荧光测定法(例如荧光猝灭法或者FRET)等。
1.变体蛋白酶的产生
突变蛋白酶序列的方法的例子包括导致全部序列随机诱变的方法或者导致蛋白酶序列的选择序列或结构域发生聚焦诱变的方法。在一个实例中，对蛋白酶进行突变的数目是1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19或20个。在优选的实施方案中，所述突变赋予增加的底物特异性。在一些实例中，蛋白酶变体的活性比野生型蛋白酶活性增加至少10倍、100倍或1000倍。在相关方面，所述活性增加是底物特异性增加。
a.随机诱变
随机诱变方法包括例如使用大肠杆菌(E.coli)XL1red、UV照射、化学修饰例如脱氨、烷化或者使用碱基类似物诱变剂，或者PCR方法如DNA改组、盒诱变、定点随机诱变，或者易错PCR(见例如美国申请号：2006-0115874)。这些实例包括但不限于通过羟胺进行化学修饰(Ruan，H.，etal.(1997)Gene 188：35-39)、使用dNTP类似物(Zaccolo，M.，et al.(1996)J.Mol.Biol.255：589-603)，或者使用可商购的随机诱变试剂盒如GeneMorph基于PCR的随机诱变试剂盒(Stratagene)或者Diversify随机诱变试剂盒(Clontech)。所述Diversify随机诱变试剂盒通过改变反应混合物中锰(Mn2+)和dGTP的量从而可以选择指定DNA序列的希望的突变率(2-8个突变/1000个碱基对)。锰离子水平增加最初增加了突变率，通过增加dGTP的浓度可进一步增加突变率。通过额外进行PCR循环甚至可以达到更高的突变率。
b.聚焦诱变
聚焦诱变可以通过在基因序列的预定区域中产生一或多个突变而实现，例如在介导催化活性的蛋白酶结构域区域中产生突变。在一个实例中，使用任何标准的单位点或多位点定向诱变试剂盒例如QuikChange(Stratagene)对蛋白酶的任一或多个氨基酸进行突变。在另一个实例中，通过饱和诱变方法(Zheng et al.(2004)Nucl.Acids.Res.，32：115)对蛋白酶的任一或多个氨基酸进行突变，所述方法例如是对活性位点残基进行诱变。在这个实例中，形成蛋白酶S1-S4口袋的残基(在此所述蛋白酶与肽底物的P1-P4残基接触)和/或已经示出是特异性的重要决定簇的残基单独或组合地被突变为每一种可能的氨基酸。在一些情况中，在活性位点的S1-S4口袋之间存在少许(如果有的话)相互作用，由此每个口袋均看起来识别并结合独立于其它口袋的肽底物序列上的相应残基。因此，可以改变一个口袋中的特异性决定簇通常不影响其它口袋的特异性。在一个举例的实施方案中，使用饱和诱变技术，其中沿着口袋排列的残基被突变为20种可能氨基酸的每一种(见例如Kunkle方法所述，Current Protocols in Molecular Biology，JohnWiley and Sons，Inc.，Media Pa.)。在这种技术中，可以合成简并诱变寡核苷酸引物，其在编码选择的氨基酸的希望的密码子含有随机化的核苷酸。随机化方案的例子包括NNS-或NNK-随机化，其中N代表任何核苷酸，S代表鸟嘌呤或者胞嘧啶，K代表鸟嘌呤或胸腺嘧啶。所述简并诱变引物与单链DNA模板退火，加入DNA聚合酶以合成所述模板的互补链。在连接后，将双链DNA模板转化进大肠杆菌中以进行扩增。
本文描述了形成举例的蛋白酶的扩大的底物结合口袋的氨基酸。通常，已知蛋白酶的底物特异性，例如通过基于蛋白酶和底物复合物的三维结构的分子建模而获知(见例如Wang et al.，(2001)Biochemistry 40(34)：10038；Hopfner et al.，Structure Fold Des.19997(8)：989；Friedrich et al.，(2002)J BiolChem 277(3)：2160；Waugh et al.，(2000)Nat Struct Biol.7(9)：762)。例如，可以对MT-SP1中与底物特异性相关的任一或多个残基进行聚焦突变，所述残基位包括(基于胰凝乳蛋白酶编号)：195、102、157(催化三联体)；189、190、191、192、216和226(S1)；57、58、59、60、61、62、63、64、99(S2)；146、192、217、218(S3)；96、97、98、99、100、168、169、170、170A、171、172、173、174、175、176、178、179、180、215、217、224(S4)。在另一个实例中，木瓜蛋白酶家族蛋白酶中氨基酸残基的突变可以是影响P2特异性的任一或多个残基包括66-68、133、157、160和/或215(标准木瓜蛋白酶编号)中的突变。另外，与蛋白酶底物不直接接触、但是影响接触残基的位置和/或构象的残基(例如上文列出的那些残基)也可以被突变以改变蛋白酶支架的特异性。
在另一个实例中，用于诱变的聚焦氨基酸可以通过具有相似底物特异性的同源蛋白酶的序列对比而选择。共有氨基酸残基可以通过对比同源蛋白质的氨基酸序列而鉴别，例如对比参与底物结合的蛋白酶区域。典型地，具有相似底物特异性的蛋白酶共享共有氨基酸，例如底物结合口袋中的氨基酸在对比的蛋白酶之间可以相同或相似。此外，可以对比具有不同底物特异性的蛋白酶的氨基酸序列以鉴别可以参与底物识别的氨基酸。这些方法可以与例如三维建模方法组合用于鉴别进行诱变的靶残基。
在另一个实例中，聚焦诱变可限于在最初的蛋白酶筛选循环中鉴别为热点的氨基酸。例如，在从随机诱变的组合文库中选择蛋白酶之后，在筛选方法中一再地观测到几个“热点”位置并加以选择。最通常地，由于随机诱变广泛突变多肽序列，但是在每个位点仅极少的几个突变，因此使用聚焦诱变作为特异性靶热点位置的第二种策略以进一步诱变。热点位置的聚焦诱变可以在特定位置进行更多样和深入的诱变，与在多肽序列的随机诱变之后发生的更表浅的诱变相反。例如，饱和诱变方法可用于突变“热点”，例如通过使用在这些位置含有NNt/g或NNt/c的寡聚物进行。在一个实例中，使用本发明提供的方法，在u-PA中鉴别了有助于增加底物特异性的如下热点：73、80、30和155(基于胰凝乳蛋白酶编号)。这些位置的突变可以例如通过使用对野生型或者模板蛋白酶序列在一或多个这些位点进行饱和诱变而实现，以产生用于随后筛选中的蛋白酶突变体集合。
2.嵌合形式的变体蛋白酶
本发明提供的变体蛋白酶可包括嵌合蛋白或融合蛋白。在一个实例中，蛋白酶融合蛋白包含蛋白酶蛋白的至少一个催化活性部分。在另一个实例中，蛋白酶融合蛋白包含蛋白酶的至少两或多个催化活性部分。在融合蛋白中，非蛋白酶多肽可以与蛋白酶多肽的N末端或C末端融合。在一个实施方案中，所述融合蛋白包含一个柔性肽接头或间隔序列，其分隔蛋白酶与非蛋白酶多肽。在另一个实施方案中，所述融合蛋白可包含一个标记或者可检测的多肽。标记和可检测的蛋白质的例子为本领域所已知，包括例如但不限于组氨酸标记、血凝素标记、myc标记或者荧光蛋白。在另一个实施方案中，所述融合蛋白是GST-蛋白酶融合蛋白，其中所述蛋白酶序列与GST序列(谷胱甘肽S-转移酶)的N末端融合。这种融合蛋白可以促进重组蛋白酶多肽的纯化。在另一个实施方案中，所述融合蛋白是Fc融合蛋白，其中所述蛋白酶序列与免疫球蛋白G的Fc结构域的N末端融合。这种融合蛋白可以具有较好的体内药物动力学性质。在另一个实施方案中，所述融合蛋白是在其N末端包含异源信号序列的蛋白酶。在某些宿主细胞中(例如哺乳动物宿主细胞)，蛋白酶的表达和/或分泌可以通过使用异源信号序列而增加。
蛋白酶嵌合蛋白或融合蛋白可以通过标准重组DNA技术产生。例如，利用常规技术将编码不同多肽序列的DNA片段符合读框地连接在一起，例如应用平末端或者交错末端进行连接、应用限制酶消化以提供合适的末端、适当地填充粘端、应用碱性磷酸酶处理以避免不希望的结合和酶连接。在另一个实施方案中，所述融合基因可以通过常规技术合成，包括利用自动DNA合成仪合成。或者，可以使用锚引物对基因片段进行PCR扩增，所述引物导致在两个连续基因之间产生互补突出端，其随后可退火和再扩增产生嵌合基因序列(见例如Ausubel et al.(eds.)CURRENT PROTOCOLS INMOLECULAR BIOLOGY，John Wiley & Sons，1992)。此外，可商购编码融合部分(例如GST多肽)的许多表达载体。可以将蛋白酶编码核酸克隆进这种表达载体中，由此所述融合部分与所述蛋白酶蛋白符合读框地连接。
3.组合文库及其它文库
筛选测定方法所用化合物的来源可以是例如文库的集合，包括但不限于组合文库。合成组合文库的方法和这种组合文库的特征为本领域所已知(见Combinatorial Libraries：Synthesis，Screening and Application Potential(Cortese Ed.)Walter de Gruyter，Inc.，1995；Tietze and Lieb，Curr.Opin.Chem.Biol.，2(3)：363-71(1998)；Lam，Anticancer Drug Des.，12(3)：145-67(1997)；Blaney and Martin，Curr.Opin.Chem.Biol.，1(1)：54-9(1997)；和Schultz andSchultz，Biotechnol.Prog.，12(6)：729-43(1996))。
使用分子生物学方法和/或同时使用化学合成方法已经开发了产生不同文库包括蛋白酶或酶文库、包括位置扫描合成组合文库(PSSCL)的方法和策略(见例如Georgiou，et al.(1997)Nat.Biotechnol.15：29-34；Kim et al.(2000)Appl Environ Microbiol.66：788 793；MacBeath，G.P.et al.(1998)Science279：1958-1961；Soumillion，P.L.et al.(1994)Appl.Biochem.Biotechnol.47：175-189，Wang，C.I.et al.(1996).Methods Enzymol.267：52-68，U.S.Patents 6,867,010，6,168,919，U.S.Patent Application No.2006-0024289)。获得的组合文库潜在地含有数百万种可以筛选的化合物，以鉴别呈现选择的活性的化合物。
在一个实例中，蛋白酶的集合或文库的化合物可以展示于遗传包装上，包括但不限于在可展示多肽部分的任何可复制的载体上展示，所述载体例如是噬菌体、病毒或者细菌。多个展示的多肽由遗传包装展示，由此使得所述多肽如蛋白酶或其催化活性部分可以结合和/或与靶多肽相互作用。举例的遗传包装包括但不限于噬菌体(见例如Clackson et 25a/.(1991)MakingAntibody Fragments Using Phage Display Libraries，Nature，352：624-628；Glaser et al.(1992)Antibody Engineering by Condon-Based Mutagenesis in aFilamentous Phage Vector System，J.Immunol.，149：3903 3913；Hoogenboom etal.(1991)Multi-Subunit Proteins on the Surface of Filamentous Phage：Methodologies for Displaying Antibody(Fate)Heavy and 30 Light Chains，Nucleic Acids Res.，19：4133-41370)、杆状病毒(见例如Boublik et a/.(1995)Eukaryotic Virus Display：Engineering the Major Surface Glycoproteins of theAutographa California Nuclear Polyhedrosis Virus(ACNPV)for thePresentation of Foreign Proteins on the Virus Surface，Bio/Technology，13：1079-1084)、细菌及用于展示蛋白质例如噬菌体展示的蛋白酶的其它合适载体。例如，感兴趣的噬菌体包括但不限于T4噬菌体、M13噬菌体和HI噬菌体。遗传包装任选在例如细菌宿主中扩增。任何这些遗传包装以及本领域技术人员已知的任何其它遗传包装均可用于本发明的方法中，以展示蛋白酶或其催化活性部分。
a.噬菌体展示文库
用于筛选的变体蛋白酶或其催化活性部分的文库可以在噬菌体表面上表达，所述噬菌体例如但不限于是M13、fd、f1、T7和λ噬菌体(见例如Santini(1998)J.Mol.Biol.282：125-135；Rosenberg et al.(1996)Innovations 6：1-6；Houshmand et al.(1999)Anal Biochem 268：363-370，Zanghi et al.(2005)Nuc.Acid Res.33(18)e160：1-8)。所述变体蛋白酶可以与噬菌体外壳蛋白以共价键、非共价键或者非肽键形式融合(见例如美国专利5,223,409，Crameri et al.(1993)Gene 137：69和WO 01/05950)。编码变体蛋白酶的核酸可以与编码外壳蛋白的核酸融合以产生蛋白酶-外壳蛋白融合蛋白，其中所述变体蛋白在所述噬菌体表面上表达。例如，编码变体蛋白酶的核酸可以与编码丝状噬菌体M13 Gene III(gIIIp；SEQ ID NO：512)的C末端结构域的核酸融合。在一些实例中，呈现在噬菌体上改良的展示的突变体蛋白酶用作模板以产生如本文所述突变体噬菌体展示文库。例如，如实施例8所述，在相应于野生型MT-SP1第122位(基于胰凝乳蛋白酶编号)的位置具有丝氨酸突变为半胱氨酸的突变体MT-SP1呈现改良的噬菌体展示。因此，这种突变体可以用作模板，从中产生文库多样性。
此外，所述融合蛋白可包含一个柔性肽接头或间隔序列、标记或可检测的多肽、蛋白酶位点或者另外的氨基酸修饰以改良所述融合蛋白的表达和/或效用。例如，蛋白酶位点的加入使得可以在选择程序之后有效地回收希望的噬菌体。标记和可检测的蛋白的例子为本领域所已知，其包括例如但不限于组氨酸标记、血凝素标记、myc标记或者荧光蛋白。在另一个实例中，编码蛋白酶-外壳蛋白融合体的核酸可以与前导序列融合以改良所述多肽的表达。前导序列的例子包括但不限于STII或OmpA。噬菌体展示在例如Ladner et al.，美国专利5,223,409；Rodi et al.(2002)Curr.Opin.Chem.Biol.6：92-96；Smith(1985)Science 228：1315-1317；WO 92/18619；WO91/17271；WO 92/20791；WO 92/15679；WO 93/01288；WO 92/01047；WO 92/09690；WO 90/02809；de Haard et al.(1999)J.Biol.Chem274：18218-30；Hoogenboom et al.(1998)Immunotechnology 4：1-20；Hoogenboom et al.(2000)Immunol Today 2：371-8；Fuchs et al.(1991)Bio/Technology 9：1370-1372；Hay et al.(1992)Hum Antibod Hybridomas3：81-85；Huse et al.(1989)Science 246：1275-1281；Griffiths et al.(1993)EMBO J 12：725-734；Hawkins et al.(1992)J Mol Biol 226：889-896；Clacksonet al.(1991)Nature 352：624-628；Gram et al.(1992)PNAS 89：3576-3580；Garrard et al.(1991)Bio/Technology 9：1373-1377；Rebar et al.(1996)MethodsEnzymol.267：129-49；Hoogenboom et al.(1991)Nuc AcidRes 19：4133-4137；和Barbas et al.(1991)PNAS 88：7978-7982中描述。
适于噬菌体展示的核酸例如噬菌体载体为本领域所已知(见例如Andris-Widhopf et al.(2000)J Immunol Methods，28：159-81；Armstrong et al.(1996)Academic Press，Kay et al.，Ed.pp.35-53；Corey et al.(1993)Gene128(1)：129-34；Cwirla et al.(1990)Proc Natl Acad Sci USA 87(16)：6378-82；Fowlkes et al.(1992)Biotechniques 13(3)：422-8；Hoogenboom et al.(1991)Nuc Acid Res 19(15)：4133-7；McCafferty et al.(1990)Nature 348(6301)：552-4；McConnell et al.(1994)Gene 151(1-2)：115-8；Scott and Smith(1990)Science249(4967)：386-90)。
编码蛋白酶-外壳蛋白融合蛋白、典型如上述产生的蛋白酶变体的核酸文库可以掺入噬菌体基因组中或者插入噬菌粒载体中。在噬菌粒系统中，编码被展示的蛋白质的核酸被提供在噬菌粒载体上，典型长度少于6000个核苷酸。所述噬菌粒载体包括噬菌体复制起点，由此当用辅助噬菌体例如M13K01或M13VCS感染携带质粒的细菌细胞时，该质粒掺入噬菌体颗粒中。然而，噬菌粒没有足够的噬菌体基因以在感染后产生稳定的噬菌体颗粒。这些噬菌体基因可以由辅助噬菌体提供。典型地，所述辅助噬菌体提供基因III外壳蛋白及为噬菌体复制和装配所需的其它噬菌体基因的完整拷贝。由于所述辅助噬菌体具有缺陷的复制起点，因此辅助噬菌体基因组未有效地掺入与具有野生型来源的质粒相关的噬菌体颗粒中。见例如美国专利5,821,047所述。所述噬菌粒基因组含有一个可选择的标记基因例如Amp.sup.R或者Kan.sup.R(分别对于氨苄青霉素或者卡那霉素抗性)以选择由所述文库成员感染的细胞。
在噬菌体展示的另一个实例中，可以使用携带编码当表达时足以产生感染性噬菌体颗粒的一组噬菌体基因的核酸、噬菌体包装信号和自主复制序列的载体。例如，载体可以是已经修饰为包括编码被展示的蛋白质的序列的噬菌体基因组。噬菌体展示载体可进一步包括可插入外源核酸序列的位点，如含有限制酶消化位点的多克隆位点。外源核酸序列例如编码噬菌体载体中展示蛋白质的序列可以与核糖体结合位点、信号序列(例如M13信号序列)和转录终止子序列连接。
载体可以通过标准克隆技术构建以含有编码包括蛋白酶和噬菌体外壳蛋白一部分的多肽且与可调节的启动子可操纵地连接的序列。在一些实例中，噬菌体展示载体包括两种核酸序列，它们编码噬菌体外壳蛋白的相同区域。例如，载体包括一种序列，其编码处于与编码展示蛋白可操纵连接的位置的区域，以及另一种序列，其编码处于编码外壳蛋白的功能噬菌体基因(例如野生型噬菌体基因)中的区域。野生型及融合外壳蛋白的表达通过降低每个噬菌体颗粒产生融合蛋白的量而有助于成熟噬菌体的产生。这种方法在噬菌体对融合蛋白的耐受性较低的情况下尤其有用。
噬菌体展示系统典型地利用丝状噬菌体如M13、fd和f1。在使用丝状噬菌体的一些实例中，展示蛋白与噬菌体外壳蛋白锚定结构域融合。融合蛋白可以与具有相同锚定结构域的另一种多肽例如外壳蛋白的野生型或内源拷贝共表达。可以用于蛋白质展示的噬菌体外壳蛋白包括(i)丝状噬菌体的次要外壳蛋白，如基因III蛋白(gIIIp)，和(ii)丝状噬菌体的主要外壳蛋白，如基因VIII蛋白(gVIIIp)。也可以使用与其它噬菌体外壳蛋白如基因VI蛋白、基因VII蛋白或基因IX蛋白的融合体(见例如WO 00/71694)。
也可以使用这些蛋白质的部分(例如结构域或片段)。有用的部分包括稳定掺入噬菌体颗粒中的结构域，例如如此融合蛋白在整个选择程序中保持在颗粒中。在一个实例中，使用gIIIp的锚定结构域(见例如美国专利5,658,727及以下实施例)。在另一个实例中，使用gVIIIp(见例如美国专利5,223,409)，其可以是与展示蛋白融合的成熟的全长gVIIIp。丝状噬菌体展示系统典型地使用蛋白质融合体以将异源氨基酸序列附着于噬菌体外壳蛋白或锚定结构域。例如，噬菌体可以包括编码信号序列、异源氨基酸序列和锚定结构域例如gIIIp锚定结构域的基因。
表达的融合蛋白的价可以通过选择噬菌体外壳蛋白而控制。例如，gIIIp蛋白典型地以每个病毒粒3-5个拷贝掺入噬菌体外壳中。gIIIp与变体蛋白酶的融合体由此产生低价。相比之下，gVIII蛋白典型地以每个病毒粒2700个拷贝掺入噬菌体外壳中(Marvin(1998)Curr.Opin.Struct.Biol.8：150-158)。由于gVIIIp的高价，噬菌体通常对大于10个残基的肽耐受不良。可以使用噬菌粒系统通过提供外壳蛋白的野生型拷贝以降低融合蛋白的价而增加噬菌体对更大的肽的耐受性。此外，可以使用gVIIIp的突变体，它们被优化表达更大的肽。在一个这种实例中，在gVIIIp的诱变筛选中获得了突变体gVIIp，其具有改良的表面展示性质(Sidhu et al.(2000)J.Mol.Biol.296：487-495)。
也可以使用可调节的启动子来控制展示蛋白的价。可以使用调节表达来产生具有低价展示蛋白的噬菌体。许多可调节的(例如可诱导的和/或可阻遏的)启动子序列是已知的。这类序列包括可调节的启动子，其活性可以通过用户的干预例如通过操纵环境参数例如温度或通过加入刺激分子或去除阻遏分子而被改变或调节。例如，可以加入外源化合物调节一些启动子的转录。可调节的启动子可以含有一或多种转录激活物或阻遏物蛋白的结合位点。包括转录因子结合位点的合成启动子可以被构建并也可以被用作可调节的启动子。举例性的可调节启动子包括应答环境参数例如热变化、激素、金属、代谢物、抗生素或化学物质的启动子。适于在大肠杆菌中使用的可调节启动子包括含有来自lac，tac，trp，trc和tet操纵子序列或操纵子的转录因子结合位点的启动子，碱性磷酸酶启动子(pho)，阿拉伯糖启动子如araBAD启动子，鼠李糖启动子，启动子本身或其功能片段(见例如Elvinet al.(1990)Gene 37：123-126；Tabor and Richardson，(1998)Proc.Natl.Acad.Sci.U.S.A.1074-1078；Chang et al.(1986)Gene 44：121-125；Lutz and Bujard，(1997)Nucl.Acids.Res.25：1203-1210；D.V Goeddel et al.(1979)Proc.Nat.Acad.Sci.U.S.A.，76：106-110；J.D.Windass et al.(1982)Nucl.Acids.Res.，10：6639-57；R.Crowl et al.(1985)Gene，38：31-38；Brosius(1984)Gene 27：161-172；Amanna and Brosius，(1985)Gene 40：183-190；Guzman et al.(1992)J.Bacteriol.，174：7716-7728；Haldimann et al.(1998)J.Bacteriol.，180：1277-1286)。
例如，lac启动子可以被乳糖或结构相关分子如异丙基-β-D-硫代半乳糖苷(IPTG)诱导并由葡萄糖阻遏。一些可诱导的启动子通过去阻遏过程例如阻遏物分子的失活而诱导。
可调节的启动子序列也可以被间接调节。可以被工程化而间接调节的启动子的例子包括：噬菌体λPR，PL，噬菌体T7，SP6和T5启动子。例如，调节序列通过其表达被例如环境参数调节的因子而阻遏或激活。这种启动子的一个例子是T7启动子。T7RNA聚合酶的表达可以被环境应答启动子如lac启动子调节。例如，细胞可以包括异源核酸，该异源核酸包括编码T7 RNA聚合酶的序列和被环境参数调节的调节序列(例如lac启动子)。T7 RNA聚合酶的活性也可以被RNA聚合酶的天然抑制剂如T7溶菌酶的存在而调节。
在另一种构型中，λPL可以被工程化而由环境参数调节。例如，细胞可以包括核酸序列，该核酸序列编码λ阻遏物的温度敏感变体。升高细胞至非允许温度使PL启动子从阻遏中释放。
启动子或转录调节序列的调节性质可以通过将所述启动子或序列可操纵地连接至编码报道蛋白(或任何可检测蛋白)的序列而容易地被测试。这种启动子-报道融合序列典型地在质粒或载体中被导入细菌细胞中，报道蛋白的丰度在各种环境条件下评估。有用的启动子或序列是在某些条件下被选择性激活或阻遏的启动子或序列。
在一些实施方案中，使用非可调节的启动子。例如，可以选择这样的启动子，其在相关条件下产生合适量的转录。非可调节的启动子的例子是gIII启动子。
b.细胞表面展示文库
用于筛选的变体蛋白酶文库可以在细胞例如原核或真核细胞的表面上表达。用于细胞表面表达的举例性的细胞包括但不限于细菌、酵母、昆虫细胞、禽类细胞、植物细胞和哺乳动物细胞(Chen and Georgiou(2002)Biotechnol Bioeng 79：496-503)。在一个实例中，用于表达的细菌细胞是大肠杆菌。
变体蛋白酶可以作为与在细胞表面上表达的蛋白质如膜蛋白或细胞表面相关蛋白的融合蛋白而表达。例如，变体蛋白酶可以在大肠杆菌中作为与大肠杆菌外膜蛋白(例如OmpA)的融合蛋白而表达，一种遗传工程化的主要大肠杆菌脂蛋白(Lpp)和外膜蛋白OmpA或细胞表面相关蛋白(例如菌毛或鞭毛亚基)的杂种分子。通常，当细菌外膜蛋白被用于异源肽或蛋白质的展示时，其通过遗传插入载体蛋白的允许位点而实现。异源肽或蛋白质的表达依赖于插入的蛋白结构域的结构性质，因为所述肽或蛋白质当被插入到允许位点时与融合在蛋白质的N末端或C末端相比更受限。可以对融合蛋白进行修饰以改善融合蛋白的表达，如插入柔性肽接头或间隔序列或修饰细菌蛋白(例如通过氨基酸序列中的突变、插入或缺失)。酶如粪肥纤维单胞菌(Cellulomonas fimi)的β-内酰胺酶和Cex外切葡聚糖酶已被成功在大肠杆菌表面上表达为Lpp-OmpA融合蛋白(Francisco J.A.andGeorgiou G.Ann N Y Acad Sci.745：372-382(1994)和Georgiou G.et al.Protein Eng.9：239-247(1996))。15-514个氨基酸的其它肽已经被展示在OmpA表面上的第二、第三和第四个外环中(Samuelson et a1.J.Biotechnol.96：129-154(2002))。因此外膜蛋白可以在细菌外表面上携带和展示异源基因产物。
在另一个实例中，变体蛋白酶可以与蛋白质的自转运蛋白结构域融合，所述蛋白质如淋球菌(N.gonorrhoeae)IgA1蛋白酶、粘质沙雷氏菌(Serratiamarcescens)丝氨酸蛋白酶、福氏志贺菌(Shigella flexneri)VirG蛋白和大肠杆菌粘附素AIDA-I(Klauser et al.EMBO J.1991-1999(1990)；Shikata S，et al.J Biochem.114：723-731(1993)；Suzuki T et al.J Biol Chem.270：30874-30880(1995)；和Maurer J et al.J Bacteriol.179：794-804(1997))。其它自转运蛋白包括革兰氏阴性菌种(例如大肠杆菌，沙门氏菌鼠伤寒血清变种和福氏志贺菌)中存在的那些。酶如β-内酰胺酶已经使用这一系统被成功在大肠杆菌表面上表达(Lattemann CT et al.J Bacteriol.182(13)：3726-3733(2000))。
细菌可以被重组工程化以表达融合蛋白如膜融合蛋白。编码变体蛋白酶的核酸可以与编码细胞表面蛋白例如但不限于细菌OmpA蛋白的核酸融合。编码变体蛋白酶的核酸可以被插入膜蛋白的允许位点，如膜蛋白的细胞外环。此外，编码融合蛋白的核酸可以与编码标记或可检测蛋白的核酸融合。这种标记和可检测蛋白是本领域已知的并包括例如但不限于组氨酸标记、血凝素标记、myc标记或荧光蛋白。编码融合蛋白的核酸可以可操纵地与启动子连接以在细菌中表达。例如，可以将核酸插入到携带用于表达融合蛋白的启动子和任选地用于选择的另外的基因如抗生素抗性基因的载体或质粒中。细菌可以用这种质粒转化，如通过电穿孔或化学转化而转化。这种技术是本领域技术人员已知的。
外膜或周质空间中的蛋白质通常在细胞质中作为未成熟蛋白合成，所述未成熟蛋白在信号序列处被裂解而产生成熟蛋白被运输到细胞质外。用于大肠杆菌的重组蛋白的分泌产生的举例性信号序列是已知的。当感兴趣的基因被正确地与信号序列融合时，没有Met延伸的N末端氨基酸序列可以在用信号肽酶裂解后获得。因此，成熟蛋白可以无需改变感兴趣蛋白质的氨基酸序列而产生(Choi and Lee.Appl.Microbiol.Biotechnol.64：625-635(2004))。
本领域已知其它的细胞表面展示系统，包括但不限于基于冰核蛋白(Inp)的细菌表面展示系统(Lebeault J M(1998)Nat Biotechnol.16：57680)，酵母展示(例如与酵母Aga2p细胞壁蛋白的融合；参见美国专利6,423,538)，昆虫细胞展示(例如杆状病毒展示，参见Ernst et al.(1998)Nucleic AcidsResearch，Vol 26，Issue 7 1718-1723)，哺乳动物细胞展示，以及其它真核展示系统(见例如5,789,208和WO 03/029456).
c.其它展示文库
还可以使用其它展示形式以筛选变体蛋白酶的文库，例如根据本文所述设计变异的文库。举例性的其它展示形式包括核酸-蛋白质融合体，核酶展示(见例如Hanes and Pluckthun(1997)Proc.Natl.Acad.Sci.U.S.A.13：4937-4942)，珠展示(Lam，K.S.et al.Nature(1991)354，82-84；，K.S.etal.(1991)Nature，354，82-84；Houghten，R.A.et al.(1991)Nature，354，84-86；Furka，A.et al.(1991)Int.J.Peptide Protein Res.37，487-493；Lam，K.S.，et al.(1997)Chem.Rev.，97，411-448；美国公布专利申请2004-0235054)和蛋白质阵列(见例如Cahill(2001)J.Immunol.Meth.250：81-91，WO01/40803，WO 99/51773，和US2002-0192673-A1)。
在特异性的其它例子中，可以有利地将蛋白酶、变体蛋白酶、或催化活性部分或噬菌体文库或表达变体蛋白酶的细胞附着于固体支持物。例如，在一些实例中，表达变体蛋白酶的细胞可以天然吸附到珠上，由此一群珠中每个珠含有一个单细胞(Freeman et al.Biotechnol.Bioeng.(2004)86：196-200)。在固定化到玻璃支持物上之后，可以生长微菌落并用生色或荧光底物筛选。在另一个实例中，变体蛋白酶或噬菌体文库或表达变体蛋白酶的细胞可以列阵在滴定板中并固定化。
F.接触、分离和鉴别所选择的蛋白酶的方法
在已经选择和制备展示蛋白酶或其催化活性部分的多个集合或文库之后，所述文库用于使靶蛋白酶捕获多肽与蛋白酶成分接触。将靶底物与被展示的蛋白酶文库接触以选择具有改变的底物特异性的蛋白酶，所述靶底物包括例如蛋白酶捕获多肽如在其RSL环中突变而具有希望的裂解序列的丝氨酸蛋白酶抑制蛋白。蛋白酶和蛋白酶捕获多肽可以在悬浮液、溶液中接触，或者通过固体支持物接触。使所述成分以足够的时间、温度或浓度接触，以发生相互作用和随后的裂解反应及选择的蛋白酶与蛋白酶捕获多肽的稳定的中间体复合物形成。反应保持的严格性可以通过改变一或多个参数而调节，所述参数包括反应温度、蛋白酶捕获多肽抑制剂的浓度、竞争剂(如果有的话)的浓度、混合物中蛋白酶集合的浓度和保温时间长度。
与蛋白酶捕获多肽形成共价复合物的选择的蛋白酶被捕获和分离。为了便于捕获，用于筛选的蛋白酶捕获多肽可提供在溶液、悬浮液中，或者附着于固体支持物上以适用于测定方法。例如，蛋白酶捕获多肽可以附着于固体支持物上，例如一或多个如下固体支持物：珠或颗粒、微球体、试管或平板的表面、滤膜及本领域已知的其它固体支持物。固体支持系统的例子包括但不限于构建的平面，例如玻璃、硅、金属、尼龙、纤维素、塑料或者合成物，包括多孔平板或膜；或者可以是珠形式，例如硅凝胶、受控的有孔玻璃、磁珠(Dynabead)或者纤维素珠。对这种方法可以加以修改以适用于悬浮液或者柱形式。靶蛋白酶捕获多肽可以直接或间接附着于固体支持物，如聚丙烯酰胺珠。可以使用共价或非共价方法进行附着。靶化合物的共价附着方法包括化学交联方法。反应试剂可以在靶分子与支持物上的官能团之间产生共价键。可以化学反应的官能团的例子是氨基、硫醇和羧基基团。N-乙基马来酰亚胺、碘乙酰胺、N-羟基琥珀酰亚胺和戊二醛是与官能团反应的试剂的例子。在其它实例中，靶底物可以间接附着于固体支持物上，通过例如但不限于免疫亲和性或者配体-受体相互作用(例如生物素-链霉抗生物素或者谷胱甘肽S-转移酶-谷胱甘肽)等方法附着。例如，蛋白酶捕获多肽可以被包被于ELISA平板或者其它相似的可寻址阵列上。在一个实例中，平板的孔可以用亲和性捕获剂包被，所述亲和性捕获剂结合且捕获蛋白酶捕获多肽。实施例9例证了将生物素化的抗-His抗体包被于含有链霉抗生物素的平板上以便于含有His标记的蛋白酶捕获多肽的捕获。
蛋白酶捕获多肽与固体支持物的附着可以在其与变体蛋白酶或者噬菌体文库或者表达变体蛋白酶的细胞接触之前、期间或者之后进行。例如，靶底物可以预先附着于固体支持物上，例如附着于层析柱，之后与变体蛋白酶保温。在其它实例中，在靶底物与变体蛋白酶结合之后再附着于固体支持物。
在这样的实例中，可以洗涤含有复合的底物-蛋白酶对的固体支持物以除去任何未结合的蛋白酶。所述复合物可以通过本领域技术人员已知的任何方法从固体支持物中回收，例如通过用稀酸处理，随后进行中和(Fu et al.(1997)J Biol.Chem.272：25678-25684)或用三乙胺处理(Chiswell et al.(1992)Trends Biotechnol.10：80-84)。这个步骤可以优化为保证从固体底物中可再生地和定量回收所述展示源。例如，所述展示源与附着于固体支持物的靶底物的结合可以使用本领域技术人员熟知的方法单独监测，例如使用针对噬菌体如M13噬菌体的抗体(例如New England Biolabs，MA)和标准ELISA(见例如Ausubel et al.(1987)Current Protocols in Molecular Biology，John Wiley& Sons，New York)监测。
另一种捕获和分离底物-蛋白酶复合物的方法是从溶液中捕获和分离。典型地，在这种方法中，将蛋白酶捕获多肽或其变体与蛋白酶集合接触，例如在小体积(即20、30、40、50、60、70、80、90、100、200、300、400、500或更多微升)的合适结合缓冲液中接触，其中每个蛋白酶捕获多肽均与预定的标记(marker)、标记(tag)或者其它可检测的部分结合以进行鉴别和分离。所述可检测的部分可以是便于底物-蛋白酶复合物的检测和分离的任何部分。例如，所述部分可以是表位标记，为此存在特异于该标记的抗体(即myc-标记，His-标记或者其它标记)。所述抗体可以结合固体支持物如珠以便于捕获稳定复合物。可以使用其它相似策略，包括例如用生物素标记靶底物，使用与固体支持物如磁珠或微滴定平板附着的或者用聚组氨酸(例如His6-标记)标记链霉抗生物素进行捕获以及使用金属螯合剂例如但不限于硫酸镍(NiSO4)、氯化钴(CoCl2)、硫酸铜(CuSO4)或者氯化锌(ZnCl2)进行捕获。捕获剂可以与大的珠例如琼脂糖珠结合，从而可以简单地通过离心即可分离结合的珠。或者，捕获剂可以与较小的珠例如磁珠(即Miltenyi Biotec)偶联，其可以通过磁性柱简单分离。另外，所述部分可以是荧光部分。例如，在一些展示系统中，例如在细胞表面展示系统中，荧光标记可以便于通过荧光激活细胞分选法(FACS见例如Levin et al.(2006)Molecular BioSystems，2：49-57)分离选择的复合物。
在一些情况中，将一或多个独特的蛋白酶捕获多肽与蛋白酶集合接触，其中每个蛋白酶捕获多肽均与不同的检测部分结合，以便单独分离一个或一个以上的蛋白酶捕获多肽-蛋白酶复合物。在一个反应中包含1、2、3、4、5、6、7、8、9、10或更多个独特蛋白酶捕获多肽(每个多肽均具有不同的希望的RSL裂解序列)的能力使得可以同时检测和分离数十、数百或数千个共价复合物。
作为与蛋白酶捕获多肽的共价复合物而被捕获的选择的蛋白酶可以与蛋白酶集合中未复合的蛋白酶分开。然后可以扩增选择的蛋白酶以便于鉴别选择的蛋白酶。在除去未与蛋白酶捕获多肽复合的任何未复合的蛋白酶之后，展示所述蛋白酶的材料的来源(即噬菌体、细胞、珠等)被扩增并在适当宿主细胞中表达。例如，在蛋白酶被展示在噬菌体上的情况中，通常蛋白酶捕获多肽复合物中的蛋白酶-噬菌体与宿主细胞保温以使得噬菌体吸附，随后加入小体积的营养肉汤并搅拌所述培养物以便于噬菌体探针DNA在扩增宿主中复制。在一些实例中，这是在存在辅助噬菌体的条件下进行的，以保证宿主细胞被所述噬菌体感染。在保温之后，为培养基补加抗生素和/或诱导剂。噬菌体蛋白酶基因组也可以含有编码抗生素抗性的基因，以使得保留所述噬菌体蛋白酶DNA的那些细菌细胞可以选择性生长。典型地，为了扩增作为含有选择的蛋白酶的噬菌体上清来源的噬菌体，需要通过使用辅助噬菌体以拯救所述噬菌体。在一些实例中，可以不进行拯救步骤测定选择的蛋白酶的存在。例如，在含有选择的或鉴别的蛋白酶的被捕获的复合物与宿主细胞例如细菌保温、并在存在选择性物质的条件下生长之后，可以将细胞周质或者细胞培养基直接取样，作为选择的蛋白酶的来源以例如测量蛋白酶活性。这个程序在实施例17中描述。
此外，展示源在例如细菌宿主中的扩增可以通过多种方式优化。例如，加入到测定材料如微孔中的细菌的量可以显著超过从结合步骤回收的噬菌体源的量，从而保证噬菌体基因组的定量转导。转导效率任选可以在选择噬菌体时进行测量。所述扩增步骤扩增了展示源的基因组如噬菌体基因组，使得相关的特征多肽过表达和得以鉴别例如通过DNA测序鉴别。
可以使用淘选方法，由此快速选择与靶蛋白相互作用的蛋白酶或其催化活性部分，所述靶蛋白例如是蛋白酶捕获多肽或其RSL变体。淘选例如通过如下方式进行：将噬菌体展示多肽文库如噬菌体展示蛋白酶与表面结合的或者可溶的靶蛋白保温，洗涤去未结合的噬菌体，洗脱特异性和共价结合的噬菌体。然后例如通过感染宿主而扩增洗脱的噬菌体，通过另外的淘选与扩增循环以继续富集对于靶多肽具有最高亲和性的那些噬菌体库(pool)。在几次循环之后，通过例如DNA测序鉴别单独的克隆，其活性可以通过例如下文G章节所述的任何方法测量。
一旦鉴别了选择的蛋白酶，可以将其从展示源中纯化并检测活性。通常，这种方法包括一般的生物化学方法和重组DNA技术，且为本领域技术人员所熟知。在一种方法中，聚乙二醇(PEG)沉淀法可以用于除去纯化的选择的噬菌体上清中潜在污染蛋白酶活性。在这种实例中，在辅助噬菌体的存在下进行噬菌体拯救之后，含有选择的蛋白酶的噬菌体上清可以在PEG的存在下被沉淀。本领域技术人员能确定为特定沉淀反应所需的PEG的百分比。通常，为了沉淀蛋白酶上清，使用20％PEG。
在一些实例中，可以如本文所述测定上清的蛋白酶活性，所述上清来自拯救的噬菌体上清或者来自细菌细胞周质或者细胞培养基(不进行噬菌体拯救)。或者或另外，选择的蛋白酶可以纯化自所述上清或其它来源。例如，编码选择的蛋白酶结构域的DNA可以分离自展示源，使得可以纯化选择的蛋白质。例如，在如上述用选择的噬菌体感染大肠杆菌宿主细胞之后，可以挑取单独的克隆并使其生长以使用本领域技术人员已知的任何方法进行质粒纯化，如果需要则可以大量制备，例如使用Midi质粒纯化试剂盒(Qiagen)进行。纯化的质粒可用于DNA测序以鉴别变体蛋白酶的序列；或者可以用于转染进任何细胞中以进行表达，例如但不限于转染进哺乳动物表达系统中。如果需要，可以进行一或两步PCR以扩增选择的序列，其可以被亚克隆进选择的表达载体中。可以设计PCR引物以便于亚克隆，例如包括加入限制酶位点。实施例4举例说明了两步PCR方法以完成全长u-PA基因的扩增与纯化，其中选择的蛋白酶噬菌体仅含有u-PA基因的蛋白酶结构域。如下文详细描述，在转染进合适的细胞中表达之后，含有蛋白酶多肽或其催化活性部分的条件培养基可以在活性测定中检测或者可用于进一步纯化。另外，如果需要，可以对蛋白酶进行加工以产生活性蛋白酶，例如通过单链形式裂解为双链形式而产生活性蛋白酶。这种操纵为本领域技术人员所已知。例如，单链u-PA可以通过纤溶酶的裂解而被激活，如本文所述。
1.重复筛选
在本发明提供的方法中，重复筛选用于优化蛋白酶的修饰。因此，在重复筛选方法中，通过在各种参数下进行多次淘选反应而可以使蛋白酶进化，例如通过使用不同的蛋白酶捕获多肽或竞争剂。在这种重复筛选方法中，蛋白酶集合在连续的筛选循环中可以保持恒定。或者，可以产生仅含有在之前的循环中鉴别的选择的蛋白酶的新的蛋白酶集合和/或产生新的突变体蛋白酶集合，其与在第一次循环中鉴别的模板蛋白酶相比已经被进一步突变。
在一个实例中，对蛋白酶文库进行的第一次筛选循环可以鉴别变体蛋白酶，所述变体蛋白酶含有改变蛋白酶特异性的一或多个突变。然后可以进行第二轮文库合成，其中所述一或多个突变的氨基酸位置保持恒定，对蛋白质其余部分或者希望的区域或残基进行聚焦或随机诱变。在另一轮筛选之后，可以对选择的蛋白酶进行另外的文库合成与筛选循环。例如，可以进行2、3、4、5或更多轮文库合成与筛选。在一些实例中，变体蛋白酶对于改变的底物的特异性通过每次选择循环而得以进一步优化。
在重复筛选的另一种方法中，对蛋白酶集合的第一轮筛选可以针对中间体蛋白酶捕获多肽进行以鉴别含有改变蛋白酶对于中间体底物的特异性的一或多个突变的变体蛋白酶。可以分离选择的蛋白酶复合物，使其生长，并且在合适的宿主细胞中扩增，且在第二轮筛选中用作蛋白酶集合，所述第二轮筛选针对含有靶多肽的完整裂解序列的蛋白酶捕获多肽进行。例如，这种方法可用于选择对于VEGFR裂解序列具有底物特异性的蛋白酶，其中一或多轮淘选循环针对RRARM中间体裂解序列进行，随后的淘选循环针对含有VEGFR2裂解序列RRVR的蛋白酶捕获多肽进行。
在重复筛选的另一个实例中，含有对于两或多个不同多肽具有不同的底物识别或裂解序列的两或多个蛋白酶捕获多肽用于交替的淘选循环方法中。这种方法可用于选择优化为对于两种不同底物具有选择性的蛋白酶。选择的变体对于两或多个底物识别序列典型具有窄的特异性，但是具有高活性。在这种方法中，针对第一种蛋白酶捕获多肽(其已经被修饰以选择具有第一种预定底物特异性的蛋白酶)进行的第一轮蛋白酶集合筛选可以鉴别含有改变蛋白酶特异性的一或多个突变的变体蛋白酶。可以分离选择的蛋白酶，使其生长，并且在合适宿主细胞中扩增，且在第二轮筛选中用作蛋白酶集合，所述第二轮筛选针对已经被修饰以选择具有第二种预定底物特异性的蛋白酶的第二种蛋白酶捕获多肽进行。本方法中使用的第一种和第二种蛋白酶捕获多肽可以相同或不同，但每种在其反应位点中均不同修饰以模拟不同靶底物的底物识别位点(即裂解序列)。在一些实例中，选择的严格性可以在存在竞争剂的条件下增强，所述竞争剂例如是如本文所述的窄谱或广谱竞争剂。
2.举例性的选择的蛋白酶
本发明提供了在本发明的方法中鉴别的变体u-PA和MT-SP1多肽，其具有改变的和/或改良的底物特异性。鉴别的这种变体u-PA和MT-SP1多肽对于选择的或者希望的靶蛋白的裂解序列具有增加的特异性。举例的这种靶蛋白包括但不限于VEGFR或补体蛋白例如补体蛋白C2中的裂解序列。任何修饰的丝氨酸蛋白酶抑制蛋白均可以用于本发明的选择方法中以鉴别变体蛋白酶。这种修饰的丝氨酸蛋白酶抑制蛋白的例子是PAI-1或AT3，在其RSL中被修饰为含有靶蛋白例如VEGFR或C2的裂解序列，如本文如上所述。获得的选择的修饰的蛋白酶与模板或起始蛋白酶相比呈现出对于靶蛋白中裂解序列的底物特异性被改变，典型地是被改良，所述模板或起始蛋白酶不含有选择的修饰。如下文所述，当与野生型或模板蛋白酶对于针对非靶底物选择的靶底物的特异性相比时，所述特异性典型地增加，通常增加至少2倍、至少4倍、至少5倍、至少6倍、至少7倍、8、9、10、20、30、40、50、60、70、80、90、100、200、300、400、500、600、700、800、900、1000倍或更多倍。
a.变体u-PA多肽
例如，本发明提供的变体u-PA多肽是根据对于突变体丝氨酸蛋白酶抑制蛋白多肽具有增加的反应性而加以选择的，所述突变体丝氨酸蛋白酶抑制蛋白多肽是在其RSL序列中通过用希望或选择的靶蛋白的氨基酸置换天然P4-P1’反应位点氨基酸而加以修饰。在一个实例中，变体u-PA多肽针对修饰的PAI-1多肽的选择而鉴别。本发明提供的u-PA选择方法中使用的修饰的PAI-1多肽分子的例子包括例如这样修饰的PAI-1，其天然P4-P1’残基VSARM(SEQ ID NO：378)用中间体VEGFR-2裂解序列RRARM(SEQ IDNO：379)的氨基酸残基修饰，其中P4-P1位置中希望的裂解序列是VEGFR-2裂解序列RRVR(SEQ ID NO：489)，或者用最佳的t-PA裂解序列PFGRS(SEQ ID NO：389)的氨基酸残基修饰。
使用本发明提供的方法鉴别了如下位置有助于u-PA多肽的底物特异性：21、24、30、38、39、61(A)、72、73、75、80、82、84、89、92、132、133、137、138、155、156、158、159、160、187和217(基于胰凝乳蛋白酶编号)。可以在相应于如下任一位置的任一或多个位置进行氨基酸置换：u-PA多肽如SEQ ID NO：433所示u-PA多肽或其催化活性部分的F21、I24、F30、V38、T39、Y61(A)、R72、L73、S75、E80、K82、E84、I89、K92、F132、G133、E137、I138、L155、K156、T158、V159、V160、K187和R217(基于胰凝乳蛋白酶编号)。本发明提供的呈现增加的底物特异性的修饰的u-PA多肽可以含有相应于如下任一或多个修饰的一或多个氨基酸修饰：u-PA多肽例如SEQ ID NO：433所示u-PA多肽或其催化活性部分的F21V、I24L、F30I、F30V、F30L、F30T、F30G、F30M、V38D、T39A、Y61(A)H、R72G、L73A、L73P、S75P、E80G、K82E、E84K、I89V、K92E、F132L、G133D、E137G、I138T、L155P、L155V、L155M、K156Y、T158A、V159A、V160A、K187E，和R217C(基于胰凝乳蛋白酶编号)。
在一个实例中，本发明提供的对于VEGFR-2裂解序列具有增加的底物特异性的修饰的u-PA多肽含有相应于如下任一或多个修饰的一或多个氨基酸修饰：V38D、F30I、F30T、F30L、F30V、F30G、F30M、R72G、L73A、L73P、S75P、I89V、F132L、G133D、E137G、I138T、L155P、L155V、L155M、V160A和R217C(基于胰凝乳蛋白酶编号)。这种多肽的例子是含有相应于如下任何修饰的一或多个氨基酸修饰的那些u-PA多肽：u-PA多肽例如具有SEQ ID NO：433所示氨基酸序列的u-PA多肽或其催化活性片段中的F30I；L73A/I89V；L73P；R217C；L155P；S75P/I89V/I138T；E137G；R72G/L155P；G133D；V160A；V38D；F132L/V160A；L73A/I89V/F30T；L73A/I89V/F30L；L73A/I89V/F30V；L73A/I89V/F30G；L73A/I89V/L155V；L73A/I89V/F30M；L73A/I89V/L155M；L73A/I89V/F30L/L155M；和L73A/I89V/F30G/L155M。这种序列的例子是SEQ ID NO：434-459所示任何序列，或者其含有所述突变且具有催化活性的连续氨基酸的片段。特别地，本发明提供了具有如下氨基酸修饰的修饰的u-PA多肽：L73A/I89V；L155P；R72G/L155P；F132L/V160A；L73A/I89V/F30T；L73A/I89V/L155V；L73A/I89V/L155M；和L73A/I89V/F30L/L155M(基于胰凝乳蛋白酶编号)。
在另一个实例中，本发明提供的对于由t-PA识别的裂解序列的特异性增加的修饰的u-PA多肽含有相应于如下任一或多个修饰的一或多个氨基酸修饰：F21V、I24L、F30V、F30L、T39A、Y61(A)H、E80G、K82E、E84K、I89V、K92E、K156T、T158A、V159A和K187E(基于胰凝乳蛋白酶编号)。这种多肽的例子是含有相应于如下任何修饰的一或多个氨基酸修饰的那些u-PA多肽：u-PA多肽例如具有SEQ ID NO：433所示氨基酸序列的u-PA多肽或其催化活性片段中的F21V；I24L；F30V；F30L；F30V/Y61(A)H；F30V/K82E；F30V/K156T；F30V/K82E/V159A；F30V/K82E/T39A/V159A；F30V/K82E/T158A/V159A；F30V/Y61(A)H/K92E；
F30V/K82E/V159A/E80G/I89V/K187E；和
F30V/K82EN159A/E80G/E84K/I89V/K187E。这种序列的例子是SEQ ID NO：460-472所示任何序列或者其含有所述突变且具有催化活性的连续氨基酸片段。
本发明还提供了胰凝乳蛋白酶家族的变体蛋白酶，其与本发明提供的变体u-PA多肽相比具有相应突变(基于胰凝乳蛋白酶编号)。例如，基于胰凝乳蛋白酶编号，u-PA中F30位置的修饰相应于t-PA中Q30、胰蛋白酶中Q30和胰凝乳蛋白酶中Q30位置中的修饰(Bode et al.(1997)Current Opinionin Structural Biology，7：865-872)。本领域技术人员可以确定任何其它胰凝乳蛋白酶家族成员中的相应突变，包括但不限于表7中列出的任何蛋白酶修饰且具有SEQ ID NO：40、42、44、46、48、50、52、54、56、58、60、62、64、66、68、70、72、74、76、78、80、82、84、86、88、90、92、94、96、98、99、101、103、105、107、109、111、113、115、117、119、121、123、125、127、129、131、133、135、137、139、141、143、145、147、149、151、153、155、157、159、161、163、165、167、169、171、173、175、177、179、181、183、185、187、189、193、195、197、199、201、203、205、207、209、211、213、215、217、219、221、223、225、227、229、231、233、235、237、239、241、242、243、245、247、249、251、253、255、257、259、261、262、264、266、268、270、272任一所示氨基酸序列或其催化活性片段。
b.变体MT-SP1多肽
在另一个实例中，本发明提供的变体MT-SP1多肽是根据对于突变体丝氨酸蛋白酶抑制蛋白多肽具有增加的反应性而选择的，所述突变体丝氨酸蛋白酶抑制蛋白多肽是在其RSL序列中通过用希望的或选择的靶蛋白的氨基酸置换天然P4-P2’反应位点氨基酸而修饰。在一个实例中，变体MT-SP1多肽是针对修饰的AT3多肽的选择而鉴别的。本发明提供的MT-SP1选择方法中使用的修饰的AT3多肽的例子包括例如这样修饰的AT3，即其天然P4-P2’残基IAGRSL(SEQ ID NO：478)用补体蛋白C2裂解序列SLGRKI(SEQ ID NO：479)的氨基酸残基修饰。
使用本发明提供的方法鉴别了如下位置有助于MT-SP1多肽的底物特异性：23、41、52、60(g)、65、71、93、95、97、98、99、126、129、131、136、143、144、154、164、166、171、173、175、184(a)、192、201、209、217、221(a)、230、234和244(基于胰凝乳蛋白酶编号)。氨基酸置换可以是在相应于如下任何位置的任一或多个位置进行：MT-SP1多肽例如SEQ IDNO：253或515所示全长MT-SP1多肽或者SEQ ID NO：505或507所示其催化活性部分中的D23、I41、L52、Y60(g)、T65、H71、F93、N95、F97、I98、F99、A126、V129、P131、I136、H143、T144、I154、N164、T166、L171、P173、Q175、F184(a)、Q192、S201、Q209、D217、Q221(a)、R230、F234和V244(基于胰凝乳蛋白酶编号)。本发明提供的呈现增加的底物特异性的修饰的MT-SP1多肽可含有相应于如下任一或多个修饰的一或多个氨基酸修饰：MT-SP1多肽例如SEQ ID NO：253或515所示全长MT-SP1多肽或者SEQ ID NO：505或507所示其催化活性部分中的D23E、I41F、I41T、L52M、Y60(g)s、T56K、H71R、F93L、N95K、F97Y、F97L、T98P、F99L、A126T、V129D、P131S、I136T、I136V、H143R、T144I、I154V、N164D、T166A、L171F、P173S、Q175R、F184(a)L、Q192H、S201I、Q209L、D217V、Q221(a)L、R230W、F234L和V244G(基于胰凝乳蛋白酶编号)。特别地，修饰的MT-SP1多肽含有相应于I41F、F97Y、L171F、Q175R、D217V和V244G的任一或多个修饰的一或多个氨基酸修饰，例如I41F、F97Y、L171F和V244G的任一或多个修饰。
典型地，这种修饰的MT-SP1多肽呈现对于补体蛋白C2增加的底物特异性。这种多肽的例子是含有相应于如下任何修饰的一或多个氨基酸修饰的那些MT-SP1多肽：MT-SP1多肽例如具有SEQ ID NO：253所示氨基酸序列的MT-SP1多肽或者SEQ ID NO：505所示其催化活性片段中的I136T/N164D/T166A/F184(A)L/D217V；I41F；I41F/A126T/V244G；D23E/I41F/T98P/T144I；I41F/L171F/V244G；H143R/Q175R；I41F/L171F；R230W；I41F/I154V/V244G；I141F/L52M/V129D/Q221(A)L；F99L；F97Y/I136V/Q192H/S201I；H71R/P131S/D217V；D217V；T65K/F93L/F97Y/D217V；I41T/P173S/Q209L；F97L/F234L；Q175R；N95K；和Y60(G)S。这种序列的例子是SEQ ID NO：589-609所示任何序列，及其含有所述突变且具有催化活性的连续氨基酸片段，例如SEQ ID NO：568-588所示任何序列。在一些实例中，本发明提供的变体MT-SP1多肽另外含有相应于MT-SP1多肽例如具有SEQ ID NO：253所示氨基酸序列的MT-SP1多肽或者SEQ IDNO：505所示其催化活性片段中的C122S的修饰。这种变体MT-SP1多肽的例子如SEQ ID NO：537-557所示，或者其含有所述突变且具有催化活性的连续氨基酸片段，例如SEQ ID NO：516-536所示任何序列。
特别地，本发明提供了具有如下氨基酸修饰的修饰的u-PA多肽：L73A/I89V；L155P；R72G/L155P；F132L/V160A；L73A/I89V/F30T；L73A/I89V/L155V；L73A/I89V/L155M；和L73A/I89V/F30L/L155M(基于胰凝乳蛋白酶编号)。
G.评估蛋白酶活性和特异性的方法
可以对在本发明提供的方法中选择的蛋白酶进行检测以确定在选择后所述蛋白酶是否保留催化效率且呈现出希望的底物特异性。可以使用来自扩增的展示源或者来自纯化的蛋白质的上清进行活性评估。例如，如上文所讨论，可以在用辅助噬菌体拯救噬菌体和噬菌体扩增后分析噬菌体上清。或者，蛋白酶活性可以从感染的细菌的细胞培养基或者其周质中直接测定。也可以确定纯化的选择的蛋白酶的蛋白酶活性。
可以使用已知的蛋白酶底物通过分析底物裂解而评估催化效率和/或底物特异性。例如，纤溶酶原的裂解可以在t-PA或u-PA用于所述选择方法中的情况下评估。在另一个实例中，可以使用由所述蛋白酶识别的肽底物。例如，是MT-SP1的自活化位点的RQAR(SEQ ID NO：513)可用于评估选择的MT-SP1蛋白酶的活性。在一个实施方案中，可以使用肽底物的荧光标记的四肽例如ACC-或AMC-四肽。另外，基于希望的靶底物的裂解序列设计的荧光肽底物可以用于评估活性，针对所述靶底物选择蛋白酶。
在一些实例中，可以在存在或不存在用于选择方法中的变体蛋白酶捕获多肽的条件下评估选择的蛋白酶针对已知肽底物的活性。典型地，进行这种活性评估以进一步选择在存在含有希望的靶底物裂解序列的蛋白酶捕获多肽的条件下被抑制的那些蛋白酶，从而优化选择的对于靶底物具有改良的选择性的蛋白酶。可以针对野生型或者模板蛋白酶和/或与在选择方法中鉴别的所有其它蛋白酶进行抑制作用对比。
可以将选择的蛋白酶的天然底物裂解的动力学分析与希望的靶底物的裂解分析进行对比以评估选择的蛋白酶对于靶序列的特异性。另外，可以评估抑制作用的二级速率常数(ki)以监测选择的蛋白酶对于底物的效率和反应性，所述底物例如所述选择方法中使用的蛋白酶捕获多肽或者其变体。实施例5举例说明了用于评估在本发明提供的方法中鉴别的突变体u-PA多肽的催化效率和反应性的各种测定方法。实施例10和实施例12举例说明了用于评估选择的MT-SP1噬菌体上清的催化效率的各种测定方法。实施例14举例说明了用于评估选择的纯化的变体MT-SP1蛋白酶的催化效率和反应性的各种测定方法。
在一个实例中，可以使用相应于希望的裂解序列的各个荧光肽底物分析选择的蛋白酶，例如分析根据匹配突变的蛋白酶捕获多肽的希望的特异性谱而选择的u-PA或MT-SP1蛋白酶。例如，分析可以裂解靶底物的任一或多个希望的裂解序列的修饰的蛋白酶的方法包括：(a)将肽荧光样品(含有希望的靶裂解序列)与蛋白酶接触，由此荧光部分在蛋白酶的作用下从肽底物序列中释放出来，从而产生荧光部分；(b)观测所述样品是否经历荧光的可检测变化，所述可检测变化表示样品中存在酶活性蛋白酶。在这种实例中，通过本领域已知方法将用于选择蛋白酶的希望的裂解序列制成荧光肽。在一个实施方案中，可以将各个肽裂解序列附着于荧光标记的底物，例如ACC或AMC荧光离去基团，可以确定所述荧光部分的释放以作为蛋白酶对于肽裂解序列的特异性的测量值。靶裂解序列的荧光增加速率可以例如通过使用荧光分光光度计测量。可以随着时间测量荧光增加速率。Michaelis-Menton动力学常数可以通过标准动力学方法确定。动力学常数kcat、Km和kcat/Km可以通过将底物浓度的倒数对底物裂解速度的倒数绘图并拟合Lineweaver-Burk方程(1/速度＝(Km/Vmax)(1/[S])+1/Vmax；其中Vmax＝[ET]kcat)而计算。所述二级速率常数或者特异性常数(kcat/Km)测量底物由特定蛋白酶切割底物程度的测量。例如，可以产生ACC-或AMC-四肽如Ac-RRAR-AMC、Ac-SLGR-AMC、Ac-SLGR-ACC、Ac-RQAR-ACC，将其与在本发明提供的方法中选择的蛋白酶保温，通过测定荧光部分的释放而评估蛋白酶的活性。所述四肽的选择依赖于被测定的希望的裂解序列，且可以根据经验确定。
测定溶液中的蛋白酶仅需要向荧光蛋白酶指示肽中加入一定量的蛋白酶原液并测量随后在吸收光谱中荧光的增加或激发带中的降低。所述溶液和荧光指示肽也可以组合并在优化所述蛋白酶的活性的“消化缓冲液”中测定。适于测定蛋白酶活性的缓冲液为本领域技术人员所熟知。通常，选择这样的缓冲液，即其pH相应于特定蛋白酶的最适pH。例如，特别适于测定弹性蛋白酶活性的缓冲液含有50mM磷酸钠、1mM EDTA，pH 8.9。所述测量在荧光计中最容易进行，所述荧光计是为荧光团提供“激发”光源，然后测量随后在特定波长发射的光的仪器。与无蛋白酶的对照指示溶液相比，提供了蛋白酶活性的测量。活性水平可以通过产生蛋白酶/指示剂组合的标准曲线而精确量化，在所述曲线中确定已知活性的蛋白酶溶液产生的荧光变化速率。
荧光化合物的检测可以使用荧光计完成，且可以通过本领域技术人员熟知的各种其它方法进行检测。因此，例如当荧光团在可见波长发射时，可以简便地通过目测响应光源激发的荧光进行检测。也可以通过图像分析系统进行检测，利用与数字化装置连接的摄像机或者其它图像获取系统进行检测。也可以在荧光显微镜下通过滤波器观测进行检测。该显微镜可以提供仅由操作者观测到的信号。或者，该信号可以记录在照相软片上或者使用视频系统记录。该信号也可以通过使用图像分析系统或者光度计简便地实时量化。
因此，例如，样品的蛋白酶活性的一种基本测定方法包括将样品悬浮于或者溶解于缓冲液(在对于被测定的特定蛋白酶最佳的pH)中，向缓冲液中加入荧光蛋白酶肽指示剂，并使用荧光分光光度计监测荧光中的变化，例如Harris et al.，(1998)J Biol Chem 273：27364示出。荧光分光光度计被设置为在荧光团的激发波长激发所述荧光团。荧光蛋白酶指示剂是蛋白酶的底物序列，由于蛋白酶裂解指示剂而导致荧光变化。
也可以测定选择的蛋白酶以确定当其在全长蛋白质情况中存在时将裂解希望的序列。在一个实例中，可以将纯化的靶蛋白即VEGFR2或补体蛋白C2在存在或不存在选择的蛋白酶的条件下保温，通过SDS-PAGE监测裂解情况，随后对蛋白质进行考马斯亮蓝染色，并使用光密度测定法分析裂解产物。蛋白酶对全长蛋白质的裂解特异性常数可以使用凝胶光密度测定法确定，以评估在存在蛋白酶的条件下保温的全长靶底物条带的光密度测定中随时间而发生的变化。另外，也可以使用本领域熟知的方法测定靶蛋白质的活性，以测定希望的靶蛋白质的活性，从而证实其功能已经由于裂解事件而被破坏。
在特定的实施方案中，选择的蛋白酶、典型为修饰的蛋白酶的特异性对比可用于确定选择的蛋白酶与野生型或模板蛋白酶相比是否呈现出改变的、例如增加的特异性。可以通过观测与野生型或模板蛋白酶相比，修饰的蛋白酶在指定活性裂解多少不同序列而测量蛋白酶对于靶底物的特异性。如果修饰的蛋白酶与野生型蛋白酶相比裂解较少的靶底物，则所述修饰的蛋白酶与野生型蛋白酶相比对于那些靶底物具有较高特异性。蛋白酶对于靶底物的特异性可以从靶底物与非靶底物(即蛋白酶的天然野生型底物序列)相比的裂解特异性常数确定。可以产生修饰的蛋白酶对于靶底物相对于非靶底物的特异性常数比率以确定蛋白酶裂解效率比率。修饰的蛋白酶与野生型或模板蛋白酶之间裂解效率比率的对比可用于评估对于靶底物的特异性的变化倍数。当与野生型或模板蛋白酶对于靶底物相对于非靶底物的特异性相对比时，特异性可以是至少2倍、至少4倍、至少5倍、至少6倍、至少7倍、8、9、10、20、30、40、50、60、70、80、90、100、200、300、400、500、600、700、800、900、1000倍或更多倍。
H.产生编码蛋白酶捕获多肽(即丝氨酸蛋白酶抑制蛋白)或者其变体或者蛋白酶/修饰的蛋白酶的核酸的方法
本文阐述的多肽，包括蛋白酶捕获多肽或者蛋白酶多肽或者其催化活性部分，包括修饰的u-PA多肽或者修饰的MT-SP1多肽，可以通过本领域熟知的蛋白质纯化和重组蛋白质表达方法获得。可以使用本领域技术人员已知的鉴别编码希望的基因的核酸的任何方法。本领域可利用的任何方法均可用于如从细胞或组织来源中获得编码希望的蛋白酶捕获多肽或蛋白酶蛋白的全长(即包含全部编码区)cDNA或基因组DNA克隆。修饰的多肽如变体蛋白酶捕获多肽或者选择的变体蛋白酶可以如本文所述例如通过定点诱变方法从野生型多肽工程化。
可以使用本领域已知的克隆和分离核酸分子的任何可利用的方法克隆或分离多肽。这种方法包括PCR扩增核酸和文库筛选，包括核酸杂交筛选、基于抗体的筛选和基于活性的筛选方法。
核酸扩增方法可用于分离编码希望的多肽的核酸分子，包括例如聚合酶链反应(PCR)方法。含有核酸的材料可以用作起始材料，从中可以分离希望的编码多肽的核酸分子。例如，来自健康和/或患病对象的DNA和mRNA制品、细胞提取物、组织提取物、流体样品(例如血液、血清、唾液)可用于扩增方法中。核酸文库也可以用作起始材料的来源。可以设计引物以扩增希望的多肽。例如，可以基于从中产生希望的多肽的表达的序列设计引物。引物可以基于多肽氨基酸序列的反向翻译(back-translation)设计。可以对通过扩增产生的核酸分子进行测序，并证实其编码希望的多肽。
其它核苷酸序列可以与多肽编码核酸分子连接，包括含有限制性核酸内切酶位点的接头序列，以将合成的基因克隆进载体中，所述载体例如是蛋白质表达载体或者设计为扩增核心蛋白编码DNA序列的载体。此外，限定功能性DNA元件的另外的核苷酸序列可以与多肽编码核酸分子可操纵地连接。这种序列的例子包括但不限于设计为便于胞内蛋白质表达的启动子序列和设计为便于蛋白质分泌的分泌序列。另外的核苷酸残基序列如限定蛋白质结合区的碱基序列也可以与蛋白酶编码核酸分子连接。这种区域包括但不限于便于蛋白酶吸收进特定靶细胞中的残基序列或者编码使得蛋白酶便于被吸收进特定靶细胞中的蛋白质的残基序列，或者另外改变合成基因的产物的药动学的残基序列。
另外，可以加入标记或者其它部分，例如以助于检测或者亲和性纯化所述多肽。例如，其它核苷酸序列例如指定表位标记或者其它可检测标记的碱基序列也可以与蛋白酶编码核酸分子或者丝氨酸蛋白酶抑制蛋白编码核酸分子或其变体连接。这种序列的例子是编码His标记(例如6×His，HHHHH；SEQ ID NO：496)或者Flag标记(DYKDDDDK；SEQ ID NO：495)的核酸序列。
然后可以将鉴别的和分离的核酸核酸插入适当的克隆载体中。可以使用本领域已知的大量载体-宿主系统。可能的载体包括但不限于质粒或修饰的病毒，但是载体系统必须与使用的宿主细胞相容。这种载体包括但不限于噬菌体例如λ噬菌体衍生物，或者质粒如pCMV4、pBR322或pUC质粒衍生物，或者Bluescript载体(Stratagene，La Jolla，CA)。插入克隆载体中可以例如通过将所述DNA片段连接在具有互补粘端的克隆载体中而完成。插入可以通过使用TOPO克隆载体(INVITROGEN，Carlsbad，CA)实现。如果用于使DNA片段化的互补限制位点不存在于克隆载体中，则可以对所述DNA分子的末端进行酶处理修饰。或者，通过将核苷酸序列(接头)连接在DNA末端上而可以产生任何希望的位点；这些连接的接头可以含有编码限制性核酸内切酶识别序列的经特异化学合成的寡核苷酸。在另一种方法中，裂解的载体和蛋白质基因可以通过同聚物加尾而修饰。重组分子可以通过例如转化、转染、感染、电穿孔和sonoporation等方式导入宿主细胞中，以便产生基因序列的许多拷贝。
在特定的实施方案中，用掺入了分离的蛋白质基因、cDNA或者合成的DNA序列的重组DNA分子转化宿主细胞，使得可以产生基因的多个拷贝。因此，通过生长转化体、从所述转化体中分离重组DNA分子、且当需要时从分离的重组DNA中回收插入的基因，可以获得大量的基因。
1.载体与细胞
为了重组表达一或多种希望的蛋白质，例如本文所述的任何蛋白质，可以将含有编码所述蛋白质的全部或部分核苷酸序列的核酸插入合适的表达载体中，所述载体即含有插入的蛋白质编码序列的转录和翻译必需的元件的载体。所述必需的转录和翻译信号也可以由蛋白酶基因和/或其两侧区域的天然启动子提供。
本发明还提供了含有编码所述蛋白酶或修饰的蛋白酶的核酸的载体。本发明还提供了含有所述载体的细胞。所述细胞包括真核细胞和原核细胞，所述载体是适用于此的任何载体。
本发明提供了含有所述载体的原核细胞和真核细胞，包括内皮细胞。这种细胞包括细菌细胞、酵母细胞、真菌细胞、古细菌(Archea)、植物细胞、昆虫细胞和动物细胞。所述细胞用于产生蛋白质，通过将所述细胞在使得所述细胞表达编码的蛋白质的条件下生长，并回收表达的蛋白质进行。对于本发明而言，例如所述蛋白酶可以被分泌进培养基中。
在一个实施方案中，提供了含有核苷酸序列的载体，所述核苷酸序列编码具有蛋白酶活性的多肽，例如编码本发明提供的任何u-PA变体多肽，且含有全部或部分蛋白酶结构域或者其多个拷贝。本发明还提供了含有核苷酸序列的载体，所述核苷酸序列编码蛋白酶结构域和蛋白酶蛋白的其它部分，直至并包括全长蛋白酶蛋白，以及其多个拷贝。载体可以针对细胞中修饰的蛋白酶蛋白或者其蛋白酶结构域的表达而选择，或者所述蛋白酶蛋白表达为分泌蛋白。当蛋白酶结构域被表达时，所述核酸与编码分泌信号的核酸连接，所述分泌信号例如是酿酒酵母(Saccharomyces cerevisiae)交配因子信号序列或者其一部分，或者天然信号序列。
许多宿主-载体系统可用于表达蛋白质编码序列。这些包括但不限于用病毒(例如痘苗病毒、腺病毒及其它病毒)感染的哺乳动物细胞系统；用病毒(例如杆状病毒)感染的昆虫细胞系统；微生物如含有酵母载体的酵母；或者用噬菌体、DNA、质粒DNA或者粘粒DNA转化的细菌。载体的表达元件根据其强度和特异性而有所不同。根据使用的宿主-载体系统，可以使用任何合适的转录和翻译元件。
可以使用本领域技术人员已知的将DNA片段插入载体中的任何方法构建含有嵌合基因的表达载体，所述嵌合基因含有合适的转录/翻译控制信号和蛋白酶编码序列。这些方法可包括体外重组DNA和合成技术及体内重组方法(遗传重组)。编码蛋白质或其结构域、衍生物、片段或同系物的核酸序列的表达可以由另一个核酸序列调节，由此所述基因或其片段在用所述重组DNA分子转化的宿主中表达。例如，蛋白质的表达可以由本领域已知的任何启动子/增强子控制。在特定的实施方案中，启动子对于希望的蛋白质的基因不是天然的。可以使用的启动子包括但不限于SV40早期启动子(Bernoist and Chambon，Nature 290：304-310(1981))、包含于Rous肉瘤病毒的3’长末端重复中的启动子(Yamamoto et al.Cell 22：787-797(1980))、疱疹胸苷酸激酶启动子(Wagner et al.，Proc.Natl.Acad.Sci.USA 78：1441-1445(1981))、金属硫蛋白基因的调节序列(Brinster et al.，Nature 296：39-42(1982))；原核表达载体如β-内酰胺酶启动子(Jay et al.，(1981)Proc.Natl.Acad.Sci.USA 78：5543)或者tac启动子(DeBoer et al.，Proc.Natl.Acad.Sci.USA80：21-25(1983))；也见“Useful Proteins from Recombinant Bacteria”：inScientific American 242：79-94(1980))所述；含有胭脂氨酸合成酶启动子的植物表达载体(Herrar-Estrella et al.，Nature 303：209-213(1984))或者花椰菜花叶病毒35S RNA启动子(Garder et al.，Nucleic Acids Res.9：2871(1981))，和光合作用酶核酮糖二磷酸羧化酶的启动子(Herrera-Estrella et al.，Nature3l0：115-120(1984))；来自酵母和其它真菌的启动子元件，如Gal4启动子、醇脱氢酶启动子、磷酸甘油激酶启动子、碱性磷酸酶启动子，和呈现组织特异性且已经用于转基因动物中的如下动物转录控制区：弹性蛋白酶I基因控制区，其在胰腺腺泡细胞中是活性的(Swift et al.，Cell 38：639-646(1984)；Ornitz et al.，Cold Spring Harbor Symp.Quant.Biol.50：399-409(1986)；MacDonald，Hepatology 7：425-515(1987))；胰岛素基因控制区，其在胰腺β细胞中是活性的(Hanahan et al.，Nature 3l5：115-122(1985))，免疫球蛋白基因控制区，其在淋巴细胞中是活性的(Grosschedl et al.，Cell38：647-658(1984)；Adams et al.，Nature 318：533-538(1985)；Alexander et al.，Mol.Cell Biol.7：1436-1444(1987))，小鼠乳腺肿瘤病毒控制区，其在睾丸、乳腺、淋巴细胞和肥大细胞中是活性的(Leder et al.，Cell 45：485-495(1986))，白蛋白基因控制区，其在肝脏中是活性的(Pinckert et al.，Genes and Devel.1：268-276(1987))，α-胎蛋白基因控制区，其在肝脏中是活性(Krumlauf et al.，Mol.Cell.Biol.5：1639-1648(1985)；Hammer et al.，Science 235：53-58 1987))，α-1抗胰蛋白酶基因控制区，其在肝脏中是活性的(Kelsey et al.，Genes andDevel.1：161-171(1987))，β球蛋白基因控制区，其在髓样细胞中是活性的(Mogram et al.，Nature 315：338-340(1985)；Kollias et al.，Cell 46：89-94(1986))，髓磷脂碱性蛋白基因控制区，其在脑的少突胶质细胞中是活性的(Readhead et al.，Cell 48：703-712(1987))，肌球蛋白轻链-2基因控制区，其在骨骼肌中是活性的(Sani，Nature 314：283-286(1985))，促性腺激素释放激素基因控制区，其在下丘脑的促性腺激素细胞中是活性的(Mason et al.，Science234：1372-1378(1986))。
在特异的实施方案中，使用这样的载体，所述载体含有与编码希望的蛋白质或其结构域、片段、衍生物或同系物的核酸可操纵地连接的启动子，一或多个复制起点，任选一或多个可选择的标记(例如抗生素抗性基因)。例如，表达蛋白酶蛋白的蛋白酶结构域的载体和系统包括熟知的毕赤酵母(Pichia)载体(例如可得自Invitrogen，San Diego，CA)，特别是为分泌编码的蛋白质而设计的那些载体。转化大肠杆菌细胞的质粒载体的例子包括例如pQE表达载体(可得自Qiagen，Valencia，CA；也见Qiagen描述该系统的公开出版的文献所述)。pQE载体具有噬菌体T5启动子(由大肠杆菌RNA聚合酶识别)和双lac操纵基因阻抑模块以提供重组蛋白在大肠杆菌中的紧密调节的高水平表达，合成的核糖体结合位点(RBS II)以有效翻译，6×His标记编码序列，t0和T1转录终止子，ColE1复制起点，和赋予氨苄青霉素抗性的β-内酰胺酶基因。pQE载体允许在重组蛋白的N或C末端放置6×His标记。这种质粒包括pQE 32、pQE 30和pQE 31，其为所有三个读框提供多克隆位点，且供给N末端6×His标记的蛋白质的表达。转化大肠杆菌细胞的其它举例的质粒载体包括例如pET表达载体(见美国专利4,952,496；可得自NOVAGEN，Madison，WI；也见Novagen描述该系统的公开出版的文献所述)。这种质粒包括pET 11a，其含有T7lac启动子、T7终止子、可诱导的大肠杆菌lac操纵基因，和lac阻抑基因；pET 12a-c，其含有T7启动子、T7终止子和大肠杆菌ompT分泌信号；pET 15b和pET19b(NOVAGEN，Madison，WI)，其含有His-TagTM前导序列以用于在His柱中纯化，允许在经过该柱纯化后裂解的凝血酶裂解位点，T7-lac启动子区和T7终止子。
2.表达
蛋白质，如本文阐述的任何蛋白质，包括任何蛋白酶捕获多肽或其变体、或者选择的蛋白酶或其催化活性部分，可以通过本领域技术人员已知的任何方法产生，包括体内和体外方法。希望的蛋白质可以在适于产生需要量和需要形式的蛋白质的任何生物体中表达，所述蛋白质例如为给药和治疗所需的。表达宿主包括原核生物和真核生物，如大肠杆菌、酵母、植物、昆虫细胞、哺乳动物细胞包括人细胞系和转基因动物。表达宿主在蛋白质产生水平以及表达的蛋白质上存在的翻译后修饰类型方面可有所不同。表达宿主可以基于这些及其它因素加以选择，例如行政审批和安全性问题、生产成本和纯化的必要性和方法。
本领域技术人员可利用且已知许多表达载体，这些表达载体可用于表达蛋白质。宿主表达系统的选择影响表达载体的选择。通常表达载体可包括转录启动子及任选地增强子、翻译信号、转录和翻译终止信号。用于稳定转化的表达载体典型具有可选择的标记，所述可选择的标记使得可以选择和维持转化的细胞。在一些情况中，复制起点可用于扩增载体的拷贝数。
蛋白质，例如本发明提供的任何变体蛋白酶或者任何蛋白酶捕获多肽或其变体，也可以用作或者表达为融合蛋白。例如，蛋白酶融合可以通过向蛋白酶中加入另外的功能性而产生。蛋白酶融合蛋白的例子包括但不限于信号序列、标记如用于定位的标记例如his6标记或myc标记、或者用于纯化的标记例如GST融合体和指导蛋白酶分泌和/或膜结合的序列的融合体。
在一个实施方案中，蛋白酶可以活性形式表达。在另一个实施方案中，蛋白酶以无活性的酶原形式表达。
a.原核细胞
原核生物，特别是大肠杆菌，提供了产生大量蛋白质的系统。大肠杆菌的转化是本领域技术人员熟知的简便且快速的技术。大肠杆菌的表达载体可以含有可诱导的启动子，这种启动子可用于诱导高水平的蛋白质表达及表达对于宿主细胞呈现一些毒性的蛋白质。可诱导的启动子的例子包括lac启动子、trp启动子、杂种tac启动子、T7和SP6RNA启动子和温度调节的λPL启动子。
蛋白质，如本发明提供的任何蛋白质，可以在大肠杆菌的细胞质环境中表达。细胞质是一种还原环境，对于一些分子而言可以导致不溶的包含体形成。还原剂如二硫苏糖醇和β-巯基乙醇及变性剂如盐酸胍和脲可用于溶解所述蛋白质。另一种方法是蛋白质在细菌壁膜间隙中表达，这样提供了氧化环境和侣伴蛋白样和二硫化物异构酶，且可以导致可溶蛋白质的产生。典型地，将前导序列与被表达的蛋白质融合，指导所述蛋白质至周质。然后通过周质内部的信号肽酶除去该前导序列。靶向周质的前导序列的例子包括来自果胶酸裂合酶基因的pelB前导序列和衍生自碱性磷酸酶基因的前导序列。在一些情况中，周质表达使得被表达的蛋白质渗漏入培养基中。蛋白质的分泌使得可以从培养上清中快速和简便地纯化。非分泌的蛋白质可以通过渗透性细胞溶解而得自周质。与细胞质表达相似，在一些情况中蛋白质可以变为不溶的且可以使用变性剂和还原剂以便于溶解和再折叠。诱导和生长的温度也可以影响表达水平和溶解性，典型使用温度介于25℃与37℃之间。典型地，细菌产生非糖基化的蛋白质。因此，如果蛋白质需要糖基化以发挥其功能，则可以在从宿主细胞中纯化之后在体外加入糖基化。
b.酵母细胞
酵母如酿酒酵母(Saccharomyces cerevisae)、粟酒裂殖酵母(Schizosaccharomyces pombe)、解脂耶氏酵母(Yarrowia lipolytica)、乳酸克鲁维酵母(Kluyveromyces lactis)和巴斯德毕赤酵母(Pichia pastoris)是熟知的酵母表达宿主，可用于产生蛋白质，如本发明所述任何蛋白质。酵母可以用附加型复制载体转化或者通过同源重组进行稳定染色体整合。典型地，可诱导的启动子用于调节基因表达。这种启动子的例子包括GAL1，GAL7和GAL5及金属硫蛋白启动子如CUP1、AOX1或者其它毕赤酵母或其它酵母启动子。表达载体通常包括可选择的标记如LEU2、TRP1、HIS3和URA3，以选择和维持转化的DNA。在酵母中表达的蛋白质通常是可溶的。与侣伴蛋白如Bip和蛋白质二硫化物异构酶的共表达可以改良表达水平和溶解性。此外，在酵母中表达的蛋白质可以使用分泌信号肽融合体指导分泌，所述融合体如来自酿酒酵母的酵母交配型α-因子分泌信号和与酵母细胞表面蛋白如Aga2p交配粘附受体或者Arxula adeninivorans葡糖淀粉酶的融合体。蛋白酶如Kex-2的裂解位点可以被工程化以当被表达的多肽退出分泌途径时从其中除去融合的序列。酵母也能在Asn-X-Ser/Thr基序糖基化。
c.昆虫细胞
昆虫细胞特别是使用杆状病毒表达的昆虫细胞可用于表达多肽如修饰的蛋白酶或者修饰的蛋白酶捕获多肽。昆虫细胞表达高水平的蛋白质且大多数翻译后修饰能为更高等的真核生物所利用。杆状病毒具有限制性宿主范围，改善了真核表达的安全性并减少了行政审批方面的考虑。典型的表达载体使用启动子以高水平表达如杆状病毒的多角体蛋白启动子。常用的杆状病毒系统包括杆状病毒例如苜蓿银纹夜蛾(Autographa californica)核型多角体病毒(AcNPV)和家蚕(bombyx mori)核型多角体病毒(BmNPV)及昆虫细胞系如衍生自草地贪夜蛾(Spodoptera frugiperda)的Sf9、美洲粘虫(Pseudaletia unipuncta)(A7S)和君主斑蝶(Danaus plexippus)(DpN1)。为了高水平表达，将待被表达的分子的核苷酸序列融合在病毒的多角体蛋白起始密码子的立即下游。哺乳动物分泌信号在昆虫细胞中被精确加工且可以用于使表达的蛋白质分泌进培养基中。另外，细胞系美洲粘虫(A7S)和君主斑蝶(DpN1)产生具有与哺乳动物细胞系统相似的糖基化模式的蛋白质。
在昆虫细胞中另一种表达系统是使用稳定转化的细胞。细胞系如Schnieder 2(S2)和Kc细胞(黑腹果蝇(Drosophila melanogaster))及C7细胞(白纹伊蚊(Aedes albopictus))可用于表达。果蝇金属硫蛋白启动子在存在重金属镉或铜诱导条件下可用于诱导高水平的表达。表达载体典型地通过使用可选择的标记如新霉素和潮霉素维持。
d.哺乳动物细胞
哺乳动物表达系统可用于表达蛋白质，包括修饰的蛋白酶或其催化活性部分，或者蛋白酶捕获多肽或其变体。表达构建体可以通过如下方式转移至哺乳动物细胞中，例如通过病毒感染如腺病毒感染或者通过直接DNA转移如脂质体、磷酸钙、DEAE-葡聚糖转移及通过物理方式如电穿孔和显微注射方式转移。用于哺乳动物的表达载体典型包括一个mRNA帽位点、TATA盒、翻译起始序列(Kozak共有序列)和聚腺苷酸化元件。这种载体通常包括转录启动子-增强子以高水平表达，例如SV40启动子-增强子、人巨细胞病毒(CMV)启动子和Rous肉瘤病毒(RSV)的长末端重复。这些启动子-增强子在许多细胞类型中是活性的。组织和细胞类型启动子及增强子区域也可用于表达。启动子/增强子区域的例子包括但不限于如下基因的那些启动子/增强子：弹性蛋白酶I、胰岛素、免疫球蛋白、小鼠乳腺肿瘤病毒、白蛋白、α-胎蛋白、α1抗胰蛋白酶、β球蛋白、髓磷脂碱性蛋白、肌球蛋白轻链2，和促性腺激素释放激素基因对照(control)。可选择的标记可用于选择和维持具有所述表达构建体的细胞。可选择的标记基因的例子包括但不限于潮霉素B磷酸转移酶、腺苷脱氨酶、黄嘌呤-鸟嘌呤磷酸核糖转移酶、氨基糖苷磷酸转移酶、二氢叶酸还原酶和胸苷激酶。与细胞表面信号分子如TCR-ζ和FcεRI-γ的融合可以指导蛋白质在细胞表面上以活性状态表达。
许多细胞系可用于哺乳动物表达，包括小鼠、大鼠、人、猴、鸡与仓鼠细胞。细胞系的例子包括但不限于CHO、Balb/3T3、HeLa、MT2、小鼠NS0(非分泌)及其它骨髓瘤细胞系、杂交瘤和异杂交瘤(heterohybridoma)细胞系、淋巴细胞、成纤维细胞、Sp2/0、COS、NIH3T3、HEK293、293S、2B8和HKB细胞。细胞系也可以用于适合无血清培养基，便于分泌的蛋白质从细胞培养基中纯化。一个这样的例子是无血清EBNA-1细胞系(Pham etal.，(2003)Biotechnol.Bioeng.84：332-42)。
e.植物
转基因植物细胞和植物可用于表达蛋白质，如本发明描述的任何蛋白质。使用直接DNA转移及用农杆菌介导的转化方法将表达构建体典型地转移至植物中，所述直接DNA转移例如是通过微粒轰击和PEG介导的转移进原生质体中。表达载体可包括启动子和增强子序列、转录终止元件和翻译控制元件。表达载体和转化技术在双子叶植物宿主如拟南芥(Arabidopsis和烟草与单子叶植物宿主如玉米和水稻之间通常有所不同。用于表达的植物启动子的例子包括花椰菜花叶病毒启动子、胭脂氨酸合酶启动子、核糖二磷酸羧化酶启动子和遍在蛋白及UBQ3启动子。可选择的标记如潮霉素、磷酸甘露糖异构酶和新霉素磷酸转移酶通常用于促进转化的细胞的选择和维持。转化的植物细胞可以作为细胞、聚集体(愈伤组织)维持培养或者再生为全植柱。转基因植物细胞也可包括藻类，其工程化为产生蛋白酶或修饰的蛋白酶(见例如Mayfield et al.(2003)PNAS 100：438-442)。由于植物具有不同于哺乳动物细胞的糖基化模式，因此可以影响在这些宿主中产生的蛋白质的选择。
3.纯化技术
来自宿主细胞的多肽、包括蛋白酶多肽或其它蛋白质的纯化方法将依赖于选择的宿主细胞和表达系统。对于分泌的分子，蛋白质通常在除去细胞之后从培养基中纯化。对于胞内表达，可以使细胞溶解，然后从提取物中纯化蛋白质。当转基因生物如转基因植物和动物用于表达时，组织或器官可以用作起始材料以产生溶解的细胞提取物。此外，转基因动物生产可包括在乳汁或蛋中产生多肽，可以将其收集，且如果需要时，使用本领域的标准方法提取蛋白质并进行进一步纯化。
在一个实例中，蛋白酶可以无活性形式(酶原形式)被表达和纯化或者通过自身催化除去前区(proregion)，表达的蛋白酶可以被纯化为活性形式如双链形式。典型地，自主激活在纯化过程期间发生，例如在室温保温24-72小时期间发生。激活的速度和程度依赖于蛋白质浓度和特异修饰的蛋白酶，由此例如更稀释的样品可需要在室温保温更长的时间。激活可以通过SDS-PAGE(例如3千道尔顿移位)及通过酶活性(荧光底物的裂解)进行监测。典型地，在纯化前使蛋白酶达到＞75％的激活。
蛋白质如蛋白酶或者蛋白酶捕获多肽可以使用本领域已知的标准蛋白质纯化方法纯化，所述方法包括但不限于SDS-PAGE、大小分级分离和大小排阻层析、硫酸铵沉淀和离子交换层析如阴离子交换层析。亲和纯化技术也可以用于改良制备物的效率和纯度。例如，结合蛋白酶或蛋白酶捕获多肽的抗体、受体及其它分子可用于亲和纯化中。表达构建体也可以工程化为向蛋白质中加入亲和性标记如myc表位、GST融合体或者His6，并且分别用myc抗体、谷胱甘肽树脂和Ni-树脂亲和性纯化。纯度可以通过本领域已知的任何方法评估，包括使用凝胶电泳和染色及分光光度法评估。
4.融合蛋白
本发明还提供了含有本发明提供的变体蛋白酶和一或多个其它多肽的融合蛋白。本发明提供了含有这种融合蛋白的药物组合物，所述药物组合物配制为通过合适途径给予。融合蛋白是通过修饰的蛋白酶与另一多肽以任何顺序连接而形成，所述另一多肽例如是抗体或其片段、生长因子、受体、配体及其它这种物质以便于蛋白酶的纯化、通过指导所述蛋白酶至靶细胞或组织而改变蛋白酶的药动学性质和/或增加蛋白酶的表达或分泌。在蛋白酶融合蛋白中，蛋白酶多肽可相应于蛋白酶蛋白的全部或者其催化活性部分。在一些实施方案中，所述蛋白酶或其催化活性部分是修饰的蛋白酶。本发明提供的融合蛋白基本保留其对于任一或多个希望的靶底物的所有特异性和/或选择性。通常蛋白酶融合多肽与非融合蛋白酶相比保留至少大约30％、40％、50％、60％、70％、80％、85％、90％或95％的底物特异性和/或选择性，包括与非融合蛋白酶相比保留96％、97％、98％、99％或更高的底物特异性。
蛋白酶多肽与另一多肽的连接可以直接连接或者通过接头间接连接。在一个实例中，连接可以是通过化学键连接，如通过异双功能试剂或者硫醇键或者其它这种键连接。蛋白酶与另一多肽的融合可以是与蛋白酶多肽的N或C末端融合。可用于与本发明提供的蛋白酶的融合蛋白中的多肽的非限制性实例包括例如GST(谷胱甘肽S-转移酶)多肽、免疫球蛋白的Fc结构域或者异源信号序列。所述融合蛋白可含有另外的成分，如大肠杆菌麦芽糖结合蛋白(MBP)，其有助于细胞摄取蛋白质(见国际PCT申请No.WO01/32711)。
蛋白酶融合蛋白可以通过标准重组方法产生。例如，编码不同多肽序列的DNA片段可以根据常规技术符合读框地连接在一起，例如通过应用平末端或者交错末端进行连接、限制酶消化以提供合适的末端，适当充填粘端，经碱性磷酸酶处理以避免不希望的连接，并通过酶连接。在另一个实施方案中，融合基因可以通过常规方法合成，包括利用自动DNA合成仪。或者，可以使用锚引物对基因片段进行PCR扩增，所述引物在两个连续基因片段之间形成互补突出端，随后可退火并再扩增以产生嵌合基因序列(见例如Ausubel et al.(eds.)CURRENT PROTOCOLS IN MOLECULARBIOLOGY，John Wiley & Sons，1992)。此外，可以商购已编码融合部分(例如GST多肽)的许多表达载体。可以将蛋白酶编码核酸克隆进这种表达载体中，由此所述融合部分与蛋白酶蛋白符合读框地连接。
5.核苷酸序列
本发明提供了编码修饰的蛋白酶的核酸分子。核酸分子包括任何编码的蛋白酶或其催化活性部分的等位基因变体或者剪接变体。在一个实施方案中，本发明提供的核酸分子与至少70％的全长的编码野生型蛋白酶或者其催化活性部分的任何核酸具有至少50、60、65、70、75、80、85、90、91、92、93、94、95或99％的序列相同性或者在中等或高度严格条件下与其杂交。在另一个实施方案中，核酸分子可包括具有例如本发明提供的任何蛋白酶或其催化活性部分的简并密码子序列的那些核酸分子。
本发明提供了与启动子可操纵地连接以在哺乳动物中表达的核酸分子或者含有核酸分子的催化活性部分的融合蛋白，所述启动子如可诱导的启动子。这种启动子包括但不限于CMV和SV40启动子；腺病毒启动子如E2基因启动子，其应答HPV E7癌蛋白；PV启动子，如PBV p89启动子，其应答PV E2蛋白；及由HIV或PV或癌基因激活的其它启动子。
本发明提供的修饰的蛋白酶也可以在基因转移载体中输送至细胞中。所述转移载体也可以编码另外的其它治疗剂以治疗给予所述蛋白酶的疾病或病症如凝血功能障碍或者癌症。编码蛋白酶的转移载体可以通过给予对象所述核酸而全身性应用。例如，转移载体可以是病毒载体如腺病毒载体。编码蛋白酶的载体也可以掺入干细胞中，这种干细胞例如通过在治疗部位移植(transplanting)或者移入(engrafting)所述干细胞而给予对象。例如，间充质干细胞(MSC)可以工程化为表达蛋白酶且这种MSC被移入在肿瘤部位进行治疗。
I.选择的蛋白酶多肽的制备、配制和给予
1.组合物与输送
可以配制选择的蛋白酶例如选择的突变体u-PA多肽的组合物以通过本领域技术人员已知的任何途径给予，包括肌内、静脉内、皮内、腹膜内注射、皮下、硬膜外、鼻、口服、直肠、局部、吸入、口腔(例如舌下)和经表皮给予或者任何途径给予。选择的蛋白酶可以通过任何常规途径给予，例如通过输注或者推注给予、通过上皮层或皮肤粘膜层(例如口腔粘膜、直肠和肠道粘膜等)且可以与其它生物活性物质相继、间歇或于相同组合物中一起给予。根据治疗位置可以局部、表面或者全身性给予。局部给予需要治疗的部位可以通过例如但不限于在手术期间局部输注而实现，表面应用例如在手术后与创伤敷料共同应用、通过注射、利用导管、利用栓剂或者利用植入物应用。给予也可包括通过控制释放系统给予，该系统包括控制释放制剂与控制释放装置例如泵。在任何指定情况中最合适的途径根据多种因素而定，例如疾病的性质、疾病的进展、疾病的严重性和使用的特定组合物。
本领域已知许多输送系统可用于给予选择的蛋白酶，这些系统例如但不限于在脂质体中的包囊、微粒、微囊、能表达所述化合物的重组细胞、受体介导的胞吞作用，编码选择的蛋白酶的核酸分子的输送如逆转录病毒输送系统。
可以制备含有选择的蛋白酶的药物组合物。通常药物可接受的组合物是根据用于动物和人体的公认药典，鉴于管理机构或者其它机构的许可而制备的。药物组合物可包括载体如稀释剂、佐剂、赋形剂或者利用其给予同工型的运载体。这种药物载体可以是无菌液体，如水和油，包括源自石油、动物、植物或者合成的那些，例如花生油、大豆油、矿物油和芝麻油。当药物组合物经静脉内给予时，水是典型的载体。盐水溶液和葡萄糖水溶液及甘油溶液也可以用作液体载体，特别是对于可注射的溶液。组合物除了活性成分之外，还可以含有：稀释剂，如乳糖、蔗糖、磷酸二钙或者羧甲基纤维素；滑润剂，如硬脂酸镁、硬脂酸钙和滑石；结合剂，如淀粉、天然树胶如阿拉伯胶、明胶、葡萄糖、糖蜜、聚乙烯吡咯烷酮、纤维素及其衍生物、聚维酮、交聚维酮，及本领域技术人员已知的其它这种结合剂。合适的药物赋形剂包括淀粉、葡萄糖、乳糖、蔗糖、明胶、麦芽、水稻、面粉、白垩、硅胶、硬脂酸钠、单硬脂酸甘油酯、滑石、氯化钠、脱脂奶粉、甘油、丙烯、乙二醇、水和乙醇。如果需要，组合物还可以含有少量的湿润剂或者乳化剂、或者pH缓冲剂，例如乙酸盐、柠檬酸钠、环式糊精衍生物、失水山梨醇单月桂酸酯、三乙醇胺乙酸钠、油酸三乙醇胺及其它这种物质。这些组合物可以是溶液、悬浮液、乳状液、片剂、丸剂、胶囊、粉末和持续释放制剂形式。组合物可以与传统结合剂和载体如甘油三酯配制为栓剂。口服制剂可包括标准载体如药物级的甘露醇、乳糖、淀粉、硬脂酸镁、糖精钠、纤维素、碳酸镁及其它这种物质。合适的药物载体的例子在E.W.Martin所著″Remington′s Pharmaceutical Sciences″中描述。这种组合物可含有通常为纯化形式的治疗有效量的化合物以及适量的载体以提供正确给予患者的形式。制剂应适于给予模式。
本发明提供了以单位剂量形式给予人和动物的制剂，如片剂、胶囊、丸剂、粉末、颗粒、无菌胃肠外溶液或悬浮液、口服溶液或悬浮液、含有合适数量化合物或其药物可接受衍生物的油水乳状液。药物治疗活性化合物及其衍生物典型以单位剂量形式或者多剂量形式配制和给予。每个单位剂量含有足以产生希望的治疗作用的预定数量的治疗活性化合物以及必需的药物载体、运载体(vehicle)或稀释剂。单位剂量形式的例子包括安瓿和注射器及单独包装的片剂或胶囊。单位剂量形式可以部分或者多次给予。多剂量形式是包装在一个容器中的多个相同单位剂量形式，以单独的单位剂量形式给予。多剂量形式的例子包括小瓶(vial)、片剂或胶囊瓶或者品脱或加仑瓶。因此，多剂量形式是未单独包装的多个单位剂量。
可以制备含有0.005％至100％范围的活性成分的剂量形式或组合物，用非毒性载体平衡。对于口服，药物组合物可采取例如片剂或胶囊形式，通过常规方式利用药物可接受赋形剂制备，所述赋形剂例如是结合剂(例如预胶化的玉米淀粉、聚乙烯吡咯烷酮或者羟丙基甲基纤维素)；充填剂(例如乳糖、微晶纤维素或者磷酸氢钙)；滑润剂(例如硬脂酸镁、滑石或硅石)；分解剂(例如马铃薯淀粉或者羟乙酸淀粉钠)；或者湿润剂(例如十二烷基硫酸钠)。可以通过本领域熟知的方法对片剂进行包被。
药物制备物也可以是液体形式，例如溶液、糖浆或者悬浮液形式，或者可以是在使用之前与水或者其它合适载体重建的药品。这种液体制备物可以通过常规方式利用药物可接受的添加剂制备，所述添加剂如悬浮剂(例如山梨糖醇糖浆、纤维素衍生物或者氢化可食用脂肪)；乳化剂(例如卵磷脂或阿拉伯胶)；非水载体(例如杏仁油、油酯(oily ester)或者分馏的植物油)；和防腐剂(例如甲基或丙基-p-羟基苯甲酸酯或者山梨酸)。
适于直肠给药的制剂可以单位剂量栓剂提供。这些制剂可以通过将活性化合物与一或多种常规固体载体例如可可油混合、然后定形所得混合物而制备。
适于皮肤或眼的表面应用的制剂包括软膏、乳膏、洗剂、糊剂、凝胶、喷雾剂、气雾剂和油剂。举例的载体包括凡士林、羊毛脂、聚乙二醇、醇及两或多种这些物质的组合。表面应用制剂也可以含有占重量0.05至15、20、25％的增稠剂，所述增稠剂选自羟丙基甲基纤维素、甲基纤维素、聚乙烯吡咯烷酮、聚乙烯醇、聚亚烷基二醇、聚/羟烷基，(甲)丙烯酸脂或者聚(甲)丙烯酰胺。表面制剂通常以滴剂或者软膏用于结膜囊。其也可以用于冲洗或润滑眼、面部窦(facial sinuses)和外耳道。其也可以被注入眼的前房及其它部位。液体状态的表面制剂也可以条(strip)或接触镜形式存在于亲水性体型聚合物基质中，从中释放所述活性成分。
对于经吸入给予，使用的化合物可以气雾剂喷雾形式从加压包或喷雾器中利用合适的推进剂输送，所述推进剂例如二氯二氟甲烷、三氯氟甲烷、二氯四氟乙烷、二氧化碳或者其它合适气体。在加压气雾剂的情况中，剂量单位可以通过计量所述的阀门决定。用于吸入器或吹入器中的胶囊和药筒例如明胶可以配制为含有所述化合物与合适的粉末基质如乳糖或淀粉的粉末混合物。
适于口腔(舌下)给予的制剂包括例如锭剂(lozenge)，其含有在调味基质中的活性化合物，所述基质通常为蔗糖和阿拉伯胶或者黄芪胶；软锭剂(pastille)，其含有在惰性基质中的化合物，所述基质例如是明胶和甘油或者蔗糖和阿拉伯胶。
选择的蛋白酶的药物组合物可以配制为通过注射胃肠外给予，例如通过推注或者持续输注给予。注射制剂可以单位剂量形式存在，例如在加入了防腐剂的安瓿或者在多剂量容器中。所述组合物可以是于油状或者水运载体中的悬浮液、溶液或者乳状液，且可以含有配方剂(formulatory agent)，如悬浮剂、稳定剂和/或分散剂。或者，活性成分可以是粉末形式，在使用之前与合适的运载体例如无菌无热原水或者其它溶剂重建。
本发明提供了适于经皮给予的制剂。该制剂可以是任何适当形式例如适合与受者的表皮保持长期紧密接触的分立的贴剂(patch)。这种贴剂含有于任选缓冲的水溶液中的活性化合物，所述溶液的浓度例如是就活性化合物而言0.1-0.2M。适于经皮给予的制剂也可以通过离子电渗法给予(见例如Pharmaceutical Research 3(6)，318(1986))，典型采取任选缓冲的活性化合物的水溶液形式。
药物组合物也可以通过控制释放制剂和/或给予装置给予(见例如美国专利No.3,536,809、3,598,123、3,630,200、3,845,770、3,847,770、3,916,899、4,008,719、4,687,610、4,769,027、5,059,595、5,073,543、5,120,548、5,354,566、5,591,767、5,639,476、5,674,533和5,733,566)。
在某些实施方案中，脂质体和/或纳米颗粒也可以用于给予选择的蛋白酶。脂质体是从磷脂形成的，其在水介质中分散且自发形成多层同心双层小泡形式(也称作多层小泡(MLV))。MLV通常直径为25nm至4μm。对MLV进行超声处理导致小单层小泡(SUV)形成，其直径为200-500埃，在中心含有水溶液。
当磷脂分散于水中时，根据脂质与水的摩尔比率可以形成不同于脂质体的多种结构。在低比率时，形成脂质体。脂质体的物理特性依赖于pH、离子强度和二价阳离子的存在。脂质体可以示出对于离子和极性物质的低通透性，但是在温度升高时经历相变而明显改变其通透性。所述相变包括从所谓的密集的、有序结构的凝胶状态向所谓的松散的、无序(1ess-ordered)结构的流体状态转变。这在特征相变温度发生，并导致对于离子、糖和药物的通透性增加。
脂质体与细胞通过不同机制相互作用：通过网状内皮系统的吞噬细胞如巨噬细胞和嗜中性粒细胞的胞吞作用；吸附于细胞表面，通过非特异性弱疏水力或静电力或者通过与细胞表面成分的特异性相互作用而吸附；与浆细胞膜融合，通过脂质体的脂质双层插入质膜中，同时脂质体内容物释放进细胞质中；通过脂质体脂质转移至细胞膜或者亚细胞膜，反之亦然，而与脂质内容物无任何关联。改变脂质体配方可以改变运行机制，尽管同时可运行一个以上的机制。纳米胶囊(nanocapsule)通常可以稳定和可再生方式诱捕(entrap)化合物。为了避免由于胞内聚合过载的副作用，应使用能在体内降解的聚合物设计这种超细粒子(大小为大约0.1μm)。符合这些需要的可生物降解的聚烷基-氰基丙烯酸酯纳米粒子预期用于本发明中，且这种粒子可以很容易地产生。
给予方法可用于降低选择的蛋白酶暴露于降解过程，如蛋白酶解及由于抗原性和免疫原性应答引起的免疫干预。这种方法的例子包括在治疗部位局部给予。已经报道了聚乙二醇化治疗剂增加对于蛋白酶解的抗性、增加血浆半衰期、及降低抗原性和免疫原性。聚乙二醇化方法学的例子为本领域所已知(见例如Lu and Felix，Int.J.Peptide Protein Res.，43：127-138，1994；Lu and Felix，Peptide Res.，6：142-6，1993；Felix et al.，Int.J.Peptide Res.，46：253-64，1995；Benhar et al.，J.Biol.Chem.，269：13398-404，1994；Brumeanu et al.，J Immunol.，154：3088-95，1995；也见Caliceti et al.(2003)Adv.Drug Deliv.Rev.55(10)：1261-77和Molineux(2003)Pharmacotherapy 23(8 Pt 2)：3S-8S)。聚乙二醇化也可用于在体内输送核酸分子。例如，腺病毒的聚乙二醇化可以增加稳定性和基因转移(见例如Cheng et al.(2003)Pharm.Res.20(9)：1444-51)。
希望的血液水平可以根据血浆水平确定通过持续输注活性物质而维持。应注意由于毒性或者骨髓、肝或肾的功能障碍，临床医生知道怎样及何时终止、中断或调整治疗至更低剂量。相反，如果临床反应不是足够的(除外毒性副作用)，临床医生也知道怎样及何时调整治疗至更高的水平。
药物组合物可以通过例如口服、肺部、胃肠外(肌内、腹膜内、静脉内(IV)或者皮下注射)、吸入(通过细粉制剂)、经皮、鼻、阴道、直肠、或者舌下途径给予，且可以配制为适于每种给予途径的剂量形式(见例如国际PCT申请No.WO 93/25221和WO 94/17784；欧洲专利申请613,683)。
选择的蛋白酶以对于所治疗的患者足以发挥治疗作用且不存在不希望的副作用的量包含于药物可接受的载体中。治疗有效浓度可以通过在已知的体外和体内系统中测试所述化合物而根据经验确定，如在本发明提供的测定方法中确定。
组合物中选择的蛋白酶的浓度依赖于复合物的吸收、失活和排泄速度、复合物的物理化学性质、剂量方案和给予量以及本领域技术人员已知的其它因素而定。用于治疗疾病或病症例如癌症或血管生成相关疾病的选择的蛋白酶的给予量可以通过标准临床技术确定。另外，可以应用体外测定法和动物模型以帮助鉴别最佳剂量范围。可以根据经验确定的精确剂量可以根据给予途径和疾病的严重性加以确定。
选择的蛋白酶可以一次给予，或者可以间隔一段时间以较小剂量分次给予。在整个治疗期间例如几个小时、几天、几周或几个月的期间内可以给予一或多次剂量的选择的蛋白酶。在一些情况中，可以持续给予。应理解精确的剂量和治疗持续时间是被治疗的疾病的函数，可以使用已知的测试方案根据经验而定或者通过体内或体外测试数据外推。注意浓度和剂量值也可以随着被减轻病症的严重性而变化。进一步地，对于任何特定的对象，特定的剂量方案应根据个体需要和管理或监督组合物给予的专业人士的判断随时调整，本文设定的浓度范围仅是举例说明，而不是限制所述组合物及含有其的组合的范围和应用。所述组合物可以每小时一次、每天一次、每周一次、每月一次、每年一次给予或者只给予一次。用于基因治疗中的含有所述多肽的组合物以及含有核酸的组合物的给予模式包括但不限于病变内(intralesional)、腹膜内、肌内和静脉内给予。本发明还包括输注、鞘内、皮下、脂质体介导的、depot介导的给予途径。本发明还包括经鼻、眼、口、表面、局部和耳给予途径。剂量可以根据经验确定，且依赖于适应证、给予模式和对象。举例的剂量包括0.1mg/天/kg对象体重、1mg/天/kg对象体重、10mg/天/kg对象体重、100mg/天/kg对象体重、200mg/天/kg对象体重和更多。
2.选择的蛋白酶的体内表达与基因治疗
选择的蛋白酶可以通过核酸分子的表达而输送至细胞和组织。选择的蛋白酶可以作为编码选择的蛋白酶的核酸分子通过离体技术和直接在体内表达而给予。
a.核酸的输送
核酸可以通过本领域技术人员已知的任何方法输送至细胞和组织。
i.载体-附加型载体和整合型载体
通过编码核酸分子的表达而给予选择的蛋白酶的方法包括重组载体的给予。所述载体可以设计为如通过包含复制起点而保留附加体或者可以设计为整合进细胞的染色体中。重组载体可包括病毒载体和非病毒载体。非限制性的病毒载体包括例如腺病毒载体、疱疹病毒载体、逆转录病毒载体，及本领域技术人员已知的任何其它病毒载体。非限制性非病毒载体包括人工染色体或者脂质体或者其它非病毒载体。选择的蛋白酶也可以用于使用病毒和非病毒载体的离体基因表达治疗中。例如，细胞可以工程化为表达选择的蛋白酶，例如通过将选择的蛋白酶编码核酸整合进基因组位置中，或者与调节序列可操纵地连接或者与调节序列可操纵地连接而置于基因组位置中。随后，这种细胞可以局部或者全身性地给予对象，例如给予需要治疗的患者。
选择的蛋白酶可以由病毒表达，给予需要治疗的对象。适于基因治疗的病毒载体包括腺病毒、腺伴随病毒、逆转录病毒及其它上述病毒。例如，腺病毒表达技术为本领域所熟知且腺病毒产生和给予方法也为本领域所熟知。腺病毒血清型可得自例如美国典型培养物保藏中心(ATCC，Rockville，MD)。腺病毒可以离体使用，例如从需要治疗的患者分离细胞并用选择的蛋白酶表达腺病毒载体转导细胞。在合适的培养期之后，经局部和/或全身性给予对象转导的细胞。或者，分离选择的蛋白酶表达腺病毒颗粒并且于药物可接受的载体中配制以输送治疗有效量来预防、治疗或改善对象的疾病或病症。典型地，腺病毒颗粒以1-1014个颗粒每kg对象体重的剂量输送，通常为106或108个颗粒至1012个颗粒每kg对象体重。在一些情况中，希望的是提供具有靶向细胞的物质的核酸来源，所述物质例如特异于细胞表面膜蛋白或者靶细胞的抗体或者靶细胞上受体的配体。
ii.人工染色体及其它非病毒载体输送方法
可以将核酸分子导入人工染色体及其它非病毒载体中。人工染色体(见例如美国专利No.6,077,697和国际PCT申请No.WO 02/097059)可以工程化为编码和表达所述同工型。
iii.脂质体和其它包囊化形式及含有核酸的细胞的给予
所述核酸可以包囊化在运载体如脂质体中或者导入细胞如细菌细胞、特别是减毒的细菌中或者导入病毒载体中。例如，当应用脂质体时，结合与胞吞作用相关的细胞表面膜蛋白的蛋白质可用于靶向和/或用于促进如下蛋白质的吸收：例如特定细胞类型向性的衣壳蛋白或其片段、在循环中经历内化的蛋白质的抗体，靶向胞内定位和增强胞内半衰期的蛋白质。
b.体外和离体输送
对于离体和体内方法而言，编码选择的蛋白酶的核酸分子被导入得自合适供体或治疗对象的细胞中。其中可以导入核酸以进行治疗的细胞包括例如适于治疗疾病或病症的任何希望的可利用的细胞类型，包括但不限于上皮细胞、内皮细胞、角质形成细胞、成纤维细胞、肌细胞、肝细胞；血细胞如T淋巴细胞、B淋巴细胞、单核细胞、巨噬细胞、嗜中性粒细胞、嗜酸性粒细胞、巨核细胞、粒细胞；各种干细胞或祖细胞，特别是造血干细胞或祖细胞，例如得自骨髓、脐带血、外周血、胎儿肝脏及其它来源的干细胞。
对于离体治疗，从与治疗对象相容的供体取出细胞或者从治疗对象中取出细胞，将核酸导入这些分离的细胞中并且将修饰的细胞给予所述对象。治疗包括直接给予，例如在多孔膜中包囊化，然后植入患者体内(见例如美国专利No.4,892,538和5,283,187)。适于在体外将核酸转移进入哺乳动物细胞中的技术包括使用脂质体和阳离子脂质(例如DOTMA、DOPE和DC-Chol)电穿孔、显微注射、细胞融合、DEAE-葡聚糖和磷酸钙沉淀方法。DNA输送方法可用于在体内表达选择的蛋白酶。这种方法包括核酸的脂质体输送方法和裸DNA输送，包括局部和全身性输送如使用电穿孔、超声和磷酸钙输送。其它技术包括显微注射、细胞融合、染色体介导的基因转移、微细胞介导的基因转移和原生质球融合。
选择的蛋白酶的体内表达可以与其它分子的表达相关联。例如，选择的蛋白酶的表达可以与细胞毒性产物的表达相关联，如在工程化的病毒中或者在细胞毒性病毒中。这种病毒可以靶向是治疗作用靶位的特定细胞类型。表达的选择的蛋白酶可用于增强病毒的细胞毒性。
选择的蛋白酶的体内表达可包括使选择的蛋白酶编码核酸分子与特异性调节序列可操纵地连接，所述调节序列如细胞特异性或者组织特异性启动子。选择的蛋白酶也可以从载体表达，所述载体在靶细胞类型和/或组织中特异性感染和/或复制。可诱导的启动子可用于选择性调节选择的蛋白酶表达。
c.全身性、局部和表面输送
核酸分子如裸核酸或者于载体、人工染色体、脂质体及其它运载体中的核酸分子可以通过全身性给予、表面、局部及其它给予途径给予对象。当全身性和体内给予时，核酸分子或者含有所述核酸分子的运载体可以靶向于细胞。
也可以直接给予，例如给予典型地靶向细胞或组织的载体或细胞。例如，肿瘤细胞和增殖性细胞可以是选择的蛋白酶体内表达的靶细胞。用于同工型的体内表达的细胞也包括患者自体细胞。这种细胞可以从患者体内取出，向其中导入表达选择的蛋白酶的核酸，然后通过注射或者移入给予患者。
2.组合治疗
本发明描述的任何选择的蛋白酶多肽和编码选择的蛋白酶多肽的核酸分子可以与其它治疗剂或者治疗方法组合应用，可以在其之前、交替或者随后应用，所述治疗剂或治疗方法包括但不限于其他生物制剂、小分子化合物和手术。对于任何疾病或病症、包括上文举例说明的所有那些疾病或病症，可以利用其它治疗剂或治疗方法，选择的蛋白酶多肽可以与其组合治疗这种疾病或病症。例如，本发明提供的用以治疗的增殖性疾病例如癌症的选择的蛋白酶多肽可以与其它抗癌治疗药物组合使用，在其之前、交替或者随后应用，所述其它抗癌治疗药物例如是化疗药物、放射性核素、放疗、细胞因子、生长因子、光敏剂、毒素、抗代谢物、信号调节剂、抗癌抗生素、抗癌抗体、血管生成抑制剂或其组合。在特殊实例中，本发明提供的用以治疗血栓形成疾病的选择的蛋白酶多肽可以与其它抗凝药物组合使用，在其之前、交替或者随后应用，所述其它抗凝药物包括但不限于血小板抑制剂、血管扩张剂、纤维蛋白溶解激活物或者其它抗凝剂。举例的抗凝剂包括肝素、香豆素、蛭素、阿司匹林、萘普生(naproxen)、甲氯芬那酸(meclofenamic acid)、布洛芬(ibuprofen)、吲哚美辛、phenylbutazare、噻氯匹定(ticlopidine)、链激酶、尿激酶和组织纤溶酶原激活物。
3.制造品与试剂盒
选择的蛋白酶多肽或者编码选择的蛋白酶多肽的核酸或者其衍生物或生物活性部分的药物化合物可以包装成制造品，其含有包装材料、有效治疗疾病或病症的药物组合物和标示用于治疗疾病或病症的选择的蛋白酶多肽或核酸分子的标记。
本发明提供的制造品含有包装材料。用于包装药物产品的包装材料为本领域技术人员所熟知。见例如美国专利No.5,323,907、5,052,558和5,033,252所述，所述文献均以其全部内容并入本文作参考。药物包装材料的例子包括但不限于泡包装(blister pack)、瓶、管、吸入器、泵、袋子、小瓶、容器、注射器和适于选择的制剂和指定给予和治疗模式的任何包装材料。本发明提供的化合物和组合物的大批制剂预期用于用于任何靶介导的疾病或病症的多种治疗。
选择的蛋白酶多肽和核酸也可以在试剂盒中提供。试剂盒可包括本文描述的药物组合物及给予项目(item)。例如，可以用给予装置如注射器、吸入器、剂量杯、滴管或者涂药器提供选择的蛋白酶。所述试剂盒可任选包括使用说明书，包括剂量、给药方案和给予模式说明书。试剂盒也可包括本发明描述的药物组合物和诊断项目(item)。例如，这种试剂盒可包括测量对象体内选择的蛋白酶的浓度、量或者活性的项目(item)。
J.举例的用选择的蛋白酶多肽的治疗方法
本发明提供的裂解特定靶的选择的蛋白酶多肽和编码本发明提供的选择的蛋白酶的核酸分子可用于治疗任何这样的疾病或病症，所述疾病或病症与含有靶序列的蛋白质相关或者对所述疾病或病症应用裂解所述靶序列的蛋白酶。例如，工程化为裂解tPA靶底物如纤溶酶原的选择的uPA多肽可用于治疗与tPA靶底物相关的或者对其应用tPA多肽的任何疾病或病症。与tPA靶底物相关的疾病的例子包括血栓溶解疾病(thrombolytic disorders)，其中用本发明提供的选择的蛋白酶治疗可以促进纤溶酶原裂解为其活性蛋白酶形式纤溶酶并且诱导血凝块的溶解。
选择的蛋白酶多肽单独具有治疗活性或者与其它药物组合具有治疗活性。本发明提供的选择的蛋白酶多肽被设计为呈现超过竞争结合蛋白的改良性质。这种性质包括例如可以改善所述多肽的治疗效率。本章节提供了举例性的应用和给予方法。这些描述的治疗方法只是举例而已，不限制选择的蛋白酶多肽的应用。
本发明提供的选择的蛋白酶多肽可以用于与含有靶序列的蛋白质相关的多种治疗方法以及诊断方法中。这些方法包括但不限于治疗下文描述和列举的生理学状况和病理学病症。本发明提供的选择的蛋白酶多肽与竞争性结合蛋白或者裂解特定靶的蛋白酶相比可以呈现出改良的体内活性和治疗作用，包括较低剂量即可达到相同作用、更持续的治疗作用和在给予和治疗方面的其它改善。使用选择的蛋白酶多肽在治疗方面得以改善的例子包括但不限于更好的靶组织渗透(例如肿瘤渗透)、更高的效力、更低的剂量、更低和/或更低频率的给予、降低的副作用和增加的治疗作用。特别是由于选择的蛋白酶可裂解并失活大量的靶底物，因此选择的蛋白酶提供实质扩大的治疗作用。
特别地，选择的蛋白酶多肽指定用于这样的治疗方法中，该治疗方法中已经使用裂解特定靶的蛋白酶进行治疗。这种方法包括但不限于治疗如下疾病和病症的方法，所述疾病或病症例如但不限于血液凝固功能障碍，包括血栓溶解疾病和弥漫性血管内凝血，心血管疾病，神经系统病症、增殖性疾病如癌症，炎症性疾病，自身免疫疾病，病毒感染，细菌感染，呼吸道疾病，胃肠道疾病和代谢疾病。
用选择的蛋白酶多肽治疗疾病和病症可以使用本文所述合适的制剂通过任何合适的给予途径得以实现，所述给予途径包括但不限于肌内、静脉内、皮内、腹膜内注射、皮下、硬膜外、鼻、口、直肠、局部、吸入、口腔(例如舌下)和经皮给予途径。如果需要，可以根据经验确定或者推断特定的剂量和持续时间及治疗方案。例如，裂解相似序列的野生型蛋白酶多肽的示例剂量可用作起始点以确定合适剂量。例如，重组tPA多肽的剂量可用作确定裂解tPA靶的选择的uPA多肽的剂量的指导。
剂量水平和方案可以基于已知剂量和方案确定，且如果需要则可以基于选择的蛋白酶多肽性质的改变加以推断和/或可以基于各种因素根据经验确定。如与其它相似蛋白酶相比选择的蛋白酶多肽的活性水平和半衰期这样的因素可用于进行这样的确定。特殊的剂量和方案可以根据经验加以确定。其它这样的因素包括个体体重、一般健康状况、年龄、所用特定化合物的活性、性别、饮食、给予时间、排泄速度、药物组合、疾病的严重性和进程和患者的疾病倾向及主治医生的判断。所述活性成分，选择的蛋白酶多肽，典型与药物有效载体组合。可以与所述载体材料组合以产生单剂量或多剂量形式的活性成分的量根据治疗的宿主和特定的给予模式而变化。
选择的蛋白酶多肽对于治疗疾病或者改善疾病症状的作用可以通过使用本领域已知的治疗特定疾病的任何诊断测试方法监测。在患者病症改善时，如果需要可以给予维持剂量的化合物或组合物；可以对剂量、剂量形式或者给予频率或者这些组合加以修改。在一些情况中，在疾病症状出现任何复发时或者根据预定剂量方案，对象可需要长期的间歇治疗。在其它情况中，可需要另外的给予以应答急性事件如出血、创伤或者手术过程。
在一些实例中，应用发挥蛋白酶激动剂(即模拟物)或者蛋白酶拮抗剂功能的所述选择的蛋白酶蛋白的变体。所述选择的蛋白酶多肽的变体可以通过诱变方法产生(例如分立的点突变或者蛋白酶蛋白的截短)。所述选择的蛋白酶多肽的激动剂可以基本上保留天然形式的选择的蛋白酶多肽的相同生物学活性或其亚类(subset)活性。所述选择的蛋白酶多肽的拮抗剂可以通过例如裂解与选择的蛋白酶多肽相同的靶蛋白从而抑制天然形式的选择的蛋白酶多肽的一或多种活性。因此，特异的生物学效应可以通过用受限功能的变体处理而激发。在一个实施方案中，用具有天然形式的选择的蛋白酶多肽的亚类生物学活性的变体处理对象，与用天然形式的选择的蛋白酶多肽处理相比在所述对象体内具有较少的副作用。
下文是选择的蛋白酶可以用作治疗剂单独或者与其它药物组合治疗的一些举例的疾病或病症。举例的蛋白酶选择的靶位只是举例说明而已，不限制用于本发明提供的方法中的可能靶位的范围。
1.裂解tPA靶位的选择的uPA多肽的举例的治疗方法
裂解tPA靶序列的选择的uPA多肽可用于改善血栓形成性疾病包括急性和慢性病症的治疗应用中。急性病症包括心脏病发作和中风，慢性病症包括动脉和深静脉血栓形成和再狭窄。选择的uPA多肽可以用作溶栓治疗剂用于改善这种病症的症状。治疗性组合物包括所述多肽、单独的cDNA分子或者部分病毒载体输送系统或者其它基于载体的基因表达输送系统，存在于脂质体输送系统中等。用作溶栓治疗剂的组合物通常是于药物合适的赋形剂中的生理学有效量的选择的uPA多肽。根据给予模式和治疗的病症，所述溶栓治疗剂以单剂量或多剂量给予。本领域技术人员意识到根据治疗的病症可以对剂量加以改变。
本发明提供的抑制或对抗血液凝固的选择的uPA多肽可用于治疗缺血性疾病的抗凝方法中，所述疾病如周围血管疾病、肺栓塞、静脉血栓形成、深静脉血栓形成(DVT)、浅表性血栓性静脉炎(SVT)、动脉血栓形成、心肌梗塞、短暂性脑缺血发作、不稳定型心绞痛、可逆性神经性缺乏(reversibleneurological deficit)、附属血栓溶解活性(adjunct thrombolytic activity)、过度凝血病症(excessive clotting conditions)、再灌注损伤、镰状细胞贫血或者中风。在过度凝血危险增加的患者中，如在DVT或SVT患者，手术期间，可以给予本发明提供的蛋白酶失活的选择的uPA多肽以防止手术中的过度凝血，所述手术例如但不限于是心脏手术、血管成形术、肺部手术、腹部手术、脊柱手术、脑部手术、血管手术或者器官移植手术，包括心、肺、胰腺或肝的移植。在一些情况中，单独用选择的uPA多肽进行治疗。在一些情况中，选择的uPA多肽与所治疗的病症或疾病所需的另外的抗凝因子联合给予。
tPA是食品和药品管理局(FDA)许可的用于急性血栓形成性中风的唯一疗法。tPA及其变体可以商购且已经许可给予人体治疗多种病症。例如，alteplase(Activase
，Genentech，South San Francisco，Calif.)是一种重组人tPA。Reteplase(Retavase
，Rapilysin
；Boehringer Mannheim，Roche Centoror)是一种重组的非糖基化形式的人tPA，其中所述分子已经被遗传工程化为含有原始蛋白质527个氨基酸中的355个氨基酸。Tenecteplase(TNKase
Genentech)是人tPA的527个氨基酸糖蛋白衍生物，其与天然人tPA的不同之处在于具有三个氨基酸取代。这些取代降低了血浆清除率、增加了纤维蛋白结合(并从而增加纤维蛋白特异性)且增加了对于纤溶酶原激活物抑制剂-1(PAI-1)的抗性。Anistreplase(Eminase
，SmithKline Beecham)是另一种可商购的人tPA。本发明提供的对于tPA靶具有特异性的选择的uPA多肽可以被相似地修饰并且用于可使用tPA治疗的任何治疗方法的处方中。
a.血栓形成性疾病和病症
血栓形成性疾病特征在于有利于血凝块形成的高凝性或者止血功能失调。举例的血栓形成性疾病和病症包括动脉血栓形成、静脉血栓形成、静脉血栓栓塞、肺栓塞、深静脉血栓形成、中风、缺血性中风、心肌梗塞、不稳定型心绞痛、心房纤维性颤动、肾损伤、经皮腔内冠状动脉成形术、弥漫性血管内凝血、败血症、人工器官、分流器(shunt)或假体(prostheses)，及其它获得性血栓形成性疾病，如下文讨论。治疗血栓形成性疾病的典型方法包括抗凝疗法，包括抑制凝血级联。
本发明提供的选择的uPA多肽和编码所述选择的uPA多肽的核酸可以用于针对血栓形成性疾病和病症的抗凝疗法中，包括治疗包含血管内凝血的病症。可以使用本发明提供的可抑制血液凝固的选择的uPA多肽用以例如控制血栓形成、溶解血栓或者防止血栓形成。在特殊的实施方案中，所述选择的uPA多肽和编码所述选择的uPA多肽的核酸可以用于治疗动脉血栓形成性疾病。在另一个实施方案中，所述选择的uPA多肽和编码修饰的选择的uPA多肽的核酸可以用于治疗静脉血栓形成性疾病，例如深静脉血栓形成。在一个特殊的实施方案中，所述选择的uPA多肽和编码选择的uPA多肽的核酸可用于治疗缺血性疾病如中风。使用选择的uPA多肽获得的治疗改善的例子包括例如但不限于更低的剂量、较少和/或更低频率的给药、降低的副作用及增加的治疗作用。可以例如通过使用动物模型检测选择的uPA多肽的治疗效力。例如，可以用选择的uPA多肽治疗缺血性中风小鼠模型或者任何其它已知的血栓形成性疾病或病症的疾病模型(Dodds，AnnNY Acad Sci 516：631-635(1987))。监测疾病症状和表型的进展以评估选择的uPA多肽的作用。选择的uPA多肽也可以给予动物模型以及在临床试验中给予对象，与安慰剂对照组相比评估在体内的效力。
i.动脉血栓形成
动脉血栓形成是动脉管壁破裂的结果。最通常的破裂发生在患有血管疾病的患者中如动脉粥样硬化患者。动脉血栓通常在被扰乱的血流区域以及在由于动脉粥样硬化斑所致的破裂部位形成，其将血栓形成内皮下膜(thrombogenic subendothelium)暴露于血小板和凝血蛋白，由此激活凝血级联反应。斑破裂也可以由于斑块内出血导致血管进一步狭窄。非闭塞性血栓可以与血管壁组合在一起，并且可以加速动脉粥样硬化斑的生长。形成的动脉血栓可以通过阻碍血流或者通过在远端微循环中栓塞而导致缺血。抑制血小板功能和凝血级联反应的抗凝剂和药物可以有效地防止和治疗动脉血栓形成。这类药物在治疗动脉血栓形成中是有效的。动脉血栓形成可以导致不稳定型心绞痛和急性心肌梗塞。本发明提供的抑制凝血的选择的uPA多肽可用于治疗和/或预防动脉血栓形成性疾病和病症，如不稳定型心绞痛和急性心肌梗塞。
ii.静脉血栓形成和血栓栓塞症
静脉血栓形成是其中由于血凝块形成与血凝块溶解的信号机制不平衡而在静脉中形成血凝块的病症，特别是在静脉系统血流缓慢的情况中。血栓形成的结果可包括对静脉和静脉瓣的损害，虽然静脉壁典型保持完整。血凝块通常可栓塞或者中断血流，且可以在血流中行进，进入器官如肺(肺栓塞)、脑(缺血性中风、短暂脑缺血发作)、心(心脏病发作/心肌梗塞，不稳定型心绞痛)、皮肤(暴发性紫癜)和肾上腺。在一些情况中，血流的阻断可导致死亡。具有复发静脉血栓栓塞倾向的患者的特征是具有血栓形成倾向。发生血栓栓塞性疾病的危险因素包括创伤、静止不动、恶性疾病、心力衰竭、肥胖、高雌激素水平、下肢瘫痪、心肌梗塞、静脉曲张、癌症、脱水、吸烟、口服避孕药和妊娠。对具有血栓形成倾向的家族进行遗传学研究表明可遗传的高水平凝血因子，包括FVIII、FIX和FXI(Lavigne et al.J.Thromb.Haemost.1：2134-2130(2003))。
深静脉血栓形成(DVT)是指下肢深静脉中静脉血凝块形成。导致DVT的三种主要因素是静脉内壁(vein lining)损伤、血液凝固倾向增加和血流缓慢。这些因素统称为Virchow’s三联征。静脉可以在在创伤或手术期间受损，或者由于疾病如Buerger’s病和DIC或者另外的血凝而受损。导致DVT发生的其它因素是与导致如上述更普通的血栓栓塞性疾病的因素相似的那些。在DVT中形成的血凝块仅导致轻度炎症，因此其更容易脱离而进入血流中。通常由于下肢肌肉的微弱收缩而可使血栓脱落。一旦血栓成为栓子，其可进入肺部血管，在此可导致肺梗死。在其血流中具有高水平活性FIX的患者发生深静脉血栓形成的危险增加(Weltermann et al.J.Thromb.Haemost.1(1)：16-18(2003))。
血栓栓塞性疾病可以遗传，其中所述疾病是由于凝血因子的遗传异常，因此导致止血机制不平衡而引起。一些先天缺陷包括抗凝血酶III、蛋白C、蛋白S或者纤溶酶原。其它因素包括对激活的蛋白C的抗性(也称作APC抗性或者因子V leiden效应，其中因子V中的突变使其对于蛋白C降解具有抗性)、前凝血酶中突变、异常纤维蛋白原血症(dysfibrinogemia)(赋予对纤维蛋白溶解的抗性)和过高半胱氨酸血症(hyperhomocysteinemia)。较年轻患者中的血栓栓塞性疾病的发生最常见是于上述先天缺陷，称作青少年血栓形成倾向(Juvenile Thrombophilia)。
对于静脉血栓栓塞性疾病和DVT的治疗典型包括抗凝治疗，其中给予口服剂量的肝素和新双香豆素。肝素通常输注给患者以控制急性事件，随后长期口服新双香豆素抗凝剂以控制进一步发作。其它治疗包括直接凝血酶抑制剂，血小板功能抑制剂如阿司匹林和葡聚糖以及对抗静脉淤滞的治疗包括应用加压袜子和气动压缩装置。本发明提供的抑制血液凝固的选择的uPA多肽可用于针对血栓栓塞性疾病和/或DVT的抗凝治疗中。在一些实施方案中，本发明提供的抑制血液凝固的选择的uPA多肽可用于对表现出具有血栓栓塞性疾病和/或DVT危险因素的患者中血栓栓塞性疾病和/或DVT的预防治疗。
(a)缺血性中风
当流入脑部的血流被中断时发生缺血性中风，其中脑部循环的突然降低导致神经功能的相应丧失。与特征在于大脑内出血的出血性中风相反，缺血性中风通常是由于血栓形成或者栓子导致。缺血性中风占所有中风的大约80％。除了发生如上述血栓栓塞的原因和危险因素之外，导致脑动脉剥离(dissection)的过程(例如创伤、胸主动脉剥离、动脉炎)也可导致血栓形成性中风。其他原因包括狭窄或阻塞动脉远端的灌注不足或者2个脑动脉区域之间易损越岭区(vulnerable watershed region)的灌注不足。缺血性中风的治疗包括预防和治疗所述病症的抗凝治疗。本发明提供的抑制凝血的选择的uPA多肽可用于治疗和/或预防或降低缺血性中风的危险。
iii.获得性凝血功能障碍
获得性凝血功能障碍是一些疾病或病症的结果，例如维生素K缺乏、肝脏疾病、弥漫性血管内凝血(DIC)或者循环抗凝剂的开发利用。血液凝固中的缺陷是由于所述病症或疾病导致的凝血因子继发缺陷的结果。例如，当肝脏处于疾病状态时，从肝脏中产生的凝血因子通常减少。随着凝血因子的合成降低，导致纤维蛋白溶解增加，并且血小板减少症(血小板缺陷)增加。肝脏产生的凝血因子的减少也可以是由于暴发型肝炎或者妊娠急性脂肪肝导致。这种病症消耗可利用的凝血因子而促进血管内凝血。本发明提供的选择的uPA多肽可用于治疗获得性凝血功能障碍以减轻凝血因子的缺陷。
(a)弥漫性血管内凝血(DIC)
弥漫性血管内凝血(DIC)是特征在于广泛和进行中的凝血机制激活的疾病。在DIC中，参与凝血的凝血酶激活与血凝块的纤溶酶降解之间失去平衡。血管或微血管纤维蛋白沉积的结果可损及供给不同器官的血供，可导致器官衰竭。在亚急性或慢性DIC中，患者出现超凝血表型，过量的凝血酶形成使血栓形成，并且可以存在静脉血栓形成的症状和迹象。与急性DIC相反，亚急性或慢性DIC通过减少过多血栓形成的方法进行治疗，包括肝素、抗凝血酶III和激活的蛋白C治疗。本发明提供的选择的uPA多肽和编码本发明提供的选择的uPA多肽的核酸可用于治疗亚急性或慢性DIC。在一个实施方案中，所述亚急性或慢性DIC多肽和编码所述选择的uPA多肽的核酸可以与其它抗凝疗法组合应用。可以例如使用动物模型检测选择的uPA多肽的治疗效力。监测疾病症状和表型的进展以评估所述选择的uPA多肽的作用。选择的uPA多肽也可以给予动物模型以及如临床试验中的对象与安慰剂对照组相比评估其在体内的效力。
(b)细菌感染和牙周炎
微生物如细菌的全身性感染通常与DIA相关。凝血途径的上调可以部分通过引起导致高水平细胞因子的炎症应答的微生物的细胞膜成分(脂多糖或内毒素)或者细菌外毒素(例如葡萄球菌α毒素)介导。所述细胞因子可影响凝血机制的诱导。
细菌病原体如Porphyrus gingivalis是导致成人牙周炎的熟知的致病因素。Porphyrus gingivalis细菌产生精氨酸特异性半胱氨酸蛋白酶，其发挥毒力因子的功能(Grenier et al.J.Clin.Microbiol.25：738-740(1987)，Smalley etal.Oral Microbiol.Immunol.4：178-181(1989)，Marsh，et al.FEMS Microbiol.59：181-185(1989)，和Potempa et al.J.Biol.Chem.273：21648-21657(1998)。Porphyrus gingivalis产生两种蛋白酶，称作50kDa和95kDa gingipains R(分别为RgpB和HRgpA)。所述蛋白酶可蛋白酶解凝血因子并因此将其激活。在细菌感染期间，gingipains R释放进入血流中可导致不受控制的凝血级联反应激活，从而导致蛋白酶过量产生且增加诱导弥漫性血管内凝血(DIC)的可能性。凝血酶浓度的大量增加可进一步导致牙周炎部位牙槽骨被破骨细胞吸收。
本发明提供的抑制血液凝固的选择的uPA多肽和编码选择的uPA多肽的核酸可用于治疗牙周炎。可以在牙周炎模型中检测选择的uPA多肽对于气道反应性的治疗效力。这种模型可在动物中获得，所述动物如非人灵长类动物、狗、小鼠大鼠、仓鼠和豚鼠(Weinberg and Bral，J.of Periodontology26(6)，335-340)。选择的uPA多肽也可以给予动物模型以及如临床试验中的对象，与安慰剂相比评估其在体内的效力。
b.其它tPA靶相关病症
本发明提供的选择的uPA多肽也可用于治疗已经用tPA进行治疗的神经系统病症。tPA被认为调节生理过程，包括组织重塑和可塑性，因为tPA具有水解胞外基质蛋白及其它底物的能力(Gravanis and Tsirska(2004)Glia49：177-183)。经历了如中风或损伤(例如由于事故或手术所致)的患者通常患有神经系统损害，可以用本发明提供的选择的uPA多肽治疗。本发明提供的选择的uPA多肽可用于治疗患有各种神经系统疾病和病症的对象，所述疾病和病症包括但不限于神经变性疾病如多发性硬化症、肌萎缩性侧索硬化症、亚急性硬化全脑炎、帕金森氏病、亨廷顿病、肌营养不良以及由于营养缺乏或毒素(例如神经毒素，药物滥用)引起的病症。此外，选择的uPA多肽可用于提供认知能力增强和/或治疗认知能力下降，例如“良性老年健忘症”、“年龄相关记忆力损害”、“年龄相关认知能力下降”等(Petersen et al.，J Immunological Meth.257：107-116(2001))，以及阿尔茨海默病。
可以使用各种神经系统损伤的动物模型检测选择的uPA多肽。可以使用的模型包括但不限于用于血栓栓塞性中风的啮齿动物、兔、猫、狗或者灵长类动物模型(Krueger and Busch，Invest.Radiol.37：600-8(2002)；Guptaand Briyal，Indian J Physiol.Pharmacol.48：379-94(2004))，脊髓损伤模型(Webb et al.，Vet.Rec.155：225-30(2004))等。所述方法和组合物也可以在人体内进行检测。许多不同的方法，包括标准化测试和计分系统，可用于评估动物和人体中的运动、感觉、行为和/或认知功能的恢复。可以使用任何合适的方法。在一个实例中，可以使用American Spinal Injury Associationscore，其已经成为在人体测量感觉功能恢复情况的主要仪器。见例如Martinez-Arizala，J Rehabil.Res.Dev.40：35-9(2003)，Thomas and Noga，JRehabil.Res.Dev.40：25-33(2003)，Kesslak and Keirstead，J Spinal Cord Med.26：323-8(2003)举例的各种计分系统和方法。用于人体的优选的剂量范围可以通过在组织培养系统和在动物模型中检测所述物质而确定，考虑所述物质的效力和任何观测到的毒性而确定。
c.诊断方法
本发明提供的选择的uPA多肽可用于诊断方法中，所述诊断方法包括但不限于检测具有改变的活性的纤维蛋白和纤维蛋白降解产物的诊断测定。所述测定因此可诊断血栓形成性病症。其它诊断性应用包括含有针对所述选择的uPA多肽的抗体的试剂盒，这些为本领域技术人员所熟知。
2.举例的裂解VEGF或VEGFR靶的选择的蛋白酶多肽的治疗方法
血管内皮生长因子(VEGF)是一种细胞因子，其结合特定的细胞表面受体(VEGFR)并通过其发出信号以调节血管发生，在血管发生过程中新血管从现有脉管产生。病理性血管发生是指与疾病相关的血管化增加，包括如实体肿瘤生长(McMahon，(2000)Oncologist.5 Suppl 1：3-10)、黄斑变性和糖尿病等事件。在癌症中，实体肿瘤需要不断增加的血液供应以生长和转移。缺氧或致癌突变增加了肿瘤及周围基质细胞中VEGF和VEGF-R mRNA的水平，导致现有血管延伸以及新血管网形成。在湿黄斑变性(wet maculardegeneration)中，在黄斑下面形成异常的血管生长。这些血管渗出血液和液体进入黄斑，损害感光细胞。在糖尿病中，眼部血液的缺少也可导致失明。VEGF刺激眼周围毛细血管生长导致不正确发挥功能的病态血管产生。
已经鉴别了VEGF的三个酪氨酸激酶家族受体(VEGF-R-1/Flt-1、VEGF-R-2/Flk-1/KDR、VEGF-R-3/Flt-4)。KDR(小鼠同系物是Flk-1)是只在内皮细胞上表达的VEGF的高亲和性受体，Kd为400-800pM(Waltenberger，(1994)J Biol Chem.269(43)：26988-95)。已经鉴别了VEGF与KDR结合是为病理性血管发生所需的关键的内皮细胞特异性信号途径(Kim，(1993)Nature.362(6423)：841-4；Millauer，(1994)Nature.367(6463)：576-9；Yoshiji，(1999)Hepatology.30(5)：1179-86)。在配体结合时受体的二聚体化导致细胞质结构域的自发磷酸化，以及结合配偶体的募集，使得信号传播遍及细胞质并进入细胞核中，改变细胞生长程序。用可溶的VEGF-R2治疗肿瘤可抑制肿瘤生长(Lin，(1998)Cell Growth Differ.9(1)：49-58)，且对磷酸化的化学抑制导致肿瘤细胞凋亡(Shaheen，(1999)Cancer Res.59(21)：5412-6)。
由血管内皮细胞生长因子(VEGF)及其受体的信号传导参与病理性血管发生以及癌症肿瘤维管结构的迅速发展。阻断这个信号传导途径的药物可阻止肿瘤血液供应的生长和维持，导致肿瘤系统性死亡。近来抗-VEGF抗体AVASTINTM在转移的结肠癌患者中的成功应用证实VEGF是癌症的抗血管发生治疗方法的靶位。尽管获得了这些令人鼓舞的结果，但是应用抗VEGF治疗中仍存在肿瘤的进展。抗体影响VEGF功能的机制以及抗体怎样阻止肿瘤生长的机制还未知。敲低(Knock down)实验示出阻断VEGF功能即可阻断血管发生。因此，抑制通过VEGFR-2的血管发生信号传导代表对于开发具有新靶向的工程化蛋白酶的一个未揭示的理想治疗领域。
在人和小鼠实体肿瘤的多个小鼠模型中靶向VEGF受体和Flk-1/KDR的治疗方法单独应用或者与细胞毒性治疗方法(Klement，(2000)J Clin Invest.105(8)：R15-24)组合应用抑制病理性血管发生并示出肿瘤大小减小(Prewett，(1999)Cancer Res.59(20)：5209-18；Fong，(1999)Neoplasia 1(1)：31-41.Erratum in：(1999)Neoplasia 1(2)：183)。使用小分子抑制剂和抗体进行的研究证实VEGF受体家族是有效的抗血管发生靶位，但是仍需要更有效的治疗方法。
VEGFR由具有七个免疫球蛋白(Ig)样结构域的胞外区、一个跨膜区和两个细胞质酪氨酸激酶结构域组成。前三个Ig样结构域已经示出调节配体结合，而结构域4至7具有在不存在配体条件下抑制正确二聚体化和信号传导的作用。作为工程化蛋白酶的选择性蛋白酶解的靶位，其具有如下预期靶特性：易于被蛋白酶解的氨基酸不稳定区、人、大鼠与小鼠物种之间的高度序列相同性、在裂解时信号传导下调以及蛋白酶解产生能非生产性结合配体的可溶受体。VEGF-R2的一些区域可用于特异性蛋白酶解，包括在跨膜区之前的柄区域以及Ig样结构域之间的非结构环(unstructured loop)。在一个实例中，本发明提供的丝氨酸样蛋白酶可以工程化为裂解其跨膜结构域与细胞因子或生长因子结合结构域(例如VEGFR)之间的特异性靶受体。发挥使蛋白质受体束缚(tether)在细胞表面或者环区功能的柄区域从而从多肽链中球状结构域中脱离。
a.血管发生、癌症及其它细胞周期依赖性疾病或病症
本发明提供的示例的选择的蛋白酶裂解VEGF或VEGFR，其可调节血管发生。在细胞表面分子是在肿瘤血管发生中传导信号的VEGFR时，所述裂解可阻止癌症传播。例如，细胞表面结构域从VEGFR分子中裂解可以失活其传递胞外信号、尤其是细胞增殖信号的能力。没有血管发生供给肿瘤，则癌细胞通常不能增殖。在一个实施方案中，本发明提供的选择的蛋白酶因此用于治疗癌症。同样，VEGFR的裂解可用于在其它病理中调节血管发生，如在黄斑变性、炎症和糖尿病中。在一个实施方案中，裂解细胞周期进展中涉及的靶VEGF或VEGFR蛋白失活了所述蛋白质使细胞周期前进的能力。细胞周期不再行进，则癌细胞不能增殖。因此，本发明提供的裂解VEGF或VEGFR的选择的蛋白酶用于治疗癌症及其它细胞周期依赖性病理。
本发明提供的选择的蛋白酶还可裂解致肿瘤性的可溶蛋白质。裂解VEGF多肽阻止通过VEGF受体的信号传导并且降低血管发生，因此减少其中血管发生起作用的疾病，如癌症、黄斑变性、炎症和糖尿病。进一步地，VEGF信号传导可以在某些细胞类型中调节细胞周期。因此，本发明提供的裂解VEGF的选择的蛋白酶可用于治疗癌症和其它细胞周期依赖性病理。
b.使用裂解VEGF或VEGFR的选择的蛋白酶的组合治疗方法
在一个实施方案中，治疗如癌症等病理包括例如与至少一种抗癌药物组合给予需要治疗的对象治疗有效量的蛋白酶，所述蛋白酶特异性裂解并失活VEGF/VEGFR-2复合物的信号传导。已经证实抗血管发生疗法已经成功用于治疗实体癌症和血液学恶性肿瘤(见例如Ribatti et al.(2003)JHematother Stem Cell Res.12(1)，11-22)。因此，本发明提供的组合物用于抗血管发生疗法中可以便于治疗血液学和实体组织恶性肿瘤。本发明提供的治疗组合物和方法可以单独施用或者与本领域技术人员已知的任何其它合适的抗癌疗法组合施用。例如，本发明提供的选择的蛋白酶可以与AVASTINTM组合应用，或者代替AVASTINTM用于给予其能提供治疗益处的任何疗法中。
在一个实施方案中，抗癌剂是至少一种化疗剂。在相关的实施方案中，与至少一种放射疗法组合给予所述蛋白酶。采用组合疗法将削弱血管发生信号并产生降低游离VEGF水平的可溶受体库。在特异的实施方案中，本发明提供的选择的蛋白酶具有体外特异性，其匹配受体的关键区域、Flk-1/KDR柄，越过6氨基酸区域。
本发明提供的选择的蛋白酶多肽可以于含有一种以上治疗剂的组合物中给予。所述治疗剂可以相同或不同，且可以是例如治疗性放射性核素、药物、激素拮抗剂、受体拮抗剂、酶或者由另一种物质激活的酶原、自分泌物质、细胞因子或者本领域技术人员已知的任何合适的抗癌药物。在一个实施方案中，与选择的蛋白酶多肽共同给予的抗癌药物是AVASTINTM。可用于本发明提供的方法中的其它治疗剂包括毒素、抗DNA、抗RNA、放射性标记的寡核苷酸如反义寡核苷酸、抗蛋白质以及抗染色质细胞毒性或者抗微生物剂。其它治疗剂为本领域技术人员所已知，本发明明确了这种其它治疗剂在本发明提供的方法中的应用。
抗肿瘤剂可以是多种化疗剂的一种，如烷化剂、抗代谢物、激素、抗生素、抗体、抗癌生物制剂、Gleevec、秋水仙碱、长春花生物碱、L-天冬酰胺酶、甲基苄肼、羟基脲、米托坦(mitotane)、亚硝基脲或者咪唑氨甲酰。合适的药物是促进微管蛋白去极化或者阻止肿瘤细胞增殖的那些药物。所述化疗剂包括但不限于由食品和药品管理局和U.S.Department of Healthand Human Services编制的Orange Book of Approved Drug Products WithTherapeutic Equivalence Evaluations中列出的那些抗癌药物。除了上述化疗剂之外，本发明提供的丝氨酸蛋白酶样蛋白酶也可以与放射疗法一起给予。本发明包含本领域已知的其它治疗方法。
治疗剂可以是化疗剂。化疗剂为本领域所已知，包括至少紫杉烷类、氮芥、氮丙啶衍生物、烷基磺酸盐/酯、亚硝基脲、三氮烯、叶酸类似物、嘧啶类似物、嘌呤类似物、长春花生物碱、抗生素、酶、铂配位复合物、取代的脲、甲基肼衍生物、肾上腺皮质抑制剂或者拮抗剂。更特别地，所述化疗剂可以是选自如下非限制性组的一或多种物质：类固醇、黄体酮、雌激素、抗雌激素或者雄激素。更特别地，所述化疗剂可以是阿扎立平(azaribine)、博来霉素、薯司他丁-1(bryostatin-1)、白消安(busulfan)、卡莫司汀(carmustine)、苯丁酸氮芥(chlorambucil)、顺铂(cisplatin)、CPT-11、环磷酰胺、阿糖胞苷、达卡巴嗪(dacarbazine)、更生霉素(dactinomycin)、柔红霉素、地塞米松、二乙基己烯雌酚、阿霉素、炔雌醇、依托泊甙(etoposide)、氟尿嘧啶、氟甲睾酮、吉西他滨(gemcitabine)、己酸羟孕酮(hydroxyprogesterone caproate)、羟基脲、L-天门冬酰胺酶、甲酰四氢叶酸(leucovorin)、洛莫司汀(lomustine)、氮芥(mechlorethamine)、醋酸甲羟孕酮(medroprogesterone acetate)、醋酸甲地孕酮(megestrol acetate)、美法仑(melphalan)，巯嘌呤(mercaptopurine)，甲氨蝶呤(methotrexate)、光神霉素、丝裂霉素、米托坦(mitotane)、丁酸苯酯(phenyl butyrate)、泼尼松(prednisone)、丙卡巴肼(procarbazine)、司莫司汀(semustine)、链佐星(streptozocin)、他莫昔芬(tamoxifen)、紫杉烷类、紫杉醇(taxol)、丙酸睾酮(testosterone propionate)、沙立度胺(thalidomide)、硫鸟嘌呤、噻替派(thiotepa)、尿嘧啶氮芥(uracil mustard)、长春碱(vinblastine)，或者长春新碱(vincristine)。也可以应用任意组合的化疗剂。可以在给予所述丝氨酸蛋白酶样突变蛋白多肽之前、期间或者之后给予所述化疗剂。
与本发明提供的选择的蛋白酶多肽组合应用或者共同给予的其它合适的治疗剂选自如下物质组成的一组：放射性同位素、硼附加物(boronaddend)、免疫调剂物、毒素、光敏剂或染料、癌症化疗药、抗病毒药、抗真菌药、抗菌药、抗原虫药和化学增敏剂(chemosensitizing agent)(见美国专利No.4,925,648和4,932,412)。合适的化疗剂在例如REMINGTON′SPHARMACEUTICAL SCIENCES，19th Ed.(Mack Publishing Co.1995)及在Goodman and Gilman′s THE PHARMACOLOGICAL BASIS OFTHERAPEUTICS(Goodman et al.，Eds.Macmillan Publishing Co.，New York，1980 and 2001 editions)中描述。其它合适的化疗剂如实验药物为本领域技术人员所已知。此外，合适的治疗性放射性同位素选自如下组成的一组：ax射线发射体、β射线发射体、γ射线发射体、俄歇电子(Auger electron)发射体、发射α-粒子的中子俘获剂和由于电子俘获衰变的放射性同位素。优选地，所述放射性同位素选自如下组成的一组：225Ac、198Au、32P、131I、131I、90Y、186Re、188Re、67Cu、177Lu、213Bi、10B和211At。
在一种以上的治疗剂与本发明提供的选择的蛋白酶组合应用的情况中，所述治疗剂可以是相同类别或类型，或者可以是不同类别或类型的。例如，所述治疗剂可包含不同的放射性核素或者药物与放射性核素。
在另一个实施方案中，由于其个体能量发射而在不同距离均有效的不同同位素用作第一种和第二种治疗剂与本发明提供的蛋白酶组合应用。这种物质可用于更有效地治疗肿瘤，以及用于治疗例如在正常临床环境中患有不同大小的多个肿瘤的患者。
极少的同位素可用于治疗极小的肿瘤沉积和单细胞。在这些情况中，药物或毒素可以是更有效的治疗药物，用以与本发明提供的蛋白酶共同给予。因此，在一些实施方案中，同位素与非同位素如药物、毒素和中子俘获剂组合应用，并且与本发明提供的蛋白酶共同给予。已知许多药物和毒素对细胞具有细胞毒性作用，可以与本发明提供的蛋白酶组合应用。它们可见于药物和毒素目录中，如Merck Index、Goodman和Gilman等，及上文引用的参考文献所述。
干扰细胞内蛋白质合成的药物也可以与蛋白酶组合用于本发明的治疗方法中，这种药物为本领域技术人员所已知，包括嘌呤霉素、环己酰亚胺以及核糖核酸酶。
本发明提供的治疗方法可以用于癌症治疗。本领域熟知放射性同位素、药物和毒素可以与抗体或者抗体片段缀合，所述抗体或抗体片段特异性结合由癌细胞产生或者与其相关联的标记，并且这种抗体缀合物可用于使放射性同位素、药物或者毒素靶向于肿瘤部位以增强其治疗效力且使得副作用最小化。这些物质和方法的例子在Wawrzynczak and Thorpe(inIntroduction to the Cellular and Molecular Biology of Cancer，L.M.Franks andN.M.Teich，eds，Chapter 18，pp.378-410，Oxford University Press.Oxford，1986)、Immunoconjugates：Antibody Conjugates in Radioimaging and Therapyof Cancer(C.W.Vogel，ed.，3-300，Oxford University Press，N.Y.，1987)、Dillman，R.O.(CRC Critical Reviews in Oncology/Hematology 1：357，CRCPress，Inc.，1984)、Pastan et al.(Cell 47：641，1986)、Vitetta et al.(Science238：1098-1104，1987)以及Brady et al.(Int.J.Rad.Oncol.Biol.Phys.13：1535-1544，1987)中综述。免疫缀合物在癌症及其它形式治疗中的应用的其它例子已经在美国专利No.4,331,647、4,348,376、4,361,544、4,468,457、4,444,744、4,460,459、4,460,561、4,624,846、4,818,709、4,046,722、4,671,958、4,046,784、5,332,567、5,443,953、5,541,297、5,601,825、5,635,603、5,637,288、5,677,427、5,686,578、5,698,178、5,789,554、5,922,302、6,187,287和6,319,500中揭示。
此外，本发明提供的治疗方法包括其中选择的蛋白酶与其它化合物或者技术组合应用以预防、减轻或者逆转某些细胞毒性物质的副作用。这种组合的例子包括例如一起给予IL-1和抗体以快速清除，如美国专利4,624,846所述。这种给予可以在给予最初的治疗处理之后3-72小时进行，所述最初的治疗处理是使用粒酶B突变蛋白或者MT-SP1突变蛋白组合抗癌药物(例如放射性同位素、药物或毒素作为细胞毒性成分)进行治疗。这可用于增强所述缀合物、药物或毒素从循环中清除以及用于减轻或逆转由所述治疗药物引起的骨髓和其它造血毒性。
在另一个实例中及如上文所述，癌症治疗可包括一种以上杀肿瘤剂的组合，例如药物与放射性同位素组合，或者放射性同位素与Boron-10药物组合进行被中子激活的治疗，或者药物与生物应答修饰剂组合，或者融合分子缀合物与生物应答修饰剂组合。细胞因子可以整合进这种治疗方案中以使得每种成分的效力最大化。
相似地，某些抗白血病和抗淋巴瘤抗体与是β或α射线发射体的放射性同位素组合，当这些物质不是只针对肿瘤细胞时可诱导骨髓及其它造血机能副作用。在肿瘤细胞处于循环和造血器官中时特别观测到这个结果。同时和/或随后给予至少一种造血细胞因子(例如生长因子，如集落刺激因子如G-CSF和GM-CSF)优选降低或改善所述造血副作用，同时增强抗癌作用。
本领域熟知许多放射性核素治疗方法可用于治疗癌症及其它病症，例如Harbert，″Nuclear Medicine Therapy″，New York，Thieme Medical Publishers，1087，pp.1-340所述。临床主治医生可以容易地调整所述细胞因子辅助疗法适合这种方法以减轻其任何造血副作用。相似地，可以共同给予细胞毒性药物与蛋白酶突变蛋白以例如治疗癌症、感染性疾病或者自身免疫疾病以及用于器官排斥反应治疗。这种治疗取决于与使用同位素或放射标记的抗体进行放射同位素治疗相似的原则。因此，临床医生能调整细胞因子的使用，通过在最初的抗癌治疗之前、期间和/或之后给予细胞因子从而减轻骨髓抑制及其它这种造血副作用。
3.举例的裂解补体蛋白靶的选择的MT-SP1多肽的治疗方法
本发明提供的蛋白酶多肽和核酸分子可以用于治疗其中包含补体途径激活的任何病症，特别是炎症性病症，包括急性炎症性病症如感染性休克，以及慢性炎症性病症如类风湿性关节炎(RA)。急性炎症性病症可以是免疫介导的疾病例如自身免疫系统疾病或者由免疫复合物介导的炎症引起的组织损伤。补体介导的炎症性病症也可以是具有炎症性成分的神经变性疾病或者心血管疾病。这个章节提供了蛋白酶的一些举例的应用和给予方法。这些描述的治疗只是举例而不是限制蛋白酶的应用。这些方法包括但不限于下文描述和列举的生理状况和病理病症的治疗方法。这些方法包括但不限于治疗败血症、类风湿性关节炎(RA)、膜增生性肾小球肾炎(MPGN)、红斑狼疮、多发性硬化(MS)、重症肌无力(MG)、哮喘、炎症性肠病、呼吸窘迫综合症、免疫复合物(IC)介导的急性炎症性组织损伤、多器官衰竭、阿尔茨海默病(AD)、缺血-再灌注损伤，或者Guillan-Barre综合征的方法，所述缺血-再灌注损伤是由于例如心肌梗塞(MI)、中风、心肺转流术(CPB)或者冠状动脉旁路移植术、血管成形术或血液透析等事件或治疗引起的。
用蛋白酶治疗疾病和病症可以通过使用如本文所述的合适制剂并通过任何合适的给予途径而实现，所述给予途径包括但不限于皮下注射、口服和经皮给予等。如果需要，可以根据经验确定或推断特定的剂量和持续时间以及治疗方案。例如，重组和天然蛋白酶多肽的示例剂量可以用作起始点以确定合适剂量。与野生型或支架蛋白酶相比具有更加特异性和/或选择性的修饰的蛋白酶在降低的剂量和/或频率下可以是有效的。剂量水平可以基于多种因素加以确定，例如个体体重、一般健康状况、年龄、应用的特定化合物的活性、性别、饮食、给药时间、排泄速度、药物组合、疾病的严重性和进程以及患者的疾病倾向和主治医生的判断。可以与载体材料组合产生单剂量形式的活性成分的量根据治疗的宿主和特定的给予模式而有所不同。
在患者的病症得以改善的情况下，如果需要，则可以给予维持剂量的化合物或组合物；可以对所述剂量、剂量形式或者给药频率或者这些组合进行修改。在这些情况中，在疾病症状出现任何复发的情况下需要给予对象长期间歇治疗。
a.免疫介导的炎症性疾病
本文所述方法中选择的蛋白酶及修饰的蛋白酶包括但不限于本发明提供的变体MT-SP1蛋白酶可用于治疗炎症性疾病。可以用蛋白酶治疗的炎症性疾病包括急性和慢性炎症性疾病。举例的炎症性疾病包括中枢神经系统疾病(CNS)、自身免疫系统疾病、气道超反应性病症如哮喘、类风湿性关节炎、炎症性肠病和免疫复合物(IC)介导的急性炎症性组织损伤。
实验性自身免疫性脑脊髓炎(EAE)可以作为多发性硬化症(MS)的模型(Piddlesden et al.，(1994)J Immunol 152：5477)。EAE可以在许多遗传易感物种中诱导，通过用髓磷脂和髓磷脂成分如髓磷脂碱性蛋白、蛋白脂质蛋白和髓磷脂少突胶质细胞糖蛋白(MOG)免疫接种进行诱导。例如，MOG-诱导的EAE概括了人MS的基本特征，包括慢性复发性临床疾病进程，炎症的病理组织学三联征、反应性神经胶质增生以及大量汇合的脱髓鞘斑形成。蛋白酶及修饰的蛋白酶可以在EAE动物模型中评估。蛋白酶例如通过每日腹膜内注射给予，监测疾病症状的进程和进展，与对照动物对比。也可以测量使疾病恶化的炎症性补体成分的水平，通过在溶血测定中测定血清补体活性以及通过测定补体成分如C1、C3和C9的沉积进行测量。
补体激活在如类风湿性关节炎(RA)等疾病中调节炎症(Wang et al.，(1995)PNAS 92：8955)。蛋白酶及修饰的蛋白酶包括本发明提供的变体MT-SP1多肽可用于治疗RA。例如，蛋白酶可以通过局部或全身性注射给予。可以每日或每周给予一剂蛋白酶。聚乙二醇化的蛋白酶可用于降低免疫原性。在一个实例中，可以在小鼠中诱导II型胶原诱导的关节炎(CIA)，作为在组织学上与RA相似的自身免疫性炎症性关节疾病的模型，其特征在于炎症性滑膜炎、血管翳形成以及软骨和骨的侵蚀。为了诱导CIA，可以在存在完全弗氏剂的条件下将牛II型胶原(B-CII)皮内注射至尾基底部。21天后，使用相同方案再次免疫接种小鼠。为了检验蛋白酶或修饰的蛋白酶包括MT-SP1多肽的作用，在最初用B-CII攻击3周后，可以每周经腹膜内给予两次蛋白酶或对照物，共3周。在最初免疫接种后7周，处死小鼠进行组织学分析。为了评估蛋白酶对于确定疾病的治疗作用，在一或多个肢体发生临床关节炎症状之后可以每天给予蛋白酶，共10天。通过使用测径器测量脚掌厚度可以监测最初受侵袭的关节的肿胀程度。在这两个模型中，抽取小鼠血清进行溶血测定以及测量激活的补体标记例如C5a和C5b-9。在另一个实例中，灵长类动物模型可用于RA治疗。在用蛋白酶多肽和对照物治疗的对象中监测柔软和肿胀关节反应以评估蛋白酶治疗作用。
蛋白酶或修饰的蛋白酶包括但不限于本发明提供的变体MT-SP1多肽可用于治疗免疫复合物(IC)-介导的急性炎症性组织损伤。IC-介导的损伤是由针对组织中IC沉积的局部炎症应答引起的。随后的炎症应答特征在于水肿、中性、出血以及最终的组织坏死。IC-介导的组织损伤可以在体内Arthus(RPA)反应中研究。简而言之，在RPA反应中，将过量的抗体(例如兔IgG抗鸡卵白蛋白)注射进动物如大鼠或豚鼠的皮肤中，所述动物预先已经静脉内输注了相应抗原(即鸡卵白蛋白)(Szalai et al.，(2000)J Immunol 164：463)。就在立即引发RPA反应之前，可以在通过右侧股静脉通过静脉内注射给予相应抗原同时给予蛋白酶或者推注对照物。或者，蛋白酶可以在RPA反应的最初一小时期间给予，在注射所述抗原之后以及在皮内注射所述抗体之前立即开始。蛋白酶对于产生补体依赖性IC介导的组织损伤的作用可以在RPA开始后各个时间点评估，通过收集血液以确定血清溶血活性，以及通过收集皮肤感染区域以量化损伤区域大小。
治疗性蛋白酶如包括本发明提供的变体MT-SP1多肽可用于治疗败血症和可以导致致命休克的严重败血症。补体介导的致命休克的模型可用于测试蛋白酶作为治疗剂的作用。在一个这样的实例中，可以用微量的脂多糖(LPS)致敏(primed)大鼠，随后给予针对补体的膜抑制剂的单克隆抗体(抗-Crry)(Mizuno M et al.，(2002)Int Arch Allergy Immunol 127：55)。可以在起始致命休克进程的任何时间给予蛋白酶或对照物，例如在LPA致敏之前、LPS致敏之后、或者在给予抗-Crry之后给予，并且可以评估大鼠从致命休克中得以挽救的情况。
b.神经变性疾病
补体激活使得阿尔茨海默病(AD)进展恶化，并且导致AD患者脑中神经突丧失。本文描述的蛋白酶及修饰的蛋白酶包括但不限于本发明提供的变体MT-SP1多肽可用于治疗AD。模拟AD的一些神经病理学和行为特征的小鼠模型可用于评估蛋白酶的治疗作用。转基因小鼠模型的例子包括在攻击启动子(aggressive promoter)控制下将具有产生疾病的突变的人淀粉状前体蛋白(APP)或者早老蛋白1(PS1)蛋白导入小鼠中。这些小鼠出现AD的特征，包括β淀粉状斑块以及营养不良的神经突增加。具有APP和PS1突变蛋白的双转基因小鼠出现更大量的纤维状β淀粉状斑块并且示出与所述斑块相关的激活的神经胶质和补体因子。蛋白酶可以通过例如每日经腹膜内或静脉内注射给予，监测疾病症状的进程与进展，与对照动物进行对比。
c.心血管疾病
本文所述的蛋白酶及修饰的蛋白酶包括但不限于本发明提供的变体MT-SP1蛋白酶可用于治疗心血管疾病。可使用蛋白酶治疗的心血管疾病包括由于中风、心肌梗塞、心肺转流术、冠状动脉旁路移植术、血管成形术或者血液透析所致缺血再灌注损伤。蛋白酶也可以用于治疗与心肺转流术相关的炎症反应，其可导致组织损伤。通常蛋白酶可以在诱导补体介导的缺血再灌注损伤的治疗或事件之前、同时或者随后给予。在一个实例中，蛋白酶可以在通过补体介导的缺血损伤诱导事件例如冠状动脉旁路移植术或者血管成形术治疗对象之前给予对象。
蛋白酶对于缺血性再灌注损伤的治疗作用可以在所述损伤的动物模型中评估。在一个这样的模型中，在大鼠中诱导心肌缺血，在所述大鼠前心包作一切口，围绕冠状动脉左前降支(LAD)距其起点5-8mm处放置3-0丝线，系紧，由此血管变得完全闭塞(Buerke et al.，(2001)J Immunol 167：5375)。经静脉内给予蛋白酶例如本发明提供的变体MT-SP1多肽或者对照运载体如盐水，随着冠状动脉闭塞后55分钟(即再灌注之前5分钟)推注增加剂量的蛋白酶或对照物，5分钟后(即缺血共60分钟后)，解开LAD处所系丝线，使缺血心肌再灌注3小时。在再灌注结束时，系紧围绕LAD的丝线。分析蛋白酶对于缺血性损伤的作用，可以通过评估对于心肌坏死、血浆肌酸激酶水平以及嗜中性粒细胞激活标记例如髓过氧化物酶活性和超氧化物自由基释放的作用加以评估。
在用含有6％人血浆的Krebs-Henseleit缓冲液灌注分离的小鼠心脏维持的补体介导的心肌损伤的另一个模型中，蛋白酶或修饰的蛋白酶处理可用于限制对心脏的组织损伤。在这个实例中，用于灌注心脏的缓冲液可以补加不同剂量的蛋白酶例如但不限于变体蛋白酶包括本发明提供的MT-SP1多肽。测定被灌注的心脏的人C3和C5b-9的沉积、冠状动脉灌注压、舒张期末压和心率。
蛋白酶及修饰的蛋白酶例如本发明提供的变体MT-SP1多肽在心肺转流术(CPB)或者冠状动脉旁路移植术之前或之后可以用作治疗剂以抑制在旁路转流术之后通常发生的且可引起组织损伤的炎症性免疫应答。全血在体外旁路转流中的体外再循环可用以刺激血小板和白细胞改变以及CPB诱导的补体激活(Rinder et al.(1995)J.Clin.Invest.96：1564)。在这个模型中，在血液马上进行体外循环之前可以将不同剂量的蛋白酶或修饰的蛋白酶或者对照缓冲液加入已经含有来自健康供体的血液和猪肝素的转移袋中。可以在再循环之后第5、15、30、45、60、75和90分钟取血样，进行补体研究测定例如溶血测定和/或补体激活测定以测量C5a、C3a和/或sC5b-9。在加入体外循环之前抽取的血液预处理样品可用作对照。可以对血样进行流式细胞术以确定血液循环中的白细胞(即嗜中性粒细胞)群上粘附分子的水平，例如CD11b和P-选择蛋白水平。
K.实施例
下述实施例仅用于举例目的，不意味着限制本发明的范围。
实施例1
突变体PAI-1抑制剂
A.突变体PAI-1抑制剂的表达和纯化
携带人PAI-1的cDNA(编码如SEQ ID NO：396所示的含有N末端Met的成熟PAI-1)的重组质粒pPAIST7HS被用作模板以将修饰导入到PAI-1反应中心环的氨基酸序列中。质粒pPAISTHS是质粒pPAIST7的衍生物，缺少在核苷酸对1的HindIII位点和在核苷酸对2106的SalI位点。质粒pPAIST7如(Franke et al.(1990)Biochimic et Biophysica Acta 1037：16-23)所述产生。简而言之，PAI-1 cDNA克隆pPAI-11RB用限制性核酸内切酶ApaLI和PflMI切割，含有2bp的PAI-1的残基1的密码子和379个残基蛋白质的残基2-376的全部编码序列的1127bp的PAI-1片段用凝胶电泳纯化。构建合成接头以重建PAI-1 cDNA编码序列的两个末端并在编码成熟PAI-1的第一个残基的三联体立即之前导入ATG蛋白质合成起始密码子以产生具有SEQ ID NO：396所示氨基酸序列的成熟PAI-1。另外，为了促进将该cDNA编码区插入质粒pBR322，设计接头以分别在PAI-1 cDNA片段的5’和3’末端产生EcoRI和HindIII限制性核酸内切酶位点。合成接头如下：N末端，5’AATTCTATGG-3’(SEQ ID NO：392)和5’-TGCACCATAG-3’(SEQ ID NO：393)；C-末端，5’-ATGGAACCCTGAA-3’(SEQ ID NO：394)和5’-AGCTTCAGGGTTCCATCAC-3’(SEQ ID NO：395)。接头在使用之前用多核苷酸激酶处理。合成接头(5’末端10bp，3’末端13bp)然后与用EcoRI和HindIII消化的1127bp ApaLI-PflMI DNA片段连接，通过凝胶电泳分离1146bp EcoRI-HindIII片段并克隆进EcoRI和HindIII裂解的pBR322中(SEQ ID NO：377)。
为起始构建pPAIST-7表达质粒，pBR322载体中的亚克隆用EcoRI裂解，线性质粒用细菌碱性磷酸酶去磷酸化。使用来自pC5A-48的含有trp启动子和核糖体结合位点的360bp EcoRI DNA片段，在标准连接后产生pPAIST-7。
为了产生pPAIST7HS原核表达载体(Shubeita et al.(1990)J Biol.Chem.，265：18379-18385)，pPAIST7用HindIII部分消化以线性化该质粒，用大肠杆菌DNA聚合酶I的Klenow片段平末端化并连接以消除上游HindIII位点。pPAIST7中PAI-1编码序列下游HindIII和SalI位点之间的序列的缺失及SalI位点的消除通过依次部分SalI消化、完全HindIII消化、用大肠杆菌DNA聚合酶I的Klenow片段的平末端化及连接而实现。
诱变反应用多位点诱变试剂盒(Stratagene)根据供应商限定的条件进行。产生野生型PAI-1中的相应于氨基酸VSARM(SEQ ID NO：378)的反应中心环的位置P4-P1’的氨基酸的诱变。制备了含有用RRARM(SEQ IDNO：379)序列置换PAI-1反应中心环从P4至P1’位置的野生型氨基酸序列VSARM的突变体PAI-1(PAI-1/RRAR)。RRARM诱变引物的序列为5’-CCACAGCTGTCATAAGGAGGGCCAGAATGGCCCCCGAGGAGATC-3’(SEQ ID NO：380)。制备了第二个突变体PAI-1(PAI-1/69)，其含有用PFGRS(SEQ ID NO：389)序列置换PAI-1反应中心环从P4至P1’位置的野生型氨基酸序列VSARM。PFGRS诱变引物的序列为：5’CCACAGCTGTCATACCCTTCGGCAGAAGCGCCCCCGAGGAGATC-3’(SEQ ID NO：390)。诱变之后，从转化体中分离的DNA经完全测序以证实希望的突变的存在及任何额外突变的不存在。
突变体PAI-1/RRAR和PAI-1/69用存在于pPAIST7HS载体中的N末端多组氨酸残基表达为融合蛋白。突变体PAI-1(即PAI-1/RRAR和PAI-1/69)的表达和纯化基于Ke et al.(J Biol.Chem.，272：16603-16609(1997))所述的方法。野生型和突变的PAI-1变体的表达通过将0.1μg编码突变体PAI-1的pPAIST7HS载体的DNA转化进大肠杆菌菌株BL21[DE3]pLyss(Novagen)中而实现，该菌株在异丙基-1-硫代-β-D-半乳吡喃糖苷存在下合成T7 RNA聚合酶。细菌培养物在37℃剧烈振荡下生长至吸光度A595为1.1-1.3，加入异丙基-1-硫代-β-D-半乳吡喃糖苷至终浓度为1mM以诱导T7RNA聚合酶的合成和PAI-1蛋白的产生。培养物在37℃再生长1-2小时，然后至30℃培养2-6小时。在4℃、8000×g离心20分钟沉淀细胞，并重悬于40ml冷的起始缓冲液(20mM乙酸钠，200mM NaCl，和0.01％Tween 20，pH 5.6)中。细胞悬浮液在32000×g超离心25分钟除去细胞碎片。
通过将含有可溶形式的PAI-1/RRAR或PAI-1/69的大肠杆菌裂解物注射到用CM-50Sephadex(Pharmacia，参见例如Sancho et al.(1994)Eur.J.Biochem.224，125-134)充填的XK-26柱(Pharmacia Biotech Inc)上纯化可溶的活性突变体PAI-1。用5柱体积的起始缓冲液(20mM乙酸钠，200mMNaCl，和0.01％Tween 20，pH 5.6)洗柱，用在相同缓冲液中的0.2-1.8M线性梯度NaCl洗脱PAI-1蛋白。收集峰级分，合并并用centriplus 30浓缩器(Amicon)浓缩。浓缩级分用于活性测量。
B.PAI-1活性测量
1.针对标准胰蛋白酶进行活性位点滴定
用Olson et al.(J.Biol.Chem.，270：30007(1995))所述的针对标准胰蛋白酶的活性位点滴定确定PAI-1/RRAR和PAI-1/69的活性浓度。简而言之，向1μM β-胰蛋白酶(Sigma)和10μM p-氨基苯甲脒探针(Sigma)的溶液中依次添加浓缩的抑制剂(0.5-6.0μM)。结合从荧光降低而监测，荧光降低伴随着当抑制剂结合时结合的探针从酶活性位点的置换。在添加每种浓缩的抑制剂后，进行1-2分钟的平衡，之后评估分别在325nm和345nm的激发和发射波长的荧光变化，以使结合的和游离的探针荧光之间的差异最大化。进行了仅探针与抑制剂在胰蛋白酶不存在下的对照滴定以校正背景荧光。抑制剂-酶滴定用线性回归分析拟合。
2.标准化t-PA制备物的滴定
突变体PAI-1还针对标准化t-PA制备物进行了滴定以评估活性。野生型PAI-1或突变体PAI-1的抑制活性经使用tPA(American Diagnostics，Inc，100U/μg)和生色tPA底物H-D-Ile-Pro-Arg-对硝基苯胺底物(S-2288，Chromogenix)的直接生色测定而测量。系列稀释的PAI-1(0.1-4.0μg)与t-PA在微滴定板中于室温保温一段固定时间(典型地20-60分钟)，通过加入生色底物S-2288至终浓度0.5mM而测量tPA的残余活性。与增加浓度的抑制剂保温后t-PA残余活性通过测量405nm的吸光度而评估。
实施例2
u-PA变体的噬菌体展示文库的构建
A.将野生型u-PA克隆进噬菌粒中
为证实u-PA的蛋白酶结构域在噬菌体上的功能展示，用引物496和497、Pfu DNA聚合酶及含有全长u-PA基因的cDNA(SEQ ID NO：474)作为模板的pCMV4质粒(SEQ ID NO：373)进行高可靠性PCR。PCR扩增中使用的引物如下：496，5’-ACGTGGCCCAGGCGGCCTTTCAGTGTGGCCAAAAG-3’(SEQ ID NO：374)；497，5’TCCTGGCCGGCCTGGCCGAGCAGGCCATTCTC-3’(SEQ ID NO：375)。两个引物均在5’末端携带SfiI酶限制位点(下划线)。纯化和SfiI消化后，PCR产物连接至SfiI消化的噬菌粒载体pComb3H(SEQ ID NO：376)(Andris-Widhopf et al.(2000)J ImmunolMethods，28：159-81)。该噬菌粒用作野生型或突变体u-PA的单价展示(参见下述)。这一构建体含有编码fd噬菌体的gIIIp基因的C-末端区的序列。在载体中存在具有两种具有不同识别序列的两个SfiI限制位点用于设计上述引物。用引物496和497扩增的PCR产物使得能将u-PA蛋白酶结构域cDNA(SEQ ID NO：475)定向克隆进噬菌粒(连接条件见下述)。在最终构建体中，含有u-PA蛋白酶结构域的序列的PCR产物克隆进OmpA和gIIIp序列的中间以将野生型或突变体u-PA展示为N-末端gIIIp融合体。
B.突变体u-PA的构建和突变体u-PA的噬菌体展示文库
为构建u-PA噬菌体展示文库，用含有uPA基因cDNA的pCMV4/u-PA质粒作为模板，如上所述在100μl反应混合物中使用由PCR Diversify诱变试剂盒(Clontech)提供的试剂进行25个循环的易错PCR扩增。采用合适的PCR条件以建立三种不同的PCR反应以使用来自以上的引物496和497通过根据厂商所述改变锰(MN2+)或dGTP的量而仅扩增uPA基因的蛋白酶结构域(SEQ ID NO：475)以实现掺入cDNA中的0.2、0.5或0.9％的突变率。扩增的PCR产物(805bp，SEQ ID NO：475)用PCR纯化试剂盒(Qiagen)纯化，随后以SfiI酶消化。来自每个诱变反应的SfiI消化的PCR产物用于产生三种不同的文库。
为了构建文库，pComb3H载体(SEQ ID NO：376)用SfiI酶消化，来自这一反应的较大DNA片段用“Gel slice kit”(Qiagen)凝胶纯化。进行三个独立的连接反应以用含有0.2、0.5和0.9％突变率的PCR产物构建3个文库。至少1μg凝胶纯化的载体与SfiI消化的PCR产物混合(1∶2比率)并将连接混合物在18℃保温过夜。接下来，连接混合物用Qiagen mini elute kit(Qiagen)纯化，并将DNA最终洗脱进60μl Milli Q纯化水中。纯化的DNA用基因脉冲仪(Bio Rad)电穿孔进400μl大肠杆菌XL Blue1电穿孔感受态细胞(Stratagene)中。细胞然后转移进10ml SOC培养基(Invitrogen)中并在摇床中在37℃保温1小时，之后在补加羧苄青霉素(75μg/mL)的大LB琼脂平板(245mm X 245mm)上铺板。在30℃过夜保温平板后，用细胞刮刀刮下转化体，所得培养物在补加羧苄青霉素(75μg/ml)的2X YT培养基中于37℃振荡培养2小时。培养物用辅助噬菌体(VCS M13)以MOI(感染复数)为5感染以用于扩增文库。在37℃振荡生长1小时后，培养物补加卡那霉素至终浓度为3μg/mL并在30℃振荡生长过夜。收获细胞，存在于上清中的噬菌体颗粒用PEG-NaCl溶液沉淀。同时，为了计算每个文库的多样性，等份电穿孔的细胞在补加羧苄青霉素(100μg/ml)的LB琼脂平板上铺板，在37℃保温过夜后计数菌落。使用相同方法产生后继代次的u-PA噬菌体展示文库以用于进一步改良针对PAI-1/RRAR抑制剂的鉴别的u-PA变体。从前面文库中鉴别的u-PA变体的DNA用作构建下一代文库的模板。
野生型或用随机诱变产生的u-PA噬菌体文库的催化效率用间接纤溶酶原激活测定分析。简而言之，5μl u-PA噬菌体(典型地～5×1012cfu)，0.2μMLys-纤溶酶原(American Diagnostic)和0.62mM Spectrozyme PL(AmericanDiagnostica)以总体积100μl存在。在微滴板中进行测定，405nm的光密度在Molecular Devices Thermomax中每30秒读取一次，共1小时。反应在37℃进行。还测定了u-PA或u-PA变体噬菌体的抑制。简而言之，5μl u-PA噬菌体(典型～5×1012cfu)与wt-PAI-1(0.1μM)或突变体PAI-1/RRAR抑制剂(1.0μM)混合并在室温保温30分钟，随后加入0.62mM Spectrozyme PL，0.2μM Lys-纤溶酶原(American Diagnostics，Inc.)。如上所述读取测定。
实施例3
从u-PA噬菌体文库中针对突变体抑制剂选择变体u-PA
A.u-PA噬菌体/突变体PAI-1复合物的选择
突变体PAI-1/RRAR抑制剂或PAI-1/69在淘选实验中用作“诱饵底物”以从实施例2描述的大的组合u-PA噬菌体展示文库中分离改变的u-PA噬菌体，所述改变的u-PA噬菌体对于突变体底物序列具有改良的反应性。简而言之，将含有来自所有三个u-PA噬菌体文库(0.2、0.5和0.9％诱变频率)的等表现度u-PA噬菌体的5μl(～2×1012至～1X1013)u-PA噬菌体与5μl诱饵底物(从0.1至1.0μM PAI-1/RRAR或者PAI-1/69)和10μl 10X间接缓冲液pH 7.4(0.5M Tris，1.0M NaCl，10mM EDTA，0.1％Tween 80)混合，总体积为100μl(即反应含有80μl H2O)。反应在室温下保温不同时间(典型为1小时，然而可以调节保温时间以控制溶液的严格性)。
使用CuSO4激活的琼脂糖捕获u-PA噬菌体-PAI-1抑制剂复合物。通过用CuSO4(100mM)处理完成对螯合琼脂糖(200μl；Pharmacia)的预激活。将CuSO4激活的琼脂糖用PBS缓冲液中的0.5％BSA在室温封闭1小时。将100μl的BSA封闭的琼脂糖珠在存在800μl的结合缓冲液(0.5M NaCl，20mM Tris，20mM咪唑，pH7.4)下加入100μl的上述淘选混合物反应中以捕获His-标记的PAI-1抑制剂-u-PA噬菌体复合物。在室温连续再保温1小时。接着，淘选混合物在2000rpm离心1分钟，含有结合的u-PA噬菌体-PAI-1抑制剂复合物的琼脂糖珠用1ml的结合缓冲液洗涤以除去未结合的噬菌体。洗涤步骤重复5-10次，含有结合的复合物的珠在每次洗涤后转移至新管中以避免任何非特异噬菌体的潜在“携带污染”。
结合的u-PA噬菌体-PAI-1/RRAR复合物在100μl洗脱缓冲液(0.5MEDTA)中洗脱。95μl等分的洗脱的噬菌体用于感染1ml XL-1Blue大肠杆菌细胞(0.6OD)以计算文库的输出(output)(表示在选择后获得的噬菌体数目)。感染的细菌铺板于含有羧苄青霉素(75μg/ml)的大平板上(245mm×245mm)以产生进行下一轮选择的文库。最后，其余的输出文库(即选择的噬菌体)用于制备各个噬菌体克隆以筛选和/或产生进行下一轮选择的新文库。根据希望的严格性水平，调节用于选择中的诱饵底物的浓度和所述文库与诱饵底物的保温时间。例如，可以选择这样的条件，由此所述文库的＜1％、2％、5％、10％或者10％以上的u-PA活性被含有诱饵底物序列的“诱饵PAI-1”抑制。典型地，使用较高浓度(例如0.5μM)的诱饵PAI-1进行第一轮选择，在后续轮的选择中诱饵丝氨酸蛋白酶抑制蛋白的浓度和与文库的保温时间降低。另外，在每轮选择时可以并行使用多重严格性。基于输出的质量(例如在成对的+/-诱饵丝氨酸蛋白酶抑制蛋白选择中噬菌体输出的信号与噪声比(见下文描述))和基于功能分析获得的“采样数(hit)”的质量，可以将来自一或多次这些选择的噬菌体输出用于接下来的选择中。
使用上述条件并行进行对照实验以将不用诱饵从u-PA文库中选择的噬菌体与从这个对照实验中选择的噬菌体与在存在诱饵丝氨酸蛋白酶抑制蛋白底物的条件下所述文库选择的输出进行比较。在存在诱饵的条件下选择的文库输出的cfu的范围通常比得自对照选择的输出高104至105。如果在对照选择中观测到较高的背景，则使用更严格的条件重复进行淘选，所述更严格的条件例如是减少保温时间、增加反应物(文库、诱饵或珠)浓度和增加洗涤与螯合琼脂糖结合的选择的噬菌体的数目和时间(直至10或更多倍)。
B.筛选对于新底物序列具有增加的反应性和催化效率的u-PA噬菌体
将等分的洗脱的u-PA噬菌体(5μl)与XL-1Blue大肠杆菌细胞(100μl)混合以感染，在37℃保温1小时。然后将感染的大肠杆菌细胞铺板于补加了羧苄青霉素(100μg/ml)的LB琼脂平板上。在37℃保温过夜后，挑取单独的菌落制备噬菌体。细胞在补加了羧苄青霉素(100μg/ml)和四环素(10μg/ml)的2ml的2×YT中生长，噬菌体制备如(Sambrook，J et al(1989)MolecularCloning，A laboratory manual，Cold spring Harbor laboratory)所述进行。在如下测定中检测鉴别的噬菌体以评定活性。将单独的噬菌体制品用于间接纤溶酶原激活测定中以鉴别活性噬菌体且将活性噬菌体制品用于抑制测定。
1.间接纤溶酶原激活测定
将单独的噬菌体制品用于间接纤溶酶原激活测定中以鉴别活性u-PA噬菌体。简而言之，在总体积100μl中存在5μl的u-PA噬菌体(典型为～5×1012cfu)、0.2μM Lys-纤溶酶原(American Diagnostic)和0.62mMSpectrozyme PL(American Diagnostica)。在微滴定平板中进行测定，在Molecular Devices Thermomax中每30秒读取在405nm的光密度，进行1小时。反应在37℃进行。
2.突变体PAI-1/RRAR对u-PA噬菌体的抑制
进一步检测突变体PAI-1/RRAR对于在间接纤溶酶原活性测定中鉴别的噬菌体的抑制作用。简而言之，对于抑制作用测定，将5μl活性u-PA噬菌体(典型为～5×1012cfu)一式两份加入微滴定平板的孔中，随后向一个孔中加入固定浓度的突变体PAI-1(例如1.0μM)，向另一个孔中加入磷酸盐缓冲盐水(PBS)。混合后，使反应在室温进行规定时间(例如30分钟)，随后加入0.62mM Spectrozyme PL、0.2μM Lys-纤溶酶原(American Diagnostics，Inc.)。对于对照实验，在相同条件下评定野生型u-PA噬菌体。在分光光度计中在405nm读取平板2小时。选择与野生型u-PA噬菌体相比对于PAI-1/RRAR或PAI-1/69呈现改良的敏感性的选择的u-PA噬菌体进行进一步分析和DNA测序。
3.肽底物筛选
另外，为了鉴别具有改良的催化效率的变体u-PA噬菌体，针对Ac-RRAR-AMC底物筛选各个噬菌体克隆。对于这个测定，将固定体积的噬菌体上清(例如35μl)与在1×间接测定缓冲液(50mM Tris，100mM NaCl，1mM EDTA，0.01％Tween 80)中的75μM Ac-RRAR-AMC底物混合，总体积为100μl。所述测定在96-孔或者384-孔黑色测定平板(Corning)中进行，使用分光光度计(Molecular Devices)读数器在380-450nm读取2小时。
为了证实改良，使用上述测定方法对在抑制剂和肽底物筛选之后鉴别的阳性u-PA噬菌体再次进行筛选。
C.选择的u-PA突变体的鉴别和鉴别的突变体的优化
将阳性噬菌体克隆与上述用于感染的XL-1Blue大肠杆菌细胞混合并将培养物在37℃振荡生长过夜。用质粒制备试剂盒(Qiagen)从所述过夜培养物纯化质粒DNA。将所述DNA送出进行商业测序，测序使用下述引物：535-5’-CAGCTATCGCGATTGCAG-3’(SEQ ID NO：381)；5542-5’GTGCGCAGCCATCCCGG-3’(SEQ ID NO：382)。突变体u-PA基因中改变的氨基酸残基在分析测试数据后被鉴别。
下表10示出从u-PA噬菌体文库针对PAI-1/RRAR突变体抑制剂进行的变体u-PA选择而鉴别的u-PA变体。下表10中示出的突变是采用胰凝乳蛋白酶编号。括号中的数字表示在噬菌体选择方法中突变体被鉴别的次数。基于活性测定的结果，u-PA噬菌体的最佳变体以下划线高亮示出。含有指定的突变的成熟u-PA前蛋白原(SEQ ID NO：433)的氨基酸序列如SEQ IDNO：434-445任一所示。
表10
| u-PA噬菌体文库变体PAI-1/RRAR | 突变体名称 | 突变位点(胰凝乳蛋白酶编号) | 修饰的氨基酸 | SEQID NO： |
| 选择I | AR73(2)AR81 | 3073，89 | Phe-IleuLeu-Ala，Ileu-Val | 434435 |
| 选择II&III | AR1AR3AR4(3) | 73217155 | Leu-ProArg-CysLeu-Pro | 436437438 |
| AR7AR32AR36AR37AR66AR24AR85 | 75，89，13813772，15513316038132，160 | Ser-Pro，Ileu-Val，Ileu-ThrGlu-GlyArg-Gly，Leu-ProGly-AspVal-AlaVal-AspPhe-Leu，Val-Ala | 439440441442443444445 |
在针对PAI-1/69抑制剂选择后展示对PAI-1/69抑制剂的增加的敏感性的突变体u-PA基因中改变的氨基酸残基在分析如上所述测序数据后被鉴别。从第一代蛋白酶噬菌体展示文库(I)中鉴别的突变体如下表11中所示。用实施例2B中所说方法使用PCR Diversify诱变试剂盒及引物496和497产生后续代次的蛋白酶噬菌体展示文库。对于第II代噬菌体展示文库，u-PA突变体u-PA/Ic用作模板进行诱变反应，该突变体含有相应于基于胰凝乳蛋白酶编号的F30V的突变。从第二代蛋白酶噬菌体展示文库(II)鉴别的突变体见下表11所示。对于第III代噬菌体展示文库，使用u-PA突变体u-PA-IIb或含有相应于基于胰凝乳蛋白酶编号的F30V/Y61(A)H或F30V/K82E的突变的u-PA-IIb突变体作为模板用于诱变反应。从第三代蛋白酶噬菌体展示文库(III)鉴别的突变体见下表11。对于第IV代噬菌体展示文库，u-PA突变体u-PA/IIIa用作模板用于诱变反应，所述突变体含有相应于基于胰凝乳蛋白酶编号的F30V/K82E/V159A的突变。从第四代蛋白酶噬菌体展示文库(IV)鉴别的突变体见下表11。括号中的数字代表具有相同突变的u-PA噬菌体被选择的次数。下划线表示由突变体获得的新突变。含有指定突变的成熟u-PA前蛋白原(SEQ ID NO：433)的氨基酸序列如SEQ ID NO：460-472任一所示。
表11
| 噬菌体文库 | 突变体名称 | 突变位点 | 修饰的氨基酸 | SEQ IDNO： |
| I | u-PA/Iau-PA/Ibu-PA/Ic(2)u-PA/Id | 21243030 | Phe-ValIle-LeuPhe-ValPhe-Leu | 460461462463 |
| II | u-PA/IIau-PA/IIb | 30，61(A)30，82 | Tyr-HisLys-Glu | 464465 |
| u-PA/IIc | 30，156 | Lys-Thr | 466 | |
| III | u-PA/IIIa(8)u-PA/IIIbu-PA/IIIcu-PA/IIId(2) | 30，82，15930，82，39，15930，82，158，15930，61A，92 | Val-AlaThr-Ala，Val-AlaThr-Ala，Val-AlaLys-Glu | 467468469470 |
| IV | u-PA/IVau-PA/IVb | 30，82，159，80，89，18730，82，159，80，84，89，187 | Glu-Gly，Ile-Val，Lys-GluGlu-Gly，Glu-Lys，Ile-Val，Lys-Glu | 471472 |
1.在针对PAI-1/RRAR抑制剂的聚焦噬菌体展示文库中氨基酸残基30和155的优化和重组
为富集u-PA变体对PAI-1/RRAR抑制剂的敏感性，基于胰凝乳蛋白酶编号的氨基酸30和氨基酸155(在上表10所示的第一次选择中鉴别为热点)被靶向用于分别使用下述引物进行随机化和重组：
TC30-5’GCCCTGGNNSGCGGCCATC-3’(SEQ ID NO：383)
TC155-5’GGAGCAGNNSAAAATGACTG-3’(SEQ ID NO：384)
用Quick Change多位点定向诱变试剂盒(Stratagene)根据厂商所述条件进行诱变。简而言之，在用T4多核苷酸激酶(New England Biolabs)根据厂商所述条件磷酸化引物之后，进行三个不同反应用于残基30和155的随机化：1)30单独；2)155单独；及30加155一起。使用DNA构建体pComb3H/u-PA变体(pARF 81)(其与相应的野生型u-PA蛋白酶结构域序列相比携带在残基L73A和I89V的突变)用作诱变反应中的模板，反应使用引物TC30(SEQ ID NO：383)和TC155(SEQ ID NO：384)分别用于位置30和155的随机化。在另一反应中，这两个引物用于一起随机化pARF 81变体DNA中的位置30和155。使用多位点诱变试剂盒(Stratagene)根据供应商规定的条件进行诱变反应。诱变后，将反应产物转化进XL-1Blue大肠杆菌细胞进行文库构建(见上述实施例2)。扩增的u-PA噬菌体文库被用于选择上述实施例3A和3B所述的改良的u-PA变体。
下表12描述了针对来自聚焦u-PA噬菌体文库的变体u-PA的PAI-1/RRAR突变体抑制剂进行选择而鉴别的u-PA变体，其中所有变体均具有基于胰凝乳蛋白酶编号的在氨基酸残基L73A和I89V的背景突变。下表12所示的突变使用胰凝乳蛋白酶编号。括号中的数字代表在噬菌体选择方法中突变体被鉴别的次数。基于活性测定的结果，u-PA噬菌体的最佳变体被下划线高亮示出。含有指定突变的成熟u-PA前蛋白原(SEQ ID NO：433)的氨基酸序列如SEQ ID NO：446-459任一所示。
表12
| 聚焦u-PA文库变体PAI-1/RRAR | 突变体名称 | 30(Phe) | 155(Leu) | SEQ ID NO |
| 选择I&II | ARF2ARF6ARF11ARF17ARF16ARF33ARF35ARF36ARF37ARF43ARF47ARF48ARF103ARF115 | ThrLeuValGlyLeu-Met-MetLeuValLeuLeuGly | -----Val-Met---Met-Met | 446447448449450451452453454455456457458459 |
实施例4
通过COS细胞的瞬时转染表达修饰的u-PA酶
为在哺乳动物表达系统中表达变体u-PA酶，从噬菌体展示结果鉴别的阳性克隆被用作模板(携带突变体u-PA蛋白酶结构域序列的pComb3H)用于编码选择的u-PA变体基因的cDNA的PCR扩增。用下述引物进行重叠延伸PCR(Ho，S et al(1989)Gene 77，51-59)。
717-5’-TTTCAGTGTGGCCAAAAG-3’(SEQ ID NO：385)；
718，5’-CAGAGTCTTTTGGCCACA-3’(SEQ ID NO：386)；
850，5’-GGGGTACCGCCACCATGAGAGCCCTGCTGGCGCGC-3’(SEQID NO：387)；
851，5’-GCTCTAGATCATCAGAGGGCCAGGCCATTCTCT-3’(SEQ IDNO：388)。引物850和851分别携带KpnI和XbaI限制酶序列(下划线)。
如下所述，PCR以两步进行以实现突变的cDNAu-PA基因的全长扩增。在第一步中，PCR在100μl反应中进行，使用Pfu DNA聚合酶、引物850、718和作为模板的含有全长uPA基因(SEQ ID NO：474)的pCMV4扩增500bp产物(相应于u-PA的EGF和Kringle结构域)。类似地，进行另一PCR以使用引物851和717用合适的突变体u-PA-pComb3H作为模板扩增突变体u-PA蛋白酶结构域(800bp，即相应于与SEQ ID NO：475所示的野生型序列相对比的突变体序列)。将这两个PCR产物凝胶纯化并用于下一轮PCR扩增。在第二步中，凝胶纯化的PCR产物(各5μl)在含有引物850和851的100μl反应混合物中用作模板。引物717和718具有重叠的互补序列，使得在第二步PCR中扩增出全长u-PA cDNA(1.3kb)。所述PCR产物用PCR纯化试剂盒(Qiagen)纯化，然后用KpnI和XbaI限制酶消化。在用QIAquick柱(Qiagen)纯化后，全长u-PA基因与已预先用KpnI和XbaI消化的pCMV4哺乳动物表达载体(SEQ ID NO：373)连接。连接混合物电穿孔进大肠杆菌XL-1Blue细胞并在补加羧苄青霉素(100μg/ml)的LB平板上铺板。在37℃过夜保温平板后，挑取各个菌落并在2ml LB培养基中生长以用于质粒纯化。所述质粒用于使用下述测序引物对完整u-PA基因进行测序。UPAF1-5’ATGAGAGCCCTGCTGGCGCGCC-3’(SEQ ID NO：476)和UPAF2-5’GGAAAAGAGAATTCTACCG-3’(SEQ ID NO：477)。
用Midi质粒制备试剂盒(Qiagen)大量制备具有正确突变且无任何另外突变的突变体u-PA并用Bio-Rad Gene Pulser电穿孔进COS-1细胞中。将20μg cDNA，100μg载体DNA和大约107个COS-1细胞置于0.4-cm小池中，以320 V，960微法拉第和Ω＝∞进行电穿孔(Tachias et al.(1995)J Biol.Chem.，270：18319-18322)。在电穿孔后，转染的细胞在含有10％胎牛血清和5mM丁酸钠的DMEM培养基(Irvine Scientific)中在37℃过夜保温。然后用无血清培养基洗细胞并在DMEM中于37℃保温48小时。在与无血清培养基保温后，收集条件培养基并用于进一步鉴定。
实施例5
纯化的突变体u-PA的鉴定
A.酶浓度的测量
在条件培养基中单链形式的突变体u-PA酶通过用纤溶酶-琼脂糖(Calbiochem)处理转化为相应于的双链酶。这些培养基中活性u-PA的浓度通过用标准PAI-1抑制剂制备物进行活性位点滴定而测量，所述标准PAI-1抑制剂制备物已预先如以上实施例1所述针对胰蛋白酶原始标准滴定。总酶浓度根据实验室手册Harlow et al(1998)Using Antibodies，Cold SpringHarbor Laboratory的方案用酶联免疫吸附测定而测量。这些浓度的比率产生在每种培养基中均有活性的u-PA变体的级分。
B.u-PA的直接生色测定
u-PA活性的直接测定利用底物苄氧羰基-L-γ-谷氨酰(α-t-丁氧基)-甘氨酰-精氨酸-对硝基酰苯胺单乙酸盐(Cbo-L-(γ)-Glu(α-t-βuO)-Gly-Arg-pNAAcOH；Spectrozyme
uPA，American diagnostica)(Madison et al.(1995)JBiol.Chem.，270：7558-7562)。通过测量在405nm吸光度每单位时间产生的游离(pNA)的吸光度的增加而确定酶活性。用6-8nM之间的酶浓度随时间进行动力学测定。SpectrazymeuPA的浓度在双链u-PA测定中在25到150μM之间变化，在u-PA的蛋白酶结构域的测定中在25到200μM之间变化。反应在96孔微滴板中进行，通过用Spectromax M2或M5读板器(Molecular Devices)每30秒测量405nm吸光度共测量2小时而评估反应速率。动力学常数kcat，Km和kcat/Km(特异性常数)通过将底物浓度的倒数对在OD405吸光度的速度的倒数作图并拟合进Lineweaver-Burk方程(1/速度＝(Km/Vmax)(1/[S])+1/Vmax；其中Vmax＝[E]*kcat)而计算。
下表13给出了在u-PA酶活性的直接测定中鉴别为对PAI-1/69抑制剂显示增加的敏感性的u-PA突变体的动力学分析结果。结果显示每种鉴别的突变体u-PA与野生型u-PA相比具有降低的酶活性，这通过测量对Spectrozyme
uPA底物的裂解的特异性常数(kcat/Km)而确定。在表中，测试的变体是上表11中在从u-PA噬菌体展示文库的后续代次(I至IV)选择后鉴别的那些。
C.用荧光底物进行的u-PA变体的动力学分析
用Ac-RRAR-AMC底物进行测量u-PA变体对RRAR靶底物序列的活性的直接测定。7-氨基-4-甲基香豆素(AMC)荧光肽底物的使用是确定蛋白酶特异性的常规方法(Zimmerman et al.(1977)Anal Biochem，78：47-51；Harris et al.(2000)PNAS，97：7754-7759)。酰苯胺键的特异性裂解释放了荧光AMC离去基团，提供了确定个体底物的裂解速率的有效手段。将底物系列稀释至0.05-12.0mM并在蛋白酶(9-25nM)存在下在Costar 96-孔半黑区(black half-area)测定平板中保温。来自游离AMC离去基团的荧光在荧光分光光度计(Molecular Devices Gemini XPS)中参照AMC标准在激发波长(380nm)和发射波长(450nm)测量。荧光增加速率在30分钟内以30秒间隔读取读数而测量。动力学常数kcat，Km和kcat/Km(特异性常数)通过将底物浓度的倒数对底物裂解速度的倒数作图并拟合进Lineweaver-Burk方程(1/速度＝(Km/Vmax)(1/[S])+1/Vmax；其中Vmax[E]*kcat)而计算。
下表14给出了u-PA变体ARF2和ARF36对Ac-RRAR-AMC底物的动力学分析的结果。结果显示所选择的u-PA蛋白酶变体对RRAR底物的特异性常数与野生型u-PA相比增加了大约或大于10倍。
D.用间接生色测定进行的纤溶酶原激活的动力学分析
进行间接生色测定以确定作为纯化的蛋白质制备物产生的野生型和突变体u-PA的活性(Madison et al.(1989)Nature，339：721-724；Madison et al.(1990)J Biol.Chem.，265：21423-21426)。在这一测定中，游离的对硝基苯胺通过由u-PA作用于纤溶酶原而产生的纤溶酶的作用而从生色底物Spectrozyme PL(H-D-正亮氨酰六氢酪氨酰-赖氨酸-对硝基酰苯胺二乙酸盐，American Diagnostics，Inc.)释放。游离的对硝基苯胺的释放在OD405nm经分光光度计测量。
为进行这一测定，将含有0.25-1ng待测的uPA酶，0.62mM SpectrozymePL，0.2μM Lys-纤溶酶原(American Diagnostics，Inc.)的100μl反应混合物与含有50mM Tris-HCL(pH 7.5)，0.1M NaCl，1.0mM EDTA和0.01％(v/v)Tween 80的缓冲液合并。反应在96孔平底微滴板(Costar，Inc.)中于37℃保温，在Molecular Devices Thermomax中每30秒测量405nm的光密度(OD405)，共1小时。如早先所述计算动力学常数kcat，Km和kcat/Km(特异性常数)(Madison，E.L(1989)Nature 339，721-724)。
下表15提供了在u-PA酶活性的间接测定中对鉴别为针对PAI-1/69抑制剂显示增加的敏感性的u-PA突变体的动力学分析的结果。结果显示每种鉴别的突变体u-PA与野生型u-PA相比具有降低的酶活性，这通过间接测量Spectrozyme
PL底物裂解的裂解特异性常数(kcat/Km)而确定。在表中，测试的变体是上表11中从相继代次(I至IV)的u-PA噬菌体展示文库中选择后而鉴别的那些。
E.野生型PAI-1和突变体PAI-1对突变体u-PA酶的抑制的动力学分析
用准一级(pseudo-first order)(ki＜2×106)或二级(ki＞2×106)条件确定突变体和野生型u-PA(阳性对照)的抑制的二级速率常数(ki)。对于每种酶，选择酶和抑制剂(突变体PAI-1)的浓度以产生若干数据点，其中残余酶活性在初始活性的20％至80％之间变化。在24℃在含有0.1mM EDTA和0.1％(v/v)Tween 20的0.1M Tris-HCl缓冲液(pH7.4)中进行野生型和突变体u-PA与野生型和突变体PAI-1的相互作用的速率的测量。使用上述D部分所述的间接生色测定以确定作为时间函数的残余酶活性。
PAI-1对野生型或突变体u-PA的抑制的速率常数是在准一级条件下，如前所述PAI-1相对于u-PA过量(参见例如Holmes et al.(1987)Biochemistry，26：5133-5140；Beatty et al.(1980)J.Biol.Chem.，255：3931-3934；Madison et al.(1990)PNAS，87：3530-3533；Madison et al.(1993)Methods Enzymol.，223：249-271)。简而言之，将纯化的野生型或突变体u-PA(3-50fmol)与野生型或突变体PAI-1(35-1330fmol)在室温保温0-120分钟。保温后，将混合物稀释并在如上述D中所述的标准生色测定中确定残余酶活性。通过对时间作图ln(残余活性/初始活性)并确定所得直线的斜率而分析数据。然后通过将斜率除以反应中抑制剂的浓度而获得准一级速率常数。
对于二级反应，将等摩尔浓度的野生型或突变体u-PA与野生型或突变体PAI-1在微滴板孔中直接混合并在室温预保温0-30分钟。预保温后，混合物用过量的中和抗PAI抗体猝灭并在间接生色测定中测量残余酶活性。将间接生色测定与不含PAI-1的或者在预保温和向反应混合物中加入抗PAI-1抗体、纤溶酶原和Spectrozyme PL后加入PAI-1的对照反应进行比较。通过对时间作图ln(残余活性/初始活性)并确定所得直线的斜率而分析数据。然后通过将斜率除以反应中抑制剂的浓度而获得二级速率常数。
下表16提供了PAI-1/RRAR抑制剂对u-PA变体的抑制的二级速率常数。结果表明变体ARF2和ARF36在对PAI-1/RRAR抑制剂底物的特异性方面改善20倍，这通过与野生型u-PA相比对于抑制的ki速率常数增加而评估。
下表17和18是野生型(表17)或突变体PAI-1/69抑制剂(表18)被针对PAI-1/69抑制剂选择的野生型u-PA或u-PA变体的抑制的二级速率常数的结果。表17和18中所示的变体u-PA是如以上表11所示的从一(I)至四(IV)轮相继的u-PA噬菌体文库选择鉴别的那些。表17的结果表明一些突变体u-PA(即u-PA/IIIa，u-PA/IIIb和u-PA/IVa)与野生型u-PA相比具有对于抑制稍稍增加的二级速率常数，突变体u-PA/Ic，u-PA/IIb和u-PA/IVb与野生型u-PA相比具有对于抑制降低的二级速率常数。表18的结果表明突变体PAI-1/69抑制剂对每种选择的u-PA变体的抑制的二级速率常数显著增加。结果表明每种选择的变体在对PAI-1/69抑制剂底物的特异性方面改善13倍以上，变体u-PA/IIIb，u-PA/IVa和u-PA/IVb各自显示出在特异性方面接近或超过40倍的改善。
实施例6
针对突变体AT3抑制剂从MT-SP1噬菌体文库选择变体MT-SP1
在野生型AT3的反应活性环(RSL)残基P4-P3-P2-P1-P1’-P2’中含有相应于氨基酸残基IAGRSL(SEQ ID NO：478)的六肽序列的突变体抗凝血酶III(AT3)抑制剂(SEQ ID NO：5)被突变以在这些残基中含有取代变为SLGRKI(SEQ ID NO：479)，相应于补体C2裂解序列的氨基酸残基。突变体AT3SLGR-KI在蛋白酶选择实验中用作“诱饵底物”以从大的组合MT-SP1噬菌体展示文库中分离对突变体底物序列有改善的反应性的噬菌体。简而言之，为分析第一代选择，将5μl 1∶100SM1(～3X1013)MT-SP1噬菌体文库(其为具有酶活性的低诱变频率文库(即0.2-0.5％诱变频率))以等表现度与5μl 1∶100SM2(～3×1012)MT-SP1噬菌体文库(其含有较高诱变频率(即0.9％))组合。将这些噬菌体文库与5μl肝素(5ng/μl；来自猪小肠粘膜原液)，5μl诱饵AT3SLGR-KI底物(浓度范围为0(即5μl H2O)，0.018μM，0.18μM，1.8μM或18μM)在5μl(18μM)野生型血浆纯化的AT3和5μl 10X MTSP活性缓冲液(0.5M Tris HCl，pH 8，0.3M NaCl，0.1％Tween 30)存在下以总体积50μl(即反应含有20μl H2O)混合。反应在37℃保温4.5小时。
MTSP-1噬菌体-AT3抑制剂复合物用CuSO4活化的琼脂糖捕获。200μl螯合琼脂糖(Pharmacia)用CuSO4(100mM)预活化。CuSO4活化的琼脂糖(100μl)用2ml于PBS缓冲液中的0.5％BSA在室温封闭1小时。通过在6500rpm沉淀60秒从封闭溶液中收获珠，随后重悬于450μl结合缓冲液(0.5M NaCl，100M Tris pH 8，10mM咪唑，0.1％Tween 20)中。将50μl上述淘选混合物反应加到CuSO4活化的琼脂糖珠以捕获His标记的AT3抑制剂-MTSP-1噬菌体复合物。在室温继续保温1小时。接下来在2000rpm离心淘选混合物1分钟，用500μl结合缓冲液洗涤含有结合的AT3噬菌体-MTSP-1抑制剂复合物的琼脂糖珠以除去未结合的噬菌体。重复洗涤步骤5次，在每次洗涤后将含有结合的复合物的珠转移到新管中以避免任何非特异性噬菌体的潜在“携带污染”。
结合的MT-SP1噬菌体-AT3SLGR-KI复合物在100μl洗脱缓冲液(0.5MEDTA，pH 8.0)中洗脱。50μl等份的洗脱的噬菌体用于在37℃感染3ml TG1大肠杆菌活跃生长细胞(A600＝0.5；0.5OD＝～1.5X108菌落/ml)20分钟。感染的细菌在含有羧苄青霉素(75μg/ml)的大平板(245mmX245mm)上铺板并在30℃保温过夜。第二天早晨收获平板。计数每个平板上的菌落并与不含AT3SLGR-KI抑制剂的背景平板进行比较。菌落计数结果如表19所述。
在平板被置于冷房间中2小时以固化琼脂后，将来自含有AT3SLGR-KI的平板的菌落刮下置于补加有羧苄青霉素(～100μg/ml)的25ml 2YT中。将20ml含有细菌的2YT培养基加入到500ml含有羧苄青霉素的2YT中，测定A600为0.13。将细菌生长至OD＝～0.5，然后加入～1X1010至2.6X1013cfu/ml的辅助噬菌体(VS M13)(在～150μl-200μl中)以扩增文库。在37℃振荡生长1小时后，培养物补加卡那霉素至终浓度为3μg/mL并在30℃振荡生长过夜。收获细胞并用PEG-NaCl溶液沉淀上清中存在的噬菌体颗粒。为分析第二代MT-SP1噬菌体选择，条件与第一代类似，只是针对AT3SLGR-KI诱饵底物的反应仅进行27分钟而不是4.5小时以增强选择的严格性。
在如上所述洗脱结合的MT-SP1噬菌体-AT3SLGR-KI复合物之后，用50μl等份的洗脱的噬菌体在37℃感染3ml TG1大肠杆菌活跃生长细胞(A600＝0.5；0.5OD＝～1.5X108菌落/ml)20分钟。感染的细菌在含有羧苄青霉素(75μg/ml)的大平板(245mm×245mm)上铺板并在30℃保温过夜。第二天早晨收获平板。计数每个平板上的菌落并与不含AT3SLGR-KI抑制剂的背景平板进行比较。菌落计数结果如表20所述。
挑取菌落以进一步鉴定。细胞在2ml补加羧苄青霉素(100μg/ml)和四环素(10μg/ml)的2xYT中生长，如所述进行噬菌体制备(Sambrook，J et al(1989)Molecular Cloning，A laboratory manual，Cold spring Harbor laboratory)。测试选择的噬菌体对Ac-SLGR-ACC底物的酶活性。另外，对选择的噬菌体根据对由野生型AT3或血浆所致失活的抗性进行选择。
实施例7
突变体AT3抑制剂的表达和纯化
A.变体AT3的产生
通过使用人抗凝血酶III(AT3)基因(SEQ ID NO：612，购自OrigeneTechnologies，Catalog # TC110831)的编码区作为模板将修饰导入AT3反应中心环(RCL)的氨基酸序列中而产生用作蛋白酶捕获“诱饵底物”的突变体AT3蛋白质。使用正向引物和反向引物经PCR扩增AT3 cDNA，所述正向引物具有SEQ ID NO：626所示的核酸序列：
CACGGGAGCCCTGTGGACATC(其含有填充序列(以上粗体显示)、BamH1位点(斜体显示)和与AT3基因杂交的部分(正常字体形式))，所述反向引物具有SEQ ID NO：628所示的核酸序列：GTAGCCAACCCTTGTGTTAAGGGAGGCGGAAGCCATCACCACCATCACCACTAAGAATTC。在扩增后，使用限制位点Bam和EcoRI将cDNA亚克隆进pAcGP67b杆状病毒转移载体(BD Biosciences SEQ ID NO：494)。在这一亚克隆步骤中加入C末端6xHis标记(SEQ ID NO：496)或C末端FLAG标记(DYKDDDDK；SEQ ID NO：495)以便AT3突变体后来可以通过亲和层析分离。克隆的AT3融合蛋白的核苷酸和氨基酸序列分别如SEQ ID NO：613和614所示，其含有用4个氨基酸GGGS接头(SEQ ID NO.：620)融合于6xHis标记的AT3。
为制备突变体AT3诱饵底物以分离具有各种特异性的靶蛋白酶，使用Quikchange
定点诱变试剂盒(Stratagene)根据供应商规定的条件进行诱变反应以导入靶裂解序列的氨基酸残基来替代野生型AT3反应中心环(RCL)序列IAGRSL(SEQ ID NO.：478)(SEQ ID NO.：5所示前体AT3多肽序列的氨基酸残基422-427；以及SEQ ID NO.：493所示成熟AT3多肽序列的氨基酸残基390-395)。
一种这样的突变体AT3(AT3/RRVR-KE)(SEQ ID NO.：497)是通过用来自靶向的VEGFR2裂解序列的氨基酸残基RRVRKE(SEQ ID NO：498)置换野生型IAGRSL氨基酸序列的氨基酸残基而制备的。另一种突变体AT3(AT3/SLGR-KI)(SEQ ID NO：499)是通过用来自靶向的补体C2蛋白裂解序列的氨基酸残基SLGRKI(SEQ ID NO：479)置换野生型IAGRSL氨基酸序列而制备的。
对于Quikchange
PCR，野生型AT3RCL引物具有如下核酸序列，如SEQ ID NO：630所示：GCTGCAAGTACCGCTGTTGTGATTGCTGGCCGTTCGCTAAACCCCAACAGGGTGACTTTC。补体C2靶序列引物具有如下核酸序列，如SEQ ID NO.：632所示：GCTGCAAGTACCGCTGTTGTGTCGTTAGGCCGTAAAATTAACCCCAACAGGGTGACTTTC。VEGFR2靶序列引物具有如下核酸序列，如SEQ IDNO.：634所示：GCTGCAAGTACCGCTGTTGTGCGCCGTGTGCGCAAAGAAAACCCCAACAGGGTGACTTTC。
含有野生型AT3cDNA的载体和含有突变体AT3cDNA的载体各自转化XL-1-Blue超感受态细胞(Stratagene)并在其中扩增。用Qiagen PlasmidMaxi试剂盒(Qiagen)根据供应商规定的条件从细胞纯化质粒DNA。
B.AT3突变体的表达
使用上述含AT3的pAcGP67b转移载体用Sf9昆虫细胞表达和纯化His标记的和FLAG标记的野生型和突变体AT3蛋白。Sf9细胞在SF900 II无血清培养基(Invitrogen)中适应生长并在35mm培养皿中生长至85-90％铺满。细胞用FlashBac
杆状病毒表达系统(Oxford Expression Technologies)根据供应商规定的条件和方案转染。将500ng AT3转移载体和500ngFlashBac
重组载体在1ml不含抗生素的SF900II无血清培养基中与5μlCellfectin
转染试剂(Invitrogen)预保温20分钟，然后滴加到细胞。转染5小时后，离心细胞并重悬于2mL含有抗生素(抗生素/抗真菌剂溶液；Cellgro)的SF900II无血清培养基中，在28℃保温4天。病毒在Sf9细胞中扩增至最大效价为1×106pfu/mL，这通过噬斑测定确定。然后用HighFive(BTI-TN5B 1-4)昆虫细胞系(Invitrogen)和ExcellTM405无血清培养基(JRH Biosciences)表达重组AT3。细胞以感染复数(MOI)为0.1-1感染并在1L Erlenmeyer培养瓶中的300mL培养体积中生长4-5天，所述培养瓶在轨道摇床上以125RPM振荡。
C.基于亲和性纯化野生型和突变体AT3蛋白
为进行His标记的AT3蛋白的基于亲和性的纯化，将来自实施例7B的培养物上清经离心和用0.45μM滤膜过滤而澄清，并透析进含有50mM磷酸钠pH 7.5，300mM NaCl的缓冲液中。用BioLogic DuoflowTM层析装置(Bio-Rad)和10mL TALON
钴金属亲和树脂(Clontech)经柱层析纯化蛋白质。用50mM磷酸钠pH 7.5，300mM NaCl和50mM磷酸钠pH 6.5，300mMNaCl，150mM咪唑的线性梯度洗脱树脂结合的His标记的蛋白质。合并含有蛋白质的级分并透析进AT3储存缓冲液(50mM磷酸钠pH 6.5，300mMNaCl，5％甘油)中。为证实纯化的AT3制备物含有活性蛋白质，进行如以下实施例14所述的MT-SP1抑制(活性位点滴定)测定以测量透析的AT3抑制MT-SP1裂解底物的能力的能力。在SpectraMax
M5(SpectraMax
M5Microplate Reader，Molecular Devices)(Molecular Devices，Inc)上动力学评价来自这一MT-SP1抑制测定的反应混合物对0.4mM Ac-RQAR-ACC(乙酰-Arg-Gln-Ala-Arg-ACC)底物(商业合成)的裂解。这一底物名称中的“ACC”是指7-氨基-4-氨基甲酰甲基香豆素离去基团。ACC离去基团在激发波长(Ex)＝380，发射波长(Em)＝450和截断值波长(c/o)＝435检测。纯化的His标记的AT3蛋白的总收率约为1-3mg/L。
为进行FLAG标记的AT3蛋白的基于亲和性的纯化，如上所述经离心和过滤澄清来自培养物的上清。澄清的上清透析进Tris缓冲盐水(TBS)pH 7.4中并以300mL总体积加到新鲜的1L Erlenmeyer培养瓶中。加入2mL预平衡的抗FLAG M2亲和凝胶(Sigma)并将总体积在轨道摇床上以125RPM在4℃保温3小时。用多孔20mL层析柱(Bio-Rad)经重力收集树脂结合的FLAG标记的AT3蛋白。树脂先用5mL TBS洗一次，用5mL TBST(含有0.1％Tween-20的TBS)洗一次，用5mL TBS洗第二次。然后AT3蛋白通过加入10mL含有0.2mg/mL FLAG肽(Sigma)的TBS洗脱。浓缩洗脱液并透析进AT3储存缓冲液(50mM磷酸钠pH 6.5，300mM NaCl，5％甘油)，使用上述以及以下实施例14A中所述的matriptase(MT-SP1)抑制(活性位点滴定)测定如上所述测定活性。纯化的FLAG标记的AT3蛋白的总收率大约为0.5-1mg/L。
这些纯化的FLAG和His标记的AT3蛋白例如在以下实施例描述的方法中用作蛋白酶捕获剂以鉴别识别特定靶位点序列的蛋白酶。
实施例8
蛋白酶变体的噬菌体展示文库的构建
A.将野生型和C122S MT-SP1蛋白酶结构域(B链)克隆进pMal-C2噬菌粒展示载体
使用限制位点Nde1和HindIII将编码含有SEQ ID NO：253所示全长MT-SP1蛋白的氨基酸615-854的成熟MT-SP1蛋白酶结构域(MT-SP1 B链)(SEQ ID NO：505)的cDNA(SEQ ID NO：504)克隆进pMal-C2载体(SEQID NO：615)(New England Biolabs)，该载体含有促进分泌的STII前导序列(TGAAAAAGAATATCGCATTTCTTCTTGCATCTATGTTCGTTTTTTCTATTGCTACAAACGCGTATGCA(SEQ ID NO.：636)和编码丝状噬菌体M13GeneIII(SEQ ID NO：616)的C末端结构域的核酸。MT-SP1蛋白酶结构域cDNA插入前导序列和GeneIII结构域之间以便最终构建体含有SEQ ID NO：510所示的核酸序列，其编码MT-SP1N末端GeneIII融合蛋白。这一融合蛋白的编码氨基酸序列(SEQ ID NO：506)如下所示，STII前导序列(SEQID NO：511)正常字体显示，成熟MT-SP1结构域粗体显示，C末端GeneIII(SEQ ID NO：512)结构域斜体显示。*表示在编码蛋白质的核酸中存在的终止密码子。
SEQ ID NO.：506：
mkkniafllasmfvfsiatnaya
除了野生型MT-SP1B链融合蛋白之外，使用上述方法还产生了MT-SP1 B链变体(CB469)-GeneIII融合蛋白。SEQ ID NO：507所示的CB469变体氨基酸序列通过用丝氨酸取代野生型MT-SP1蛋白酶结构域序列的第122位(基于胰凝乳蛋白酶编号)的半胱氨酸而产生，所述野生型MT-SP1蛋白酶结构域序列如上述斜体序列所示并示于SEQ ID NO：505。这一CB469序列被克隆进上述pMal-C2载体中。为了获得改良的展示，使用含有编码这一变体MT-SP1融合蛋白的核酸的噬菌粒载体如以下实施例8B所述产生MT-SP1突变体噬菌体展示文库。
B.诱变蛋白酶结构域以产生突变体噬菌体展示文库
使用如下举例示出的本领域已知的标准易错PCR诱变方案产生含有突变的蛋白酶结构域的噬菌体展示文库(Matsumura et al.，Methods Mol Biol.2002；182：259-67；Cirino et al.，Methods Mol Biol.2003；231：3-9)。
1.B链MT-SP1融合蛋白的诱变
为了构建含有突变MT-SP1的融合蛋白，通过易错PCR从pMal-C2载体扩增MT-SP1 CB469cDNA，所述易错PCR使用DiversifyPCR随机诱变试剂盒(BD Biosciences，Clonetech)并参照供应商建议的条件，每千碱基获得5个突变。用于这一PCR的具有SEQ ID NO：508)所示核酸序列GCGCAGATATCGTACCG
的MT-SP1正向引物设计用于在STII前导序列内杂交(残基以斜体示出)并含有Nde限制位点序列(粗体示出)。具有SEQ ID NO：509所示核酸序列GTGCATGCTGACTGACT的MT-SP1反向引物设计用于在编码MT-SP1蛋白酶结构域的序列的3’部分杂交(残基以斜体示出)并含有Sac1限制位点序列(粗体示出)。
2.诱变产物的纯化
Taq聚合酶与DNA紧密结合并因此未被Qiagen PCR纯化试剂盒完全除去；其存在可能干扰下游PCR产物的限制消化(Crowe et al.，Nucleic AcidsRes.1991 January 11；19(1)：184；Wybranietz et al.，Biotechniques(1998)24，578-580)。因此，为了从来自实施例8B(1)的扩增的野生型和突变体PCR产物中除去Taq聚合酶，在它们纯化之前，向反应中加入下列物质：5mMEDTA、0.5％SDS、50ng/μl蛋白酶K。为了消除Taq聚合酶，将混合物在65℃保温15分钟。
为保证没有剩余野生型模板(其可能潜在干扰下述选择方法)，纯化PCR产物以除去模板DNA。为从PCR产物中分离载体，在不与PCR产物或梯共迁移的10X Orange G凝胶上样缓冲液(New England Biolabs)存在下将样品上样至1％琼脂糖凝胶上。用解剖刀将PCR产物从凝胶上切下来。用QIAquick
凝胶提取试剂盒方案(Qiagen)或ZymocleanTM凝胶提取试剂盒(Zymoclean CA，Cat#D4001)根据供应商指明的条件纯化切下来的产物。
对于Qiagen凝胶提取试剂盒，将切下的凝胶片段在QG缓冲液中溶解并缓慢流过Qiagen QG柱，每个均具有大约5-10μg的结合能力，并且保留不超过2mL。使用足够数量的柱以容纳起始材料的全部体积。在收集试管中以14000rpm离心柱以除去任何残留的缓冲液QG。将0.7mL缓冲液PE加入到每个柱中并保温样品2-5分钟。然后在13000rpm离心柱2次，每次1分钟，以除去所用残余PE缓冲液。将样品转移至新的1.5mL微离心管中，随后加入50μL H2O并保温2分钟。结合的DNA经在7000rpm离心洗脱。典型收率为样品起始量的30％-60％之间。
对于ZymocleanTM凝胶提取试剂盒，根据起始材料的量，使用一或多个ZymocleanTM凝胶提取试剂盒柱(每个柱具有5μg最大结合能力)或者一或多个来自ZymocleanTM DNA Clean & ConcentratorTM试剂盒的柱(每个柱具有25μg能力)。使用这一试剂盒，切下的DNA片段被转移到1.5ml微离心管中，随后向从凝胶切下的每个体积的琼脂糖中加入3个体积的ADBBufferTM。样品在37-55℃保温5-10分钟直至凝胶片完全溶解。溶解的琼脂糖溶液被转移到收集试管中的Zymo-Spin ITM柱中并在至少10000rpm离心30-60秒。弃掉来自柱的流经液。将200μL洗涤缓冲液加入柱中并在至少10000rpm离心30秒。弃掉流经液并重复洗涤步骤。将50μL水直接加到柱基质中。对于ZymocleanTM凝胶提取柱最小洗脱体积为10μL，对于DNAClean & ConcentratorTM柱是35mL。将柱放进1.5ml试管中并在至少10000rpm离心30-60秒以洗脱DNA。
用这两种方法之一洗脱后，合并样品并通过在70μL UV杯中用分光荧光计测量样品在260nm的吸光度而评估DNA浓度。DNA浓度根据下列等式计算：1A260＝50ng/μl ds DNA。
C.使用pMal-C2噬菌粒载体构建蛋白酶突变体噬菌体文库
为构建表达突变体蛋白酶结构域的噬菌体展示文库，将消化的PCR产物如得自实施例8A和B描述的蛋白酶PCR诱变和纯化的PCR产物连接进上述pMal-C2噬菌粒载体。为此，用Nde1和Sac1消化载体，将产物凝胶纯化并与纯化的限制消化的PCR产物在连接反应(下述)中合并。Nde1/Sac1消化的凝胶纯化的pSTII-g3 pMal-C2噬菌粒的分子量(MW)是5835碱基对(bp)；Nde1/Sac1消化的MT-SP1 PCR产物的MW是806bp。典型地，对于2mL连接反应，7.58μg切割载体是大约1nM载体，3.14μg切割MT-SP1产物是大约3nM插入产物。
对于MT-SP1产物的连接，40μl(3nM，3.14μg)消化的纯化的产物与以下物质混合：40μl(1nM，7.58μg)消化的纯化的载体；1510μl H20；400μl 5X T4DNA连接酶缓冲液(Gibco)；和10μl(10单位)T4DNA连接酶(New EnglandBiolabs)。连接反应在2mL体积中在16℃进行过夜或者在室温进行4小时。连接后，将连接反应混合物在65℃保温15分钟而热失活DNA连接酶，随后加入4mL ZymoResearch DNA结合缓冲液。然后每次800μl将这一样品加入到25μg ZymoResearch柱中。柱用600μl ZymoResearch洗涤缓冲液洗2次并用预热至42℃的50μL水洗脱。通过用分光荧光计测量稀释的洗脱样品(3μl洗脱在70μl H2O中)在260nm的吸光度而评估DNA回收百分比。例如，对于上述3mL连接，如果A260相应于0.12＝140ng/μl，则总收率经确定为大约7μg(其为从连接中大约70％的DNA回收)。
将连接产物电穿孔进XL-1blue细胞，通过电穿孔其可容纳大约500ng/μl。每200μl XL-1Blue细胞(Stratagene)加入7.5μl 140ng/μl连接液。细胞加入到0.2cm间距杯中并在Gene Pulsar(Bio-Rad，CA)中使用下述条件电穿孔：电压(V)＝2500，电容＝25(uF)，电阻＝200欧姆。在电穿孔后立即将1mL SOC培养基(Invitrogen)加到杯中。然后将细胞转移至25mL SOC培养基中并在37℃保温20分钟。保温后，制备细胞的100倍系列稀释液的5个少量等份(100微升)并在小2YT羧苄青霉素琼脂平板上铺板，在37℃保温过夜以计数菌落数(代表来自由电穿孔产生的文库的克隆数)。在一种方法中，剩余的培养物体积被离心以沉淀细胞，随后重悬于12mL SOC缓冲液中并在补加羧苄青霉素(75ug/ml)的大琼脂平板(245mmX245mm)上铺板，在30℃生长过夜。或者，为从文库制备噬菌体原液，将细胞直接加入到具有1×1010CFU/mL的M13KO7辅助噬菌体的补加了羧苄青霉素(75ug/ml)的500mL 2YT培养基中并在37℃生长过夜。
实施例9
使用突变体AT3蛋白酶捕获剂从蛋白酶结构域噬菌体文库中选择变体蛋白酶结构域
A.选择MT-SP1噬菌体/突变体AT3复合物及基于ELISA的读出测定
在本实施例中，如上述实施例7所述含有补体C2靶裂解序列SLGRKI的突变体AT3抑制剂在淘选实验中被用作“诱饵底物”，该实验被设计用于分离和提供对靶裂解序列具有改良的反应性的携带突变体MT-SP1的噬菌体的读出。用下述程序从实施例8A、8B和8C所述的大的组合MT-SP1噬菌体展示文库中分离噬菌体。
1.携带突变体MT-SP1的噬菌体与AT3变体相互作用并裂解AT3变体
来自噬菌体文库的噬菌体结合的MT-SP1突变体首先用具有如下靶裂解位点的突变体AT3选择。通过在70μL体积中保温下述反应成分而在96孔聚苯乙烯平板(Nunc Maxysorp)的一式二份孔中在37℃进行每个裂解反应60分钟：35μl MT-SP1文库噬菌体，3.14E12CFU/ml；7μL 10X MT-SP1活性缓冲液(0.5M Tris-HCl，pH 7.4，1M，NaCl，1％Tween20)；7μL 10μM低分子量肝素(BD Biosciences)；14μL H2O；和7μL His标记的突变体AT3-SLGRKI。用以下不同的AT3-SLGRKI浓度进行各个裂解反应：100nM，33nM，11nM，3.3nM，1.1nM和0.33M。通过加入2μL 100mg/ml蛋白酶抑制剂4-(2-氨基乙基)-苯磺酰氟(Pefabloc，Roche Diagnostics)而终止每个反应。然后加入40μL 0.55％BSA，0.275％Tween 20溶液并通过上下吸取而充分混合。
2.用抗His抗体捕获MT-SP1噬菌体-AT3复合物
同时，另一96孔聚苯乙烯平板(Nunc Maxysorp)的孔用100μL于0.2M碳酸盐缓冲液(pH 9，Pierce)中的5ng/μl链霉抗生物素(Pierce)在室温振荡包被1小时。孔然后用250μL PBST洗3次，用200μL于PBS中的0.2％BSA在室温封闭2小时，与100μl 5ng/μl生物素化抗6HIS抗体在室温保温1小时以捕获His标记的AT3突变体，并用PBST充分洗涤。然后来自实施例9A(1)中的AT3裂解反应的每种突变体MT-SP1噬菌体样品以一式二份加入到包被的96孔板中并在室温保温1小时。平板然后用250μL PBST洗14次。
3.MT-SP1噬菌体-AT3复合物的选择的基于ELISA的读出
洗涤后，微滴板2排中的第1排用于进行基于ELISA(酶联免疫测定)的测定以获得噬菌体捕获的读出。向这一排中的每孔中加入100μL 1∶5000稀释的HRP缀合物抗M13噬菌体抗体(GE Healthcare)并进行结合1小时。这一排中的孔然后用Skanwasher平板洗涤器(Molecular Devices)洗8次，随后加入100μL TMB/过氧化物底物溶液(Pierce)并在室温保温5分钟。这一反应用100μL 2M H2SO4猝灭并在SpectraMax平板读数器(MolecularDevices)上测定450nM的吸光度。这一读出用作存在噬菌体-AT3复合物的代表(surrogate)。在这一测定中，观察到吸光度的浓度依赖性增加(基于增加实施例9A(1)中的裂解反应中所用的AT3-SLGRKI诱饵的浓度)。另外，当用本文所述的连续轮次淘选进行这一过程时，在每轮后观察到增加的吸光度，表明这一淘选方法可以针对靶裂解位点亲和性连续富集噬菌体库。这些数据提示这一方法可以用于从噬菌体文库选择对特定靶序列具有亲和性的突变体蛋白酶结构域融合蛋白。使用用于基于亲和性的捕获和以下实施例9A(4)所述的后续洗脱的相同96孔平板，ELISA测定提供了用于获得这一选择和富集的读出的方法。
4.选择的MT-SP1噬菌体的洗脱
来自实施例9A(2)的微滴板的两排中的第二排然后被用于特异性洗脱结合的噬菌体以用于后续的纯化和筛选。向这一排的每孔中加入100μL100mM HCL并保温5分钟以洗脱特异性结合的噬菌体。将所得噬菌体洗脱液加入到含有33μL 1M Tris pH8的单独的孔中以用于在感染前进行酸中和。然后将133μL中和的噬菌体混合物加入到生长至OD600为0.5的1mlXL-1 Blue细胞，细胞然后在37℃保温20分钟并铺板在补加羧苄青霉素的2xYT琼脂平板(245mm×245mm)上。这些细胞用于后续的筛选、测序和纯化方法中，如以下实施例所述。
这一选择和基于ELISA的测定方法还用于选择和评估来自如以上实施例2和3所述的uPA文库的uPA突变体。
B.携带选择的蛋白酶结构域的噬菌体的纯化
本实施例描述了分离已用诱饵抑制剂选择的噬菌体上清的方法。确定了选择的噬菌体如上述实施例9A回收的那些的效价(cfu/mL)。用PBS稀释噬菌体原液并用于感染生长为OD600为0.5的或大约2.1×108细胞/mL的大肠杆菌XL-1 Blue细胞。希望的感染性范围为每平板1000-2000个菌落。感染的细胞铺板在方形245mm×245mm聚苯乙烯BioAssay(Corning)培养皿中的补加100μg/mL羧苄青霉素的2YT琼脂上并生长16-20小时或者直至菌落大小大约为直径2mm。使用自动化菌落挑取器挑取个体菌落并分散在96孔聚丙烯平板的孔中，每孔含有150-170uL补加100μg/mL羧苄青霉素和12μg/mL四环素的2YT培养基。对照孔用细胞接种，所述细胞用模板噬菌体(具有已被用作诱变模板的蛋白酶结构域的噬菌体)或用含有融合蛋白(其含有无活性蛋白酶变体)的噬菌体感染。接种的平板用透气膜密封，置于HiGro(GeneMachines)培养箱并在37℃以400rpm振荡14-20小时。
保温后，为获得log期细胞，从每孔取100μL用于接种含有1mL补加100ug/mL羧苄青霉素和12μg/mL四环素的2YT培养基的深孔96孔板的孔。深孔板然后用透气膜密封并置于HiGro
培养箱，在37℃以400rpm有氧振荡直至细胞密度达到OD600为0.4-0.6之间。典型的培养时间为4-5小时之间。保温后，向每孔中加入100μL辅助噬菌体原液，密封平板并再在37℃以400rpm振荡5-10分钟。振荡5-10分钟后，平板不振荡在37℃静态保温30-45分钟。然后重新以400rpm在37℃振荡15-30分钟。振荡后，向每孔加入100μL卡那霉素溶液(400μg/mL)以在每孔中产生33.3μg/mL的终浓度。再次密封平板，并在37℃以400rpm振荡12-16小时。为沉淀细胞，平板然后在4℃以3500-4500rpm离心20分钟。离心后，含有分离的噬菌体的上清立即用于如以下实施例所述的筛选，或者先储存在4℃。
C.蛋白酶结构域噬菌体上清的聚乙二醇(PEG)沉淀
本实施例描述了用聚乙二醇沉淀在纯化的选择的噬菌体上清中除去潜在污染性背景蛋白酶活性(本文下述的一些鉴定测定对于所述蛋白酶活性是敏感的)的方法。在这一方法中，在用辅助噬菌体过夜(12-16小时)拯救携带MT-SP1的噬菌体上清(如在实施例9A中选择和洗脱的那些)后，样品在4℃以3500-4500rpm离心20分钟，从每孔中取1000μL上清并转移到另一96孔深孔板的孔中。
为了沉淀，向每孔中加入250μL含有在2.5M NaCl中的20％PEG(体积)的溶液。密封平板并通过剧烈颠倒混合，然后置于冰水浴中静止1-2小时。平板然后在4500rpm离心60分钟或在3500rpm离心90分钟。倾倒出每孔的上清溶液，将平板拍干并排水20-30分钟。所得沉淀用PBS重悬至终体积相当于原始噬菌体上清体积(200uL)的20％，以获得5倍浓缩液。这一材料立即用于下列测定或者储存在4℃直至测试。
实施例10
对靶底物序列具有增加的反应性和催化效率的携带蛋白酶结构域的噬菌体的筛选
如实施例9B和9C所述的个体噬菌体制备物用于各种测定中以确定它们的特异性和/或活性。
A.通过监测诱饵蛋白对荧光肽水解的抑制而分析表达蛋白酶结构域变体的噬菌体
作为一种评估携带突变体蛋白酶的噬菌体克隆的方法，可以进行生物化学抑制测定以比较抑制剂(诱饵丝氨酸蛋白酶抑制蛋白)抑制选择的变体蛋白酶结构域的活性的能力及其抑制最初用于噬菌粒文库构建中的模板蛋白酶结构域(即“亲代”蛋白酶)的活性的能力。使用这一方法，在存在和不存在给定浓度的抑制剂诱饵的情况下评估携带突变体蛋白酶的噬菌体(如上述实施例9回收的那些)裂解含有靶底物序列的荧光底物的能力，并与模板蛋白酶结构域在存在相同诱饵下裂解相同序列的能力进行比较。使用荧光肽底物是用于确定蛋白酶特异性的常规方法(Zimmerman et al.(1977)Anal Biochem，78：47-51；Harris et al.(2000)PNAS，97：7754-7759)。
为了分析与MT-SP1 B链模板(用于实施例8B(1)中的突变)的抑制作用相比较的MT-SP1 B链突变体的抑制作用，可以使用变体AT3(具有希望的靶序列)作为抑制剂，Ac-RQAR-ACC可以用作底物。在这一底物中，酰苯胺键的裂解释放了荧光ACC离去基团，提供了确定针对各个底物的裂解速率的有效手段。
在这种测定的一个实例中，携带uPA蛋白酶的噬菌体如上述实施例3中回收的那些噬菌体裂解荧光底物Ac-AGR-AMC(SEQ ID NO：617)的能力在存在或不存在给定浓度的PAI诱饵的情况下被评估。如上所述，使用这种7-氨基-4-甲基香豆素(AMC)荧光肽底物是确定蛋白酶特异性的常规方法。在这一实例中，将35μL噬菌体上清(如在实施例3(A)中获得的)转移到384孔聚丙烯平板(CoStar，#3658)的指定的测定孔和指定的对照孔中。向测定空中加入35μL含有用于选择中的相同PAI诱饵的2x间接测定缓冲液。诱饵浓度与选择中使用的相同。向相应的对照孔中加入35μL 2x间接测定缓冲液(无抑制剂)。平板在37℃保温60分钟。将噬菌体与抑制剂或对照缓冲液混合后，向孔中加入10μL在间接测定缓冲液中稀释至终测定浓度为60μM的AGR-AMC荧光肽底物。用Molecular Devices SpectraMax
Plate Reader使用动力学读数模式在380nm激发波长和460nm发射波长测量荧光1小时。对于模板蛋白酶而言，显示增强的靶底物抑制的克隆被进一步分析。
B.用荧光肽底物分析变体蛋白酶活性
为直接评估MT-SP1突变体的活性和特异性，用荧光肽底物Ac-RQAR-ACC(具有由野生型MT-SP1识别的天然自催化裂解序列)和Ac-SLGR-ACC(乙酰基-Ser-Leu-Gly-Arg-ACC)(具有C2靶位点裂解序列)进行了测定。如上所述，这些底物名称中的ACC代表7-氨基-4-氨甲酰基甲基香豆素，其是荧光离去基团。还如上所述，使用荧光肽底物是确定蛋白酶特异性的常规方法(Zimmerman et al.(1977)Anal Biochem，78：47-51；Harris et al.(2000)PNAS，97：7754-7759)。在这个实例中，酰苯胺键的特异裂解释放了荧光ACC离去基团，提供了确定针对各个底物的裂解速率的有效手段。在这一方法中，35μL 2x间接测定缓冲液被加到所有测试孔中。向每个指定孔中加入35μL噬菌体上清(如实施例9B所述分离的)或重悬的PEG沉淀的噬菌体(如实施例9C所述分离的)。在加入噬菌体后，平板在2000rpm离心1分钟以除去气泡。10μl每种肽底物(Ac-SLGR-ACC(最终测定浓度＝125μM))和Ac-RQAR-ACC(最终测定浓度＝60uM))用1X间接测定缓冲液稀释，然后逐个加到合适的孔中。用动力学读数模式的SpectraMax
M5Microplate Reader(Molecular Devices)随时间监测水解速率(ROH)，以相对荧光单位/秒RFU/s)测量，是底物裂解的表示。
实施例11
MT-SP1突变体的产生、选择、评估和鉴别
A.来自噬菌粒文库的B链MT-SP1突变体的荧光测定
本实施例描述了一种荧光测定，其用于分析用如实施例8所述使用文库产生的携带MT-SP1蛋白酶结构域的噬菌体的活性和特异性。如以上实施例8所述使用MT-SP1的天然B链作为模板制备了MT-SP1文库，所述天然B链在基于胰凝乳蛋白酶编号的第122位具有丝氨酸代替半胱氨酸(SEQ ID NO：507)。用pMal-C2载体和供应商推荐的易错PCR条件如上所述制备了文库以实现大约0.5％的诱变率。来自这一诱变反应的收率是4×109个重组体。
基于与含有靶底物序列(代替天然RCL；如实施例7所述)的变体AT3相互作用并裂解其的速率选择噬菌体是如实施例9A所述用聚丙烯96孔形式进行。为这一选择，1×1012个重组噬菌体与3.3nM携带SLGRKI RCL序列的变体AT3混合30分钟。洗涤后，如以上实施例9A所述洗脱噬菌体，并如实施例9B所述用于感染XL-1 blue细胞。
使用本文提供和描述的方法进行连续轮次的选择以根据靶底物序列的快速相互作用和裂解进行富集。例如，第一轮中选择的克隆用于进行本文所述的第二轮选择，如实施例9A(3)所述使用3.3，1.1和0.33nMAT3进行1小时。
第一轮选择后，如实施例9C所述用PEG沉淀从选择的克隆制备噬菌体上清。用上述实施例10(B)的方法使用Ac-SLGR-ACC(含有C2靶裂解位点序列)和Ac-RQAR-ACC(含有天然MT-SP1的天然裂解序列)作为荧光底物筛选噬菌体克隆。对于每一克隆，从Ac-SLGR-ACC测定确定的荧光速率(ROF)与从Ac-RQAR-ACC测定确定的ROF进行比较以作为比较每种突变的MT-SP1蛋白酶结构域对天然靶底物序列的活性及其对靶底物序列的活性的手段。突变体MT-SP1测定中的ROF也与模板(C122S)MT-SP1测定中的ROF进行了比较。用各个克隆获得的结果如下表21所示，表中列出了克隆编号并列出了表示为RFU/s(相对荧光单位每秒)的荧光速率。
表21：针对AT3-SLGRKI的裂解速率选择的携带突变体MT-SP1蛋白酶结构域的噬菌体的筛选
| 突变体MT-SP1克隆编号 | Ac-SLGR-ACC速率(RFU/秒) | Ac-RQAR-ACC速率(RFU/秒) |
| 模板 | 1.85 | 15.6 |
| CPC-0019595 | 7.1 | 33 |
| CPC-0023085 | 0.8 | 2 |
| CPC-0023230 | 1.3 | 4 |
| CPC-0023401 | 3.9 | 12 |
| CPC-0023949 | 0.7 | 2 |
| CPC-0024129 | 3.8 | 15 |
| CPC-0024153 | 2.5 | 6 |
| CPC-0024527 | 4.3 | 12 |
| CPC-0024715 | 3.2 | 12 |
| CPC-0025366 | 1.3 | 1 |
| CPC-0025387 | 6.8 | 14 |
| CPC-0025533 | 6.9 | 23 |
| CPC-0025582 | 1.7 | 3 |
| CPC-0025720 | 2.5 | 5 |
| CPC-0025866 | 1.2 | 4 |
| CPC-0025876 | 4.0 | 8 |
| CPC-0025890 | 10.6 | 33 |
| CPC-0025941 | 1.0 | 4 |
| CPC-0025974 | 9.3 | 41 |
| 突变体MT-SP1克隆编号 | Ac-SLGR-ACC速率(RFU/秒) | Ac-RQAR-ACC速率(RFU/秒) |
| CPC-0026100 | 14.8 | 25 |
| CPC-0026122 | 6.5 | 31 |
| CPC-0026125 | 17.4 | 84 |
| CPC-0026200 | 7.0 | 21 |
| CPC-0026219 | 8.0 | 23 |
| CPC-0026232 | 7.1 | 15 |
| CPC-0026597 | 11.0 | 34 |
| CPC-0026727 | 0.8 | 2 |
| CPC-0026761 | 7.8 | 25 |
| CPC-0027290 | 3.9 | 12 |
| CPC-0027306 | 11.0 | 50 |
| CPC-0027309 | 8.3 | 50 |
| CPC-0027326 | 9.1 | 54 |
| CPC-0027335 | 12.3 | 57 |
| CPC-0027369 | 2.3 | 11 |
| CPC-0027399 | 13.4 | 99 |
| CPC-0027484 | 2.5 | 12 |
| CPC-0027516 | 3.4 | 17 |
| CPC-0027617 | 1.4 | 7 |
| CPC-0027706 | 0.4 | 1 |
| CPC-0027718 | 2.1 | 7 |
| CPC-0027797 | 5.5 | 15 |
| CPC-0027841 | 2.7 | 9 |
| CPC-0028017 | 5.1 | 16 |
| CPC-0028333 | 5.6 | 17 |
| CPC-0028341 | 5.5 | 26 |
B.通过DNA测序鉴别选择的MT-SP1突变体噬菌体
本实施例描述了用于鉴别如前述实施例所述制备并基于如实施例10B所述的荧光测定的结果而选择的阳性噬菌体克隆的方法。为此方法，各个克隆与XL-1Blue大肠杆菌细胞混合以用于感染，培养物在37℃振荡生长过夜。用质粒制备试剂盒(Qiagen)从过夜培养物纯化质粒DNA，并送出DNA进行测序以鉴别突变体。
在这种方法的一个实例中，来自上述实施例11A的选择的B链MT-SP1突变体的氨基酸序列用上述步骤鉴别。用于鉴别这些克隆的测序引物示于SEQ ID NO：618：5’GGTGTTTTCACGAGCACTTC3’。分析测序数据获得的结果见下表22。表中仅列出了具有在一个以上的分离物中发现被突变的残基的突变体。表22列出了每一克隆与野生型MT-SP1 B链序列(SEQ ID NO：505)相比的氨基酸突变/位置，其用测序数据分析而确定。氨基酸编号根据胰凝乳蛋白酶编号。还列出了SEQ ID NO表示编码含有指明的氨基酸突变的MT-SP1蛋白酶结构域(B链)的氨基酸残基的序列以及编码具有相同突变的全长MT-SP1蛋白的氨基酸残基的序列。
表22：选择的MT-SP1突变体
| 突变体MT-SP1克隆编号 | 氨基酸突变(胰凝乳蛋白酶编号) | SEQ ID NO(蛋白酶结构域)： | SEQ ID NO(全长)： |
| CPC-0019595 | C122S/I136T/N164D/T166A/F184(A)L/D217V | 516 | 537 |
| CPC-0023085 | I41F/C122S | 517 | 538 |
| CPC-0024153 | I41F/C122S/A126T/V244G | 518 | 539 |
| CPC-0025366 | D23E/I41F/T98P/C122S/T144I | 519 | 540 |
| CPC-0025387 | I41F/C122S | 520 | 541 |
| CPC-0025582 | I41F/C122S/L171F/V244G | 512 | 542 |
| CPC-0025720 | C122S/H143R/Q175R | 522 | 543 |
| CPC-0025876 | I41F/C122S/L171F | 523 | 544 |
| CPC-0025974 | C122S/R230W | 524 | 545 |
| CPC-0026100 | I41F/C122S/I154V/V244G | 525 | 546 |
| CPC-0026232 | I41F/L52M/C122S/V129D/Q221(A)L | 526 | 547 |
| CPC-0027399 | F99L/C122S | 527 | 548 |
| CPC-0027706 | F97Y/C122S/I136V/Q192H/S201I | 528 | 549 |
| CPC-0027797 | H71R/C122S/P131S/D217V | 529 | 550 |
| CPC-0028017 | C122S/D217V | 530 | 551 |
| CPC-0028333 | T65K/F93L/F97Y/C 122S/D217V | 531 | 552 |
实施例12
选择的噬菌体结合的MT-SP1蛋白酶的大量制备和鉴定
A.MT-SP1噬菌体的大规模制备
本实施例描述了携带选择的MT-SP1蛋白酶结构域的噬菌体的大量制备以用于分析和随后在下游方法中使用选择的蛋白酶结构域，所述下游方法如完整MT-SP1蛋白酶的体外翻译。为此，将如以上实施例11选择的携带单噬菌体的菌落在小的无菌Corning Orange Capped Erlenmeyer培养瓶中的补加终浓度为50μg/mL的羧苄青霉素和终浓度为12μg/mL的四环素的2YT培养基中生长过夜。为制备甘油原液，将85μL 60％甘油加入到500μl的每种培养物中，随后在-80℃储存。剩余体积的每种培养物加入到含有500ml补加羧苄青霉素和四环素的2YT培养基的广口带挡板培养瓶中。或者，这一步骤可以在500mL培养瓶中以50mL体积进行。生长培养物直至OD600为0.5。
将M13KO7辅助噬菌体加入到培养物中以达到1E10CFU/ml，在37℃保温培养物1小时。加入卡那霉素至终浓度为30μg/mL。通过在37℃保温而拯救具有辅助噬菌体的培养物过夜。过夜培养后，样品在6000rpm离心15分钟。每5体积培养物加入1体积PEG/NaCl(20％PEG 8K/1.5M NaCl)溶液。然后在4℃搅拌样品20分钟。样品在10000rpm离心20分钟并除去上清。在10000rpm进行第二个离心步骤后，沉淀重悬于5mL(对于最初500mL体积)或1mL(对于最初50mL体积)PBS中。未重悬的沉淀细胞通过在14000rpm短暂离心2分钟而除去。向含有重悬的细胞的上清中以10％体积加入甘油。细胞在-80℃冷冻。
B.用ACC和QF底物测定制备的噬菌体
本实施例描述了用于评估实施例12A中制备的噬菌体的活性和特异性的荧光测定。如实施例12A所述用50mL体积培养物制备的PEG沉淀的携带突变体MT-SP1的噬菌体克隆标准化为1E13颗粒/ml。所述噬菌体然后用ACC荧光和QF(猝灭的荧光)底物如下进行酶学测定。5μL噬菌体(1E13颗粒/ml)与5μL 10X MT-SP1测定缓冲液，35μL H2O和5μL底物一起以50μL总体积加入到黑色Costar聚丙烯半孔微滴板(Corning)中。各孔中使用的底物是：Ac-SLGR-ACC(120uM终浓度)，Ac-RQAR-ACC(60uM终浓度)，或以下猝灭的荧光底物：SLGR-KI和RQAR-SA(均以0.625uM终浓度使用)。Ac-SLGR-ACC底物用于评估突变体MT-SP1克隆对靶(补体C2)裂解序列的裂解，而Ac-RQAR-ACC底物用于评估MT-SP1的天然靶裂解序列的裂解。类似地，SLGR-KI底物用于评估靶(补体C2)裂解序列的裂解，而RQAR-SA底物用于评估MT-SP1的天然靶裂解序列的裂解。这两个裂解速率的比率是选择的蛋白酶对靶定的新裂解序列的特异性的一种定量量度。选择的变体与相应的原始支架(即亲代)蛋白酶的这些比率的比较表明了选择的蛋白酶是否显示对靶定的新裂解序列的增强的选择性。对于ACC读出，SpectraMax
平板读数器被设定在380nM激发及在460nM检测发射，并具有435nM截断值。对于QF读出，SpectraMax
平板读数器被设定在490nM激发及在520nM检测发射，并具有515nM截断值。这一测定的结果见下表23。如上所述，列出了SEQ ID NO表示编码含有指明的氨基酸突变的MT-SP1蛋白酶结构域(B链)的氨基酸残基的序列以及编码具有相同突变的全长MT-SP1蛋白的氨基酸残基的序列，这些序列如以上实施例11B所述由测序确定。RFU(相对荧光单位)数字相应于对每一底物在37℃下的60分钟反应中观察到的水解速率。
表23：携带选择的MT-SP1蛋白酶结构域的噬菌体克隆的动力学测定
| 突变体MT-SP1克隆编号 | 氨基酸突变(胰凝乳蛋白酶编号) | SEQID NO.(蛋白酶结构域)： | SEQIDNO.(全长)： | Ac-SLGR-ACC(RFU/s) | Ac-RQAR-ACC(RFU/s) | SLGR-KI(RFU/s) | RQAR-SA(RFU/s) |
| 模板 | C122S | 507 | 515 | 2.4 | 23.5 | 0.12 | 0.11 |
| CPC-0028341 | I41T/C122S/P173S/Q209L | 531 | 553 | 10.9 | 56.8 | 0.36 | 0.48 |
| CPC-0033634 | F97L/C122S/F234L | 533 | 554 | 6.2 | 37.2 | 0.21 | 0.19 |
| CPC-0028971 | C122S/Q175R | 534 | 555 | 3.4 | 11.0 | 0.20 | 0.18 |
| CPC-0027484 | N95K/C122S | 535 | 556 | 2.0 | 9.5 | 0.07 | 0.05 |
| CPC-0028993 | Y60(G)S/C122S | 536 | 557 | 0.5 | 1.0 | 0.02 | 0.01 |
*RFU/s＝相对荧光单位/秒(水解速率)
实施例13
用体外反应表达选择的MT-SP1突变体蛋白
本实施例描述了如上述实施例中选择和筛选的不是基因III融合蛋白的一部分的MT-SP1蛋白酶结构域的表达。
A.将MT-SP1序列亚克隆进修饰的IVEX载体
为了表达不作为基因III融合蛋白合成的如以上实施例所述在噬菌体上选择的MT-SP1蛋白酶结构域，使用NdeI和XhoI限制位点将含有N末端激活序列和C末端6xHis标记的MT-SP1蛋白酶结构域的编码区克隆进pIVEX.2.3d RTS体外翻译载体(Roche；SEQ ID NO：559))中。在RQAR裂解位点之前的pIVEX.2.3d.MT-SP1的完整N末端氨基酸序列如SEQ ID NO：558所示：MEKTRHHHHHHSGSDCGLRSFTRQAR。使用内部SphI和BsrGI限制位点将编码MT-SP1 B链蛋白的残基亚克隆进pIVEX.2.3d.MT-SP1载体中，所述MT-SP1 B链蛋白已如上所述用噬菌粒文库选择，用实施例10B和11A所述的荧光筛选方法测定并如实施例11B所述测序。通过PCR诱变产生噬菌粒选择子(selectant)用于这一方法，所述噬菌粒选择子具有在这些内部位点之外的MT-SP1序列中的突变。
B.通过体外翻译表达MT-SP1
使用体外翻译试剂盒RTS 100大肠杆菌二硫化物试剂盒(Roche AppliedScience)表达MT-SP1蛋白酶结构域是在供应商规定的条件下采用如下优化而进行：50μl反应溶液的成分被修改为包括12μl氨基酸混合物，10μl反应混合物，12μl裂解物，并加入5μl 1M Hepes pH 8缓冲液，2.5μl 12nMTween-20，2.5μl蛋白质二硫化物异构酶(PDI)和6μl陪伴RTS GroESupplement(Roche Applied Science)。1mL进料混合物(Feeding mix)还被修改为包括168μl氨基酸混合物，24μl甲硫氨酸，608μl进料混合物，100μl1M Hepes pH 8，50μl 12nM Tween-20和50μl水。体外翻译(IVT)反应在平板摇床上于30℃保温18小时。
C.His标记的MT-SP1的纯化
在体外翻译(IVT)反应后，不溶的蛋白质经离心清除并转移到新鲜试管中。使澄清的上清(体积大约45μl)在50mM磷酸钠pH 7.5，300mM NaCl中达到1mL的终体积。将所述溶液加至在2mL多孔层析柱(Clontech)中的300μl预平衡的TALON
树脂中并使之经重力流经柱。柱用3mL含有50mM磷酸钠pH 7，300mM NaCl和7.5mM咪唑的溶液洗涤；并用600μl含有50mM磷酸钠pH 6.5，300mM NaCl和75mM咪唑的溶液洗脱。洗脱液透析进含有0.1％Tween-20磷酸缓冲盐水(PBST)中并浓缩至20μL。典型地，纯化的蛋白酶的收率是大约70％。
实施例14
突变的MT-SP1蛋白酶结构域的鉴定
本实施例描述了上述实施例13中产生的突变的突变体MT-SP1蛋白酶结构域的鉴定。
A.IVT反应的活性位点滴定
为评估蛋白酶活性，用MT-SP1抑制剂M84R Ecotin在澄清的上清中进行体外翻译的突变体MT-SP1蛋白酶结构域的活性位点滴定，如(Takeuchi et al，(1999)PNAS 96，11054-11061)所述。为此测定，IVT蛋白在1X MT-SP1活性缓冲液中被稀释至终浓度为1∶10000并与15nM Ecotin在1∶2.5系列稀释液中于30℃保温1小时。在SpectraMaxM5 MicroplateReader(Molecular Devices，Inc)上动力学评估反应对0.4mM Ac-RQAR-ACC底物的裂解。在激发波长(Ex)＝380，发射波长(Em)＝450和截断值波长(c/o)＝435检测ACC离去基团。将显示在20％至80％未抑制活性的活性分数的测定点绘制在活性对Ecotin浓度图上，并绘制一条穿过这些点的线。该线的x截距被用于建立IVT蛋白酶的活性浓度。将反应作图，其中曲线的线性部分代表IVT蛋白酶的活性浓度。因此，用活性位点滴定确定了活性位点浓度(下表24示出了若干突变体；活性位点浓度)。
B.用ACC和QF底物测定IVT MT-SP1蛋白酶结构域突变体
通过猝灭荧光(QF)动力学酶测定评估了若干IVT产生的MT-SP1噬菌体选择子对突变体RCL裂解位点相比于天然RQAR MT-SP1裂解位点的增加的特异性。如以上实施例13C所述澄清的IVT上清在1X MT-SP1活性缓冲液中1∶10000稀释并与6.25μM的天然RQAR-SL QF底物或突变体RCLC2裂解底物SLGR-KI保温。用SpectraMax
M5 Microplate Reader(Molecular Devices)经Ex波长＝490，Em波长＝520和c/o波长＝515评估裂解。IVT产生的蛋白酶对RCL靶序列相对于天然序列的相对特异性用针对SLGR-KI和RQAR-SL的RFU/s(相对荧光单位每秒)的比率计算。结果见下表24。在标示为SEQ ID NO的列中，首先列出表示含有指明的氨基酸突变的蛋白酶结构域的氨基酸序列的SEQ ID NO；表示编码含有指明的氨基酸突变的全长MT-SP1的氨基酸残基的序列的SEQ ID NO在括号中示出。RFU数字表示对于每一底物测量的相对荧光单位(水解速率)。
表24：突变体MT-SP1蛋白酶结构域的动力学测定
| 突变体MT-SP1克隆编号 | 氨基酸突变(胰凝乳蛋白酶编号) | SEQIDNO.： | 活性位点浓度 | Ac-SLGR-ACCRFU/s | Ac-RQAR-ACCRFU/s | Ac-SLGR-KIQFRFU/s | Ac-RQAR-SAQFRFU/s |
| CPC-0025720 | C122S/H143R/Q175R | 522(543) | 3.9 | 0.10 | 0.46 | 0.05 | 0.04 |
| CPC-0025876 | I41F/C122S/L171F | 523(544) | 2.5 | 0.10 | 0.39 | 0.00 | 0.01 |
| CPC-0027399 | F99L/C122S | 527(548) | 4.2 | 0.55 | 9.84 | 0.04 | 0.09 |
| 突变体MT-SP1克隆编号 | 氨基酸突变(胰凝乳蛋白酶编号) | SEQIDNO.： | 活性位点浓度 | Ac-SLGR-ACCRFU/s | Ac-RQAR-ACCRFU/s | Ac-SLGR-KIQFRFU/s | Ac-RQAR-SAQFRFU/s |
| CPC-0027797 | H71R/C122S/P131S/D217V | 529(550) | 3.7 | 0.31 | 1.62 | 0.14 | 0.19 |
| CPC-0028017 | C122S/D217V | 530(551) | 4.7 | 1.42 | 7.29 | 0.38 | 0.50 |
| CPC-0028333 | T65K/F93L/F97Y/C122S/D217V | 531(552) | 3.6 | 1.05 | 5.97 | 0.37 | 0.51 |
| 模板 | C122S | 507(515) | 3.5 | 0.22 | 3.58 | 0.05 | 0.06 |
实施例15
选择的MT-SP1突变体蛋白表达为纯化的蛋白质
A.MT-SP1蛋白酶转移进pQE载体
选择在前述实施例中测定的携带MT-SP1蛋白酶结构域的噬菌体克隆的亚集以将MT-SP1蛋白酶结构域序列转移进先前被修饰以用于表达野生型MT-SP1蛋白酶结构域的pQE30表达载体中。使用InFusion DryDownPCR克隆试剂盒(Clonetech)在供应商所规定的并如Benoit et a1.(2006)，Protein Expression & Purification 45：66-71所述的条件下将选择的克隆转移进pQE30-MT-SP1(SEQ ID NO：624)中。为此过程，用聚合酶链反应(PCR)使用pQE-Insert-F2正向引物
GTTGTTGGGGGCACGGAT(SEQ ID NO：560)和pQE-Insert-R3反向引物
ATACCCCAGTGTTCTCTTT(SEQ ID NO：561)扩增噬菌体克隆DNA编码MT-SP1蛋白酶结构域的部分，每种引物均携带非退火5’尾。不具有MT-SP1的蛋白酶结构域的质粒pQE30-MT-SP1用PCR使用正向引物pQE-Linear-F2：
TGACCGCAGCCC(SEQ ID NO：562)和反向引物pQE-Linear-R1：
GAGCTTGGACTCC(SEQ ID NO：563)线性化，随后用DpnI酶处理供体和受体PCR产物。对于上述每种线性化引物序列，18个核苷酸长的同源区即非退火5’引物尾以粗体示出。然后将受体和供体DNA混合在一起，在10μL体积中使用供应商规定的条件进行InFusion反应。将2μL反应混合物转化进50μL大肠杆菌TOP10F’感受态细胞(Invitrogen，Carlsbad，CA)。在补加100ppm羧苄青霉素的LB琼脂平板上选择菌落。从选择的克隆中分离质粒DNA并用正向引物MT-SP1-5F：GGAGAAACCGGCAGAGTAC(SEQ ID NO：564)和反向引物MT-SP1-5R：GGTTCTCGCAGGTGGTCTG(SEQ ID NO：565)测序以证实正确转移。这些引物是完全退火的。
B.突变的MT-SP1蛋白酶结构域的表达、重折叠和纯化
将实施例15B中描述的在pQE30载体(Qiagen)中编码MT-SP1变体的蛋白酶结构域的质粒转化进BL21-Gold(DE3)大肠杆菌细胞(Stratagene)中。从菌落接种含有1mL补加100μg/mL羧苄青霉素的LB的小起始培养物并在37℃保温8-10小时。用100μL这一培养物接种50mL补加100μg/mL羧苄青霉素的2xYT培养基并生长过夜。在50mL锥形试管(Corning)中，离心收获细胞然后裂解。用BugBuster
Reagent(Novagen)使用供应商提供的条件分离包涵体(IB)。IB沉淀用1mL含有100mM Tris pH 8，6M GdmHCL和20mM DTT的变性溶液溶解。在微型离心管中离心(20000×g 10分钟)除去任何不溶材料后，将上清稀释进在50mL锥形试管(Corning)中的40mL含有1.5M精氨酸，100mM Tris pH 8，150mM NaCl，5mM还原型谷胱甘肽和50μM氧化型谷胱甘肽的重折叠溶液中。试管在4℃在Nutator平台(FisherScientific)上水平放置3-4天。重折叠的尚未激活的MT-SP1变体然后在室温对25mM Tris pH 8，25mM NaCl充分透析3-4天。在透析过程中除去精氨酸后，MT-SP1蛋白酶结构域变体能够激活。
激活的MT-SP1蛋白酶结构域变体的粗制备物然后在5mL HiTrapTMQHP柱(GE Healthcare)上层析，所述的柱连接于以自动化模式运转的AKTA系统上，使得每轮能加工多达7个变体。运行缓冲液是25mM Bis-Tris pH6.5，纯化的MT-SP1在50mL梯度至350mM NaCl中洗脱。合并活性级分，然后缓冲液更换成PBS+20mM苯甲脒并用MWCO为10kDa的Amicon-Ultra 15装置(Millipore)浓缩至0.5-10mg/mL。最后，等份在液氮中闪冻并储存在-80℃。
实施例16
生物素化突变体PAI抑制剂诱饵的制备
本实施例描述了用于表达和纯化突变体PAI抑制剂蛋白的方法，所述蛋白用生物素标记以用于捕获在链霉抗生物素蛋白包被的表面上，用于从uPA文库选择变体uPA蛋白酶。根据MT-SP1变体和用于PAI中的RCL序列，这些突变体PAI抑制剂还可用于从MT-SP1文库中选择一些变体MT-SP1蛋白酶。
A.6xHis-PAI-1的N末端生物素化
为了生物素化6xHis-PAI-1或其反应中心环变体，将实施例1所述的野生型His标记的PAI-1(SEQ ID NO：625)和His标记的PAI-1变体转化进Rosetta-2(DE3)pLysS宿主菌株(Novagen)。基本上如(Blouse，G.E.，Perron，M.J.，Thompson，J.H.，Day，D.E.，Link，C.A.，and Shore，J.D.(2002)Biochemistry 41(40)，11997-12009)所述进行转化，但进行以下修改。诱导是在补加0.2％葡萄糖、100ug/mL羧苄青霉素和10ug/mL氯霉素(Cm)的2XYT培养基中于30℃进行3小时。然后如(Blouse，G.E.，Perron，M.J.，Kvassman，J.O.，Yunus，S.，Thompson，J.H.，Betts，R.L.，Lutter，L.C.，and Shore，J.D.(2003)Biochemistry 42(42)，12260-12272；Kvassman，J.-O.，and Shore，J.D.(1995)Fibrinolysis 9，215-221)所述从细胞裂解物中纯化6xHis-PAI-1的活性级分。
6xHis-PAI-1变体用二硫化物可裂解的试剂EZ-Link NHS-SS-PEO4-生物素(PIERCE，Rockford，IL #21442)在N末端优先生物素化。反应在含有50mM NaPi/300mM NaCl/1mM EDTA的缓冲液中在pH6.2、4℃下于冰上进行4小时。反应通过加入5倍摩尔过量的溶解在DMSO中的生物素化试剂而起始。反应中的DMSO的终浓度保持低于1％。通过加入0.5M Tris/1.0NaCl/10mM EDTA，pH 7.4至Tris终浓度为20mM而猝灭生物素化反应。通过对含有50mM NaPi/300mM NaCl/1mM EDTA，pH 6.2的储存缓冲液充分透析而除去过量的生物素化试剂。所得溶液中的PAI-1的浓度用0.93mLmg-1cm-1的消光系数证实(参见Kvassman，J.-O.，and Shore，J.D.(1995)Fibrinolysis 9，215-221)。生物素化程度用EZ-Quant HABA/Avidn试剂盒(PIERCE，Rockford，IL#28005)根据供应商规定的条件进行确定，典型地是每1摩尔PAI-1变体为1.0-1.2摩尔。
B.PAI的体内生物素化：BRS-TEV-OptiPAI-1stab的体内生物素化
本实施例描述了用于体内生物素化PAI的方法。在本实施例中，将合适的识别序列掺入到编码诱饵分子的基因中从而诱饵的生物素标记可以在生长的细胞中完成，而不是用纯化的诱饵在体外进行。编码具有N150H，K154T，Q319L和M354I突变的稳定的PAI-1蛋白(PAI-1stab，具有SEQ ID NO：567的氨基酸残基序列)(Berkenpas，M.B.，Lawrence，D.A.，and Ginsburg，D.(1995)EMBO J.14(13)，2969-2977)的基因被设计含有如下顺序的如下区域：1)起始密码子，2)生物素识别序列(BRS)，3)烟草蚀斑病毒蛋白酶序列(TEV)，4)PAI编码序列，5)终止密码子；用大肠杆菌密码子优化。将这一合成PAI-1stab基因用XbaI和HindIII限制酶克隆进商业表达载体pET21-a(Novagen，Madison，WI)(SEQ ID NO：566)产生质粒pCAT0002(SEQ ID NO：619)，其使用T7表达系统表达优化的PAI-1(OptiPAI-1；由SEQ ID NO：621所示氨基酸序列编码，其中氨基酸残基位置3-17和20-26分别相应于BRS和TEV位点)。质粒pCAT0002然后共转化进携带质粒pBirA(如Asai et al.，(1999)J.Biol.Chem.274：20079-20078所述)的大肠杆菌BL21-Gold(DE3)感受态细胞(Stratagene，San Diego，CA)，所述细胞过表达大肠杆菌生物素连接酶BirA。转化子在补加100ug/mL羧苄青霉素和10μg/mL氯霉素(Cm)的Luria-Bertani(LB)琼脂平板上选择。
BRS-TEV-OptiPAI-1stab(SEQ ID NO：621)及其反应中心环变体的表达基本上如(Blouse，G.E.，Perron，M.J.，Thompson，J.H.，Day，D.E.，Link，C.A.，and Shore，J.D.(2002)Biochemistry 41(40)，11997-12009)所述进行，进行如下修改。诱导通过加入0.1mM IPTG和0.1mM D-生物素在补加0.2％葡萄糖、100μg/mL羧苄青霉素和10μg/mL氯霉素(Cm)的2XYT培养基中于30℃进行3小时。BRS-TEV-OptiPAI-1stab的活性级分随后如(参见Blouse，G.E.，Perron，M.J.，Kvassman，J.O.，Yunus，S.，Thompson，J.H.，Betts，R.L.，Lutter，L.C.，and Shore，J.D.(2003)Biochemistry 42(42)，12260-12272；andKvassman，J.-O.，and Shore，J.D.(1995)Fibrinolysis 9，215-221)所述从细胞裂解物中纯化或者通过在单体抗生物素蛋白(PIERCE，Rockford，IL #20227)上根据供应商规定的条件经如下修改的层析选择而从细胞裂解物中纯化。结合缓冲液含有50mM Tris/100mM NaCl/1mM EDTA/0.01％tween-80并且pH为7.4；使用了含有这一结合缓冲液和2mM D-生物素的竞争洗脱缓冲液。来自所述竞争洗脱步骤的生物素通过对含有50mM NaPi/300mMNaCl/1mM EDTA，pH 6.2的储存缓冲液充分透析而除去。PAI-1浓度通过如(Kvassman，J.-O.，and Shore，J.D.(1995)Fibrinolysis 9，215-221)所述用0.93mL mg-1cm-1的消光系数而证实。
C.V1C OptiPAI-1stab的体外生物素化
本实施例描述了PAI变体的N末端生物素化的方法。进行本实施例中描述的方法以将合适的反应基团掺入到编码PAI诱饵分子的基因中，以便诱饵用生物素标记可以在蛋白质被纯化后完成，使得可以位置特异性标记诱饵。在本实施例中，由于天然PAI不含任何半胱氨酸残基，向编码PAI基因的DNA中加入了半胱氨酸密码子以产生含Cys的PAI，随后与Cys反应性生物素化试剂反应。
在上述质粒pCAT0002中的OptiPAI-1的N末端BRS-TEV序列通过用QuikChange-XL诱变试剂盒(Stratagene，San Diego，CA)根据供应商说明经同时导入V1C突变而缺失，产生质粒pCAT0051(SEQ ID NO：623)，其表达V1C OptiPAI-1stab蛋白(SEQ ID NO：622)。质粒pCAT0051转化进大肠杆菌BL21(DE3)pLysS感受态细胞(Stratagene，San Diego，CA)。转化体在补加100ug/mL羧苄青霉素和10ug/mL氯霉素(Cm)的Luria-Bertani(LB)琼脂平板上选择。
V1C OptiPAI-1stab蛋白及其反应中心环变体的表达基本上如(Blouse，G.E.，Perron，M.J.，Thompson，J.H.，Day，D.E.，Link，C.A.，and Shore，J.D.(2002)Biochemistry 41(40)，11997-12009)所述进行，做如下修改。诱导通过加入0.1mM IPTG在补加0.2％葡萄糖、100μg/mL羧苄青霉素和10μg/mL氯霉素的2XYT培养基中于30℃进行3小时。V1C OptiPAI-1stab或其变体的活性级分如(Blouse，G.E.，Perron，M.J.，Kvassman，J.O.，Yunus，S.，Thompson，J.H.，Betts，R.L.，Lutter，L.C.，and Shore，J.D.(2003)Biochemistry 42(42)，12260-12272；and Kvassman，J.-O.，and Shore，J.D.(1995)Fibrinolysis 9，215-221)所述从细胞裂解物中纯化。
V1C OptiPAI-1蛋白及变体用硫醇反应性可逆生物素化试剂EZ-LinkBiotin-HPDP(N-(6-(生物素酰氨基)己基)-3’-(2’-吡啶基二硫)-丙酰胺)(PIERCE，Rockford，IL #21341)在工程化的N末端半胱氨酸残基进行生物素化。根据供应商说明经如下一些修改而完成生物素缀合。生物素-HPDP的原液制备为5mg/mL于无水DMF(9.3mM)中。V1C OptiPAI-1stab在G-25凝胶过滤柱上快速脱盐，由此其被洗脱进含有50mM NaPi/150mM NaCl/1mMEDTA/0.01％Tween-80，pH 7.4的缀合缓冲液中。生物素化反应通过加入10倍摩尔过量的生物素-HPDP原液而起始。二甲基甲酰胺(DMF)的终浓度保持低于2-3％。
反应在25℃进行4小时，反应进程用在343nm嘧啶-2-硫酮离去基团的释放而跟踪。过量的生物素化试剂通过对含有50mM NaPi/300mMNaCl/1mM EDTA，pH 6.2的储存缓冲液充分透析而除去。PAI-1浓度如(Kvassman，J.-O.，and Shore，J.D.(1995)Fibrinolysis 9，215-221)所述用0.93mL mg-1cm-1的消光系数而证实。使用EZ-Quant HABA/Avidn试剂盒和嘧啶-2-硫酮(PIERCE，Rockford，IL #28005)的释放，生物素化程度典型地是每摩尔PAI-1变体为1.0-1.2摩尔生物素。
实施例17
从大肠杆菌培养物上清和周质提取物筛选MT-SP1变体
本实施例描述了两种方法，通过测定来自大肠杆菌周质空间的蛋白质或来自大肠杆菌细胞培养基的蛋白质，这两种方法各自用作筛选MT-SP1变体对噬菌体的活性的替代方法。
对于两种方法，如下制备1mL培养物。将1mL补加100ug/mL羧苄青霉素和12ug/mL四环素的2YT培养基分配到96孔深孔板的每个孔中，并用来自96孔主平板的如实施例14所述用携带MT-SP1蛋白酶结构域的噬菌体感染过夜的10μL XL-1 Blue细胞接种。深孔板用透气膜密封并置于HiGro摇床培养器中，在37℃以400rpm曝氧振荡直至细胞密度达到0.4-0.6OD600(通常振荡4-5小时)。在该点加入IPTG至终浓度为0.5mM，持续振荡生长过夜。第二天，将平板在3600rpm离心20分钟以沉淀细胞。
A.从周质制备物中筛选MT-SP1变体
使用本实施例的方法测定已被转运进大肠杆菌周质空间中的全长MT-SP1-基因III融合蛋白以及所述融合蛋白的酶活性裂解产物。在3600rpm离心后，弃去培养物上清，用以下任一条件用细胞沉淀释放周质蛋白：条件1：细胞沉淀重悬于150μL冷的磷酸缓冲盐水(PBS)中；悬浮液转移到96孔PCR板中；随后一步冻融(在-80℃20分钟/在室温水浴中10分钟)；或条件2：细胞沉淀重悬于150uL 3％BugBuster蛋白质提取试剂(Novagen)中；悬浮液转移到96孔PCR板中；以及悬浮液在室温保温30分钟。接下来，细胞悬浮液在4℃以3600rpm离心20分钟，小心取出含有周质蛋白的上清而不碰到沉淀。进一步地，周质提取物用于使用实施例10B中描述的合适底物确定MT-SP1变体的酶活性。
B.从上清制备物中筛选MT-SP1变体
使用本实施例的方法测定已经从周质扩散进入细菌细胞培养基中的全长MT-SP1-基因III融合蛋白及所述融合蛋白的催化活性碰到。在本实施例中，在离心1mL培养物后，取10μL培养物上清并用实施例10B中所述的蛋白酶测定来测定，不同的是将额外25μL测定缓冲液加入到反应中。
由于修改对于本领域技术人员是显而易见的，因此本发明仅由所附权利要求书的范围限制。
序列表
<110>催化剂生物科学公司
托里派因斯分子研究院
<120>蛋白酶筛选方法及由此鉴别的蛋白酶
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<140>Not yet assigned
<141>Herewith
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<151>2006-07-05
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Cys Leu Val Pro Val Ser Leu Ala Glu Asp Pro Gln Gly Asp Ala Ala
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Gln Lys Thr Asp Thr Ser His His Asp Gln Asp His Pro Thr Phe Asn
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Lys Ile Thr Pro Asn Leu Ala Glu Phe Ala Phe Ser Leu Tyr Arg Gln
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Ile Ala Thr Ala Phe Ala Met Leu Ser Leu Gly Thr Lys Ala Asp Thr
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His Asp Glu Ile Leu Glu Gly Leu Asn Phe Asn Leu Thr Glu Ile Pro
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Glu Ala Gln Ile His Glu Gly Phe Gln Glu Leu Leu Arg Thr Leu Asn
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Gln Pro Asp Ser Gln Leu Gln Leu Thr Thr Gly Asn Gly Leu Phe Leu
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Ser Glu Gly Leu Lys Leu Val Asp Lys Phe Leu Glu Asp Val Lys Lys
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Leu Tyr His Ser Glu Ala Phe Thr Val Asn Phe Gly Asp Thr Glu Glu
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Asn Glu Leu Thr His Asp Ile Ile Thr Lys Phe Leu Glu Asn Glu Asp
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Arg Arg Ser Ala Ser Leu His Leu Pro Lys Leu Ser Ile Thr Gly Thr
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Tyr Lys Ser Trp Leu Ile Tyr His Asn Gln His Val Leu Val Thr Pro
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Leu Ser Arg Pro Asp Thr Arg Leu Gln Leu Thr Thr Gly Ser Ser Leu
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Phe Val Asn Lys Ser Met Lys Leu Val Asp Thr Phe Leu Glu Asp Thr
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Ile Ile Arg Val Pro Met Ile Asn His Leu Gly Arg Phe Asp Ile His
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Arg Asp Arg Glu Leu Ser Ser Trp Val Leu Ala Gln His Tyr Val Gly
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Asn Ala Thr Ala Phe Phe Ile Leu Pro Asp Pro Lys Lys Met Trp Gln
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Phe Asp Ile Arg Ser Ile Asn Leu His Phe Pro Lys Leu Ser Ile Ser
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Ile Phe Ser Asn Glu Ala Asp Leu Ser Gly Val Ser Gln Glu Ala Pro
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Leu Lys Leu Ser Lys Ala Val His Val Ala Val Leu Thr Ile Asp Glu
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Arg Leu Ala Arg Ala Met Met Ala Phe Thr Ala Asp Leu Phe Ser Leu
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Val Ala Gln Thr Ser Thr Cys Pro Asn Leu Ile Leu Ser Pro Leu Ser
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Val Ala Leu Ala Leu Ser His Leu Ala Leu Gly Ala Gln Asn His Thr
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Tyr Pro Leu Arg Trp Phe Leu Leu Glu Gln Pro Glu Ile Gln Val Ala
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His Phe Pro Phe Lys Asn Asn Met Ser Phe Val Val Leu Val Pro Thr
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His Phe Glu Trp Asn Val Ser Gln Val Leu Ala Asn Leu Ser Trp Asp
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Thr Leu His Pro Pro Leu Val Trp Glu Arg Pro Thr Lys Val Arg Leu
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Arg Gly Asp Lys Leu Phe Gly Pro Asp Leu Lys Leu Val Pro Pro Met
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Ala Glu Ile His Gln Ser Phe Gln His Leu Leu Arg Thr Leu Asn Gln
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Ser Ser Asp Glu Leu Gln Leu Ser Met Gly Asn Ala Met Phe Val Lys
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Glu Gln Leu Ser Leu Leu Asp Arg Phe Thr Glu Asp Ala Lys Arg Leu
145 150 155 160
Tyr Gly Ser Glu Ala Phe Ala Thr Asp Phe Gln Asp Ser Ala Ala Ala
165 170 175
Lys Lys Leu Ile Asn Asp Tyr Val Lys Asn Gly Thr Arg Gly Lys Ile
180 185 190
Thr Asp Leu Ile Lys Asp Leu Asp Ser Gln Thr Met Met Val Leu Val
195 200 205
Asn Tyr Ile Phe Phe Lys Ala Lys Trp Glu Met Pro Phe Asp Pro Gln
210 215 220
Asp Thr His Gln Ser Arg Phe Tyr Leu Ser Lys Lys Lys Trp Val Met
225 230 235 240
Val Pro Met Met Ser Leu His His Leu Thr Ile Pro Tyr Phe Arg Asp
245 250 255
Glu Glu Leu Ser Cys Thr Val Val Glu Leu Lys Tyr Thr Gly Asn Ala
260 265 270
Ser Ala Leu Phe Ile Leu Pro Asp Gln Asp Lys Met Glu Glu Val Glu
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Ala Met Leu Leu Pro Glu Thr Leu Lys Arg Trp Arg Asp Ser Leu Glu
290 295 300
Phe Arg Glu Ile Gly Glu Leu Tyr Leu Pro Lys Phe Ser Ile Ser Arg
305 310 315 320
Asp Tyr Asn Leu Asn Asp Ile Leu Leu Gln Leu Gly Ile Glu Glu Ala
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Phe Thr Ser Lys Ala Asp Leu Ser Gly Ile Thr Gly Ala Arg Asn Leu
340 345 350
Ala Val Ser Gln Val Val His Lys Ala Val Leu Asp Val Phe Glu Glu
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Gly Thr Glu Ala Ser Ala Ala Thr Ala Val Lys Ile Thr Leu Leu Ser
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Ala Leu Val Glu Thr Arg Thr Ile Val Arg Phe Asn Arg Pro Phe Leu
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Met Ile Ile Val Pro Thr Asp Thr Gln Asn Ile Phe Phe Met Ser Lys
405 410 415
Val Thr Asn Pro Lys Gln Ala
420
<210>5
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<213>Homo sapiens
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<223>Antithrombin-III precursor SERPINC1
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Met Tyr Ser Asn Val Ile Gly Thr Val Thr Ser Gly Lys Arg Lys Val
1 5 10 15
Tyr Leu Leu Ser Leu Leu Leu Ile Gly Phe Trp Asp Cys Val Thr Cys
20 25 30
His Gly Ser Pro Val Asp Ile Cys Thr Ala Lys Pro Arg Asp Ile Pro
35 40 45
Met Asn Pro Met Cys Ile Tyr Arg Ser Pro Glu Lys Lys Ala Thr Glu
50 55 60
Asp Glu Gly Ser Glu Gln Lys Ile Pro Glu Ala Thr Asn Arg Arg Val
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Trp Glu Leu Ser Lys Ala Asn Ser Arg Phe Ala Thr Thr Phe Tyr Gln
85 90 95
His Leu Ala Asp Ser Lys Asn Asp Asn Asp Asn Ile Phe Leu Ser Pro
100 105 110
Leu Ser Ile Ser Thr Ala Phe Ala Met Thr Lys Leu Gly Ala Cys Asn
115 120 125
Asp Thr Leu Gln Gln Leu Met Glu Val Phe Lys Phe Asp Thr Ile Ser
130 135 140
Glu Lys Thr Ser Asp Gln Ile His Phe Phe Phe Ala Lys Leu Asn Cys
145 150 155 160
Arg Leu Tyr Arg Lys Ala Asn Lys Ser Ser Lys Leu Val Ser Ala Asn
165 170 175
Arg Leu Phe Gly Asp Lys Ser Leu Thr Phe Asn Glu Thr Tyr Gln Asp
180 185 190
Ile Ser Glu Leu Val Tyr Gly Ala Lys Leu Gln Pro Leu Asp Phe Lys
195 200 205
Glu Asn Ala Glu Gln Ser Arg Ala Ala Ile Asn Lys Trp Val Ser Asn
210 215 220
Lys Thr Glu Gly Arg Ile Thr Asp Val Ile Pro Ser Glu Ala Ile Asn
225 230 235 240
Glu Leu Thr Val Leu Val Leu Val Asn Thr Ile Tyr Phe Lys Gly Leu
245 250 255
Trp Lys Ser Lys Phe Ser Pro Glu Asn Thr Arg Lys Glu Leu Phe Tyr
260 265 270
Lys Ala Asp Gly Glu Ser Cys Ser Ala Ser Met Met Tyr Gln Glu Gly
275 280 285
Lys Phe Arg Tyr Arg Arg Val Ala Glu Gly Thr Gln Val Leu Glu Leu
290 295 300
Pro Phe Lys Gly Asp Asp Ile Thr Met Val Leu Ile Leu Pro Lys Pro
305 310 315 320
Glu Lys Ser Leu Ala Lys Val Glu Lys Glu Leu Thr Pro Glu Val Leu
325 330 335
Gln Glu Trp Leu Asp Glu Leu Glu Glu Met Met Leu Val Val His Met
340 345 350
Pro Arg Phe Arg Ile Glu Asp Gly Phe Ser Leu Lys Glu Gln Leu Gln
355 360 365
Asp Met Gly Leu Val Asp Leu Phe Ser Pro Glu Lys Ser Lys Leu Pro
370 375 380
Gly Ile Val Ala Glu Gly Arg Asp Asp Leu Tyr Val Ser Asp Ala Phe
385 390 395 400
His Lys Ala Phe Leu Glu Val Asn Glu Glu Gly Ser Glu Ala Ala Ala
405 410 415
Ser Thr Ala Val Val Ile Ala Gly Arg Ser Leu Asn Pro Asn Arg Val
420 425 430
Thr Phe Lys Ala Asn Arg Pro Phe Leu Val Phe Ile Arg Glu Val Pro
435 440 445
Leu Asn Thr Ile Ile Phe Met Gly Arg Val Ala Asn Pro Cys Val Lys
450 455 460
<210>6
<211>499
<212>PRT
<213>Homo sapiens
<220>
<223>Heparin cofactor 2 precursor SERPIND1
<400>6
Met Lys His Ser Leu Asn Ala Leu Leu Ile Phe Leu Ile Ile Thr Ser
1 5 10 15
Ala Trp Gly Gly Ser Lys Gly Pro Leu Asp Gln Leu Glu Lys Gly Gly
20 25 30
Glu Thr Ala Gln Ser Ala Asp Pro Gln Trp Glu Gln Leu Asn Asn Lys
35 40 45
Asn Leu Ser Met Pro Leu Leu Pro Ala Asp Phe His Lys Glu Asn Thr
50 55 60
Val Thr Asn Asp Trp Ile Pro Glu Gly Glu Glu Asp Asp Asp Tyr Leu
65 70 75 80
Asp Leu Glu Lys Ile Phe Ser Glu Asp Asp Asp Tyr Ile Asp Ile Val
85 90 95
Asp Ser Leu Ser Val Ser Pro Thr Asp Ser Asp Val Ser Ala Gly Asn
100 105 110
Ile Leu Gln Leu Phe His Gly Lys Ser Arg Ile Gln Arg Leu Asn Ile
115 120 125
Leu Asn Ala Lys Phe Ala Phe Asn Leu Tyr Arg Val Leu Lys Asp Gln
130 135 140
Val Asn Thr Phe Asp Asn Ile Phe Ile Ala Pro Val Gly Ile Ser Thr
145 150 155 160
Ala Met Gly Met Ile Ser Leu Gly Leu Lys Gly Glu Thr His Glu Gln
165 170 175
Val His Ser Ile Leu His Phe Lys Asp Phe Val Asn Ala Ser Ser Lys
180 185 190
Tyr Glu Ile Thr Thr Ile His Asn Leu Phe Arg Lys Leu Thr His Arg
195 200 205
Leu Phe Arg Arg Asn Phe Gly Tyr Thr Leu Arg Ser Val Asn Asp Leu
210 215 220
Tyr Ile Gln Lys Gln Phe Pro Ile Leu Leu Asp Phe Lys Thr Lys Val
225 230 235 240
Arg Glu Tyr Tyr Phe Ala Glu Ala Gln Ile Ala Asp Phe Ser Asp Pro
245 250 255
Ala Phe Ile Ser Lys Thr Asn Asn His Ile Met Lys Leu Thr Lys Gly
260 265 270
Leu Ile Lys Asp Ala Leu Glu Asn Ile Asp Pro Ala Thr Gln Met Met
275 280 285
Ile Leu Asn Cys Ile Tyr Phe Lys Gly Ser Trp Val Asn Lys Phe Pro
290 295 300
Val Glu Met Thr His Asn His Asn Phe Arg Leu Asn Glu Arg Glu Val
305 310 315 320
Val Lys Val Ser Met Met Gln Thr Lys Gly Asn Phe Leu Ala Ala Asn
325 330 335
Asp Gln Glu Leu Asp Cys Asp Ile Leu Gln Leu Glu Tyr Val Gly Gly
340 345 350
Ile Ser Met Leu Ile Val Val Pro His Lys Met Ser Gly Met Lys Thr
355 360 365
Leu Glu Ala Gln Leu Thr Pro Arg Val Val Glu Arg Trp Gln Lys Ser
370 375 380
Met Thr Asn Arg Thr Arg Glu Val Leu Leu Pro Lys Phe Lys Leu Glu
385 390 395 400
Lys Asn Tyr Asn Leu Val Glu Ser Leu Lys Leu Met Gly Ile Arg Met
405 410 415
Leu Phe Asp Lys Asn Gly Asn Met Ala Gly Ile Ser Asp Gln Arg Ile
420 425 430
Ala Ile Asp Leu Phe Lys His Gln Gly Thr Ile Thr Val Asn Glu Glu
435 440 445
Gly Thr Gln Ala Thr Thr Val Thr Thr Val Gly Phe Met Pro Leu Ser
450 455 460
Thr Gln Val Arg Phe Thr Val Asp Arg Pro Phe Leu Phe Leu Ile Tyr
465 470 475 480
Glu His Arg Thr Ser Cys Leu Leu Phe Met Gly Arg Val Ala Asn Pro
485 490 495
Ser Arg Ser
<210>7
<211>500
<212>PRT
<213>Homo sapiens
<220>
<223>Plasma protease C1 inhibitor precursor SERPING1
<400>7
Met Ala Ser Arg Leu Thr Leu Leu Thr Leu Leu Leu Leu Leu Leu Ala
1 5 10 15
Gly Asp Arg Ala Ser Ser Asn Pro Asn Ala Thr Ser Ser Ser Ser Gln
20 25 30
Asp Pro Glu Ser Leu Gln Asp Arg Gly Glu Gly Lys Val Ala Thr Thr
35 40 45
Val Ile Ser Lys Met Leu Phe Val Glu Pro Ile Leu Glu Val Ser Ser
50 55 60
Leu Pro Thr Thr Asn Ser Thr Thr Asn Ser Ala Thr Lys Ile Thr Ala
65 70 75 80
Asn Thr Thr Asp Glu Pro Thr Thr Gln Pro Thr Thr Glu Pro Thr Thr
85 90 95
Gln Pro Thr Ile Gln Pro Thr Gln Pro Thr Thr Gln Leu Pro Thr Asp
100 105 110
Ser Pro Thr Gln Pro Thr Thr Gly Ser Phe Cys Pro Gly Pro Val Thr
115 120 125
Leu Cys Ser Asp Leu Glu Ser His Ser Thr Glu Ala Val Leu Gly Asp
130 135 140
Ala Leu Val Asp Phe Ser Leu Lys Leu Tyr His Ala Phe Ser Ala Met
145 150 155 160
Lys Lys Val Glu Thr Asn Met Ala Phe Ser Pro Phe Ser Ile Ala Ser
165 170 175
Leu Leu Thr Gln Val Leu Leu Gly Ala Gly Glu Asn Thr Lys Thr Asn
180 185 190
Leu Glu Ser Ile Leu Ser Tyr Pro Lys Asp Phe Thr Cys Val His Gln
195 200 205
Ala Leu Lys Gly Phe Thr Thr Lys Gly Val Thr Ser Val Ser Gln Ile
210 215 220
Phe His Ser Pro Asp Leu Ala Ile Arg Asp Thr Phe Val Asn Ala Ser
225 230 235 240
Arg Thr Leu Tyr Ser Ser Ser Pro Arg Val Leu Ser Asn Asn Ser Asp
245 250 255
Ala Asn Leu Glu Leu Ile Asn Thr Trp Val Ala Lys Asn Thr Asn Asn
260 265 270
Lys Ile Ser Arg Leu Leu Asp Ser Leu Pro Ser Asp Thr Arg Leu Val
275 280 285
Leu Leu Asn Ala Ile Tyr Leu Ser Ala Lys Trp Lys Thr Thr Phe Asp
290 295 300
Pro Lys Lys Thr Arg Met Glu Pro Phe His Phe Lys Asn Ser Val Ile
305 310 315 320
Lys Val Pro Met Met Asn Ser Lys Lys Tyr Pro Val Ala His Phe Ile
325 330 335
Asp Gln Thr Leu Lys Ala Lys Val Gly Gln Leu Gln Leu Ser His Asn
340 345 350
Leu Ser Leu Val Ile Leu Val Pro Gln Asn Leu Lys His Arg Leu Glu
355 360 365
Asp Met Glu Gln Ala Leu Ser Pro Ser Val Phe Lys Ala Ile Met Glu
370 375 380
Lys Leu Glu Met Ser Lys Phe Gln Pro Thr Leu Leu Thr Leu Pro Arg
385 390 395 400
Ile Lys Val Thr Thr Ser Gln Asp Met Leu Ser Ile Met Glu Lys Leu
405 410 415
Glu Phe Phe Asp Phe Ser Tyr Asp Leu Asn Leu Cys Gly Leu Thr Glu
420 425 430
Asp Pro Asp Leu Gln Val Ser Ala Met Gln His Gln Thr Val Leu Glu
435 440 445
Leu Thr Glu Thr Gly Val Glu Ala Ala Ala Ala Ser Ala Ile Ser Val
450 455 460
Ala Arg Thr Leu Leu Val Phe Glu Val Gln Gln Pro Phe Leu Phe Val
465 470 475 480
Leu Trp Asp Gln Gln His Lys Phe Pro Val Phe Met Gly Arg Val Tyr
485 490 495
Asp Pro Arg Ala
500
<210>8
<211>406
<212>PRT
<213>Homo sapiens
<220>
<223>Plasma serine protease inhibitor precursor
SERPINA5
<400>8
Met Gln Leu Phe Leu Leu Leu Cys Leu Val Leu Leu Ser Pro Gln Gly
1 5 10 15
Ala Ser Leu His Arg His His Pro Arg Glu Met Lys Lys Arg Val Glu
20 25 30
Asp Leu His Val Gly Ala Thr Val Ala Pro Ser Ser Arg Arg Asp Phe
35 40 45
Thr Phe Asp Leu Tyr Arg Ala Leu Ala Ser Ala Ala Pro Ser Gln Asn
50 55 60
Ile Phe Phe Ser Pro Val Ser Ile Ser Met Ser Leu Ala Met Leu Ser
65 70 75 80
Leu Gly Ala Gly Ser Ser Thr Lys Met Gln Ile Leu Glu Gly Leu Gly
85 90 95
Leu Asn Leu Gln Lys Ser Ser Glu Lys Glu Leu His Arg Gly Phe Gln
100 105 110
Gln Leu Leu Gln Glu Leu Asn Gln Pro Arg Asp Gly Phe Gln Leu Ser
115 120 125
Leu Gly Asn Ala Leu Phe Thr Asp Leu Val Val Asp Leu Gln Asp Thr
130 135 140
Phe Val Ser Ala Met Lys Thr Leu Tyr Leu Ala Asp Thr Phe Pro Thr
145 150 155 160
Asn Phe Arg Asp Ser Ala Gly Ala Met Lys Gln Ile Asn Asp Tyr Val
165 170 175
Ala Lys Gln Thr Lys Gly Lys Ile Val Asp Leu Leu Lys Asn Leu Asp
180 185 190
Ser Asn Ala Val Val Ile Met Val Asn Tyr Ile Phe Phe Lys Ala Lys
195 200 205
Trp Glu Thr Ser Phe Asn His Lys Gly Thr Gln Glu Gln Asp Phe Tyr
210 215 220
Val Thr Ser Glu Thr Val Val Arg Val Pro Met Met Ser Arg Glu Asp
225 230 235 240
Gln Tyr His Tyr Leu Leu Asp Arg Asn Leu Ser Cys Arg Val Val Gly
245 250 255
Val Pro Tyr Gln Gly Asn Ala Thr Ala Leu Phe Ile Leu Pro Ser Glu
260 265 270
Gly Lys Met Gln Gln Val Glu Asn Gly Leu Ser Glu Lys Thr Leu Arg
275 280 285
Lys Trp Leu Lys Met Phe Lys Lys Arg Gln Leu Glu Leu Tyr Leu Pro
290 295 300
Lys Phe Ser Ile Glu Gly Ser Tyr Gln Leu Glu Lys Val Leu Pro Ser
305 310 315 320
Leu Gly Ile Ser Asn Val Phe Thr Ser His Ala Asp Leu Ser Gly Ile
325 330 335
Ser Asn His Ser Asn Ile Gln Val Ser Glu Met Val His Lys Ala Val
340 345 350
Val Glu Val Asp Glu Ser Gly Thr Arg Ala Ala Ala Ala Thr Gly Thr
355 360 365
Ile Phe Thr Phe Arg Ser Ala Arg Leu Asn Ser Gln Arg Leu Val Phe
370 375 380
Ash Arg Pro Phe Leu Met Phe Ile Val Asp Asn Asn Ile Leu Phe Leu
385 390 395 400
Gly Lys Val Asn Arg Pro
405
<210>9
<211>427
<212>PRT
<213>Homo sapiens
<220>
<223>Kallistatin precursor SERPINA4
<400>9
Met His Leu Ile Asp Tyr Leu Leu Leu Leu Leu Val Gly Leu Leu Ala
1 5 10 15
Leu Ser His Gly Gln Leu His Val Glu His Asp Gly Glu Ser Cys Ser
20 25 30
Asn Ser Ser His Gln Gln Ile Leu Glu Thr Gly Glu Gly Ser Pro Ser
35 40 45
Leu Lys Ile Ala Pro Ala Asn Ala Asp Phe Ala Phe Arg Phe Tyr Tyr
50 55 60
Leu Ile Ala Ser Glu Thr Pro Gly Lys Asn Ile Phe Phe Ser Pro Leu
65 70 75 80
Ser Ile Ser Ala Ala Tyr Ala Met Leu Ser Leu Gly Ala Cys Ser His
85 90 95
Ser Arg Ser Gln Ile Leu Glu Gly Leu Gly Phe Asn Leu Thr Glu Leu
100 105 110
Ser Glu Ser Asp Val His Arg Gly Phe Gln His Leu Leu His Thr Leu
115 120 125
Asn Leu Pro Gly His Gly Leu Glu Thr Arg Val Gly Ser Ala Leu Phe
130 135 140
Leu Ser His Asn Leu Lys Phe Leu Ala Lys Phe Leu Asn Asp Thr Met
145 150 155 160
Ala Val Tyr Glu Ala Lys Leu Phe His Thr Asn Phe Tyr Asp Thr Val
165 170 175
Gly Thr Ile Gln Leu Ile Asn Asp His Val Lys Lys Glu Thr Arg Gly
180 185 190
Lys Ile Val Asp Leu Val Ser Glu Leu Lys Lys Asp Val Leu Met Val
195 200 205
Leu Val Asn Tyr Ile Tyr Phe Lys Ala Leu Trp Glu Lys Pro Phe Ile
210 215 220
Ser Ser Arg Thr Thr Pro Lys Asp Phe Tyr Val Asp Glu Asn Thr Thr
225 230 235 240
Val Arg Val Pro Met Met Leu Gln Asp Gln Glu His His Trp Tyr Leu
245 250 255
His Asp Arg Tyr Leu Pro Cys Ser Val Leu Arg Met Asp Tyr Lys Gly
260 265 270
Asp Ala Thr Val Phe Phe Ile Leu Pro Asn Gln Gly Lys Met Arg Glu
275 280 285
Ile Glu Glu Val Leu Thr Pro Glu Met Leu Met Arg Trp Asn Asn Leu
290 295 300
Leu Arg Lys Arg Asn Phe Tyr Lys Lys Leu Glu Leu His Leu Pro Lys
305 310 315 320
Phe Ser Ile Ser Gly Ser Tyr Val Leu Asp Gln Ile Leu Pro Arg Leu
325 330 335
Gly Phe Thr Asp Leu Phe Ser Lys Trp Ala Asp Leu Ser Gly Ile Thr
340 345 350
Lys Gln Gln Lys Leu Glu Ala Ser Lys Ser Phe His Lys Ala Thr Leu
355 360 365
Asp Val Asp Glu Ala Gly Thr Glu Ala Ala Ala Ala Thr Ser Phe Ala
370 375 380
Ile Lys Phe Phe Ser Ala Gln Thr Asn Arg His Ile Leu Arg Phe Asn
385 390 395 400
Arg Pro Phe Leu Val Val Ile Phe Ser Thr Ser Thr Gln Ser Val Leu
405 410 415
Phe Leu Gly Lys Val Val Asp Pro Thr Lys Pro
420 425
<210>10
<211>410
<212>PRT
<213>Homo sapiens
<220>
<223>Neuroserpin precursor SERPINII
<400>10
Met Ala Phe Leu Gly Leu Phe Ser Leu Leu Val Leu Gln Ser Met Ala
1 5 10 15
Thr Gly Ala Thr Phe Pro Glu Glu Ala Ile Ala Asp Leu Ser Val Asn
20 25 30
Met Tyr Asn Arg Leu Arg Ala Thr Gly Glu Asp Glu Asn Ile Leu Phe
35 40 45
Ser Pro Leu Ser Ile Ala Leu Ala Met Gly Met Met Glu Leu Gly Ala
50 55 60
Gln Gly Ser Thr Gln Lys Glu Ile Arg His Ser Met Gly Tyr Asp Ser
65 70 75 80
Leu Lys Asn Gly Glu Glu Phe Ser Phe Leu Lys Glu Phe Ser Asn Met
85 90 95
Val Thr Ala Lys Glu Ser Gln Tyr Val Met Lys Ile Ala Asn Ser Leu
100 105 110
Phe Val Gln Asn Gly Phe His Val Asn Glu Glu Phe Leu Gln Met Met
115 120 125
Lys Lys Tyr Phe Asn Ala Ala Val Asn His Val Asp Phe Ser Gln Asn
130 135 140
Val Ala Val Ala Asn Tyr Ile Asn Lys Trp Val Glu Asn Asn Thr Asn
145 150 155 160
Asn Leu Val Lys Asp Leu Val Ser Pro Arg Asp Phe Asp Ala Ala Thr
165 170 175
Tyr Leu Ala Leu Ile Asn Ala Val Tyr Phe Lys Gly Asn Trp Lys Ser
180 185 190
Gln Phe Arg Pro Glu Asn Thr Arg Thr Phe Ser Phe Thr Lys Asp Asp
195 200 205
Glu Ser Glu Val Gln Ile Pro Met Met Tyr Gln Gln Gly Glu Phe Tyr
210 215 220
Tyr Gly Glu Phe Ser Asp Gly Ser Asn Glu Ala Gly Gly Ile Tyr Gln
225 230 235 240
Val Leu Glu Ile Pro Tyr Glu Gly Asp Glu Ile Ser Met Met Leu Val
245 250 255
Leu Ser Arg Gln Glu Val Pro Leu Ala Thr Leu Glu Pro Leu Val Lys
260 265 270
Ala Gln Leu Val Glu Glu Trp Ala Asn Ser Val Lys Lys Gln Lys Val
275 280 285
Glu Val Tyr Leu Pro Arg Phe Thr Val Glu Gln Glu Ile Asp Leu Lys
290 295 300
Asp Val Leu Lys Ala Leu Gly Ile Thr Glu Ile Phe Ile Lys Asp Ala
305 310 315 320
Asn Leu Thr Gly Leu Ser Asp Asn Lys Glu Ile Phe Leu Ser Lys Ala
325 330 335
Ile His Lys Ser Phe Leu Glu Val Asn Glu Glu Gly Ser Glu Ala Ala
340 345 350
Ala Val Ser Gly Met Ile Ala Ile Ser Arg Met Ala Val Leu Tyr Pro
355 360 365
Gln Val Ile Val Asp His Pro Phe Phe Phe Leu Ile Arg Asn Arg Arg
370 375 380
Thr Gly Thr Ile Leu Phe Met Gly Arg Val Met His Pro Glu Thr Met
385 390 395 400
Asn Thr Ser Gly His Asp Phe Glu Glu Leu
405 410
<210>11
<211>402
<212>PRT
<213>Homo sapiens
<220>
<223>Plasminogen activator inhibitor 1 precursor
SERPINE1
<400>11
Met Gln Met Ser Pro Ala Leu Thr Cys Leu Val Leu Gly Leu Ala Leu
1 5 10 15
Val Phe Gly Glu Gly Ser Ala Val His His Pro Pro Ser Tyr Val Ala
20 25 30
His Leu Ala Ser Asp Phe Gly Val Arg Val Phe Gln Gln Val AIa Gln
35 40 45
Ala Ser Lys Asp Arg Asn Val Val Phe Ser Pro Tyr Gly Val Ala Ser
50 55 60
Val Leu Ala Met Leu Gln Leu Thr Thr Gly Gly Glu Thr Gln Gln Gln
65 70 75 80
Ile Gln Ala Ala Met Gly Phe Lys Ile Asp Asp Lys Gly Met Ala Pro
85 90 95
Ala Leu Arg His Leu Tyr Lys Glu Leu Met Gly Pro Trp Asn Lys Asp
100 105 110
Glu Ile Ser Thr Thr Asp Ala Ile Phe Val Gln Arg Asp Leu Lys Leu
115 120 125
Val Gln Gly Phe Met Pro His Phe Phe Arg Leu Phe Arg Ser Thr Val
130 135 140
Lys Gln Val Asp Phe Ser Glu Val Glu Arg Ala Arg Phe Ile Ile Asn
145 150 155 160
Asp Trp Val Lys Thr His Thr Lys Gly Met Ile Ser Asn Leu Leu Gly
165 170 175
Lys Gly Ala Val Asp Gln Leu Thr Arg Leu Val Leu Val Asn Ala Leu
180 185 190
Tyr Phe Asn Gly Gln Trp Lys Thr Pro Phe Pro Asp Ser Ser Thr His
195 200 205
Arg Arg Leu Phe His Lys Ser Asp Gly Ser Thr Val Ser Val Pro Met
210 215 220
Met Ala Gln Thr Asn Lys Phe Asn Tyr Thr Glu Phe Thr Thr Pro Asp
225 230 235 240
Gly His Tyr Tyr Asp Ile Leu Glu Leu Pro Tyr His Gly Asp Thr Leu
245 250 255
Ser Met Phe Ile Ala Ala Pro Tyr Glu Lys Glu Val Pro Leu Ser Ala
260 265 270
Leu Thr Asn Ile Leu Ser Ala Gln Leu Ile Ser His Trp Lys Gly Asn
275 280 285
Met Thr Arg Leu Pro Arg Leu Leu Val Leu Pro Lys Phe Ser Leu Glu
290 295 300
Thr Glu Val Asp Leu Arg Lys Pro Leu Glu Asn Leu Gly Met Thr Asp
305 310 315 320
Met Phe Arg Gln Phe Gln Ala Asp Phe Thr Ser Leu Ser Asp Gln Glu
325 330 335
Pro Leu His Val Ala Gln Ala Leu Gln Lys Val Lys Ile Glu Val Asn
340 345 350
Glu Ser Gly Thr Val Ala Ser Ser Ser Thr Ala Val Ile Val Ser Ala
355 360 365
Arg Met Ala Pro Glu Glu Ile Ile Met Asp Arg Pro Phe Leu Phe Val
370 375 380
Val Arg His Asn Pro Thr Gly Thr Val Leu Phe Met Gly Gln Val Met
385 390 395 400
Glu Pro
<210>12
<211>405
<212>PRT
<213>Homo sapiens
<220>
<223>Myoepithelium-derived serine protease inhibitor
SERPIN12
<400>12
Met Asp Thr Ile Phe Leu Trp Ser Leu Leu Leu Leu Phe Phe Gly Ser
1 5 10 15
Gln Ala Ser Arg Cys Ser Ala Gln Lys Asn Thr Glu Phe Ala Val Asp
20 25 30
Leu Tyr Gln Glu Val Ser Leu Ser His Lys Asp Asn Ile Ile Phe Ser
35 40 45
Pro Leu Gly Ile Thr Leu Val Leu Glu Met Val Gln Leu Gly Ala Lys
50 55 60
Gly Lys Ala Gln Gln Gln Ile Arg Gln Thr Leu Lys Gln Gln Glu Thr
65 70 75 80
Ser Ala Gly Glu Glu Phe Phe Val Leu Lys Ser Phe Phe Ser Ala Ile
85 90 95
Ser Glu Lys Lys Gln Glu Phe Thr Phe Asn Leu Ala Asn Ala Leu Tyr
100 105 110
Leu Gln Glu Gly Phe Thr Val Lys Glu Gln Tyr Leu His Gly Asn Lys
115 120 125
Glu Phe Phe Gln Ser Ala Ile Lys Leu Val Asp Phe Gln Asp Ala Lys
130 135 140
Ala Cys Ala Glu Met Ile Ser Thr Trp Val Glu Arg Lys Thr Asp Gly
145 150 155 160
Lys Ile Lys Asp Met Phe Ser Gly Glu Glu Phe Gly Pro Leu Thr Arg
165 170 175
Leu Val Leu Val Asn Ala Ile Tyr Phe Lys Gly Asp Trp Lys Gln Lys
180 185 190
Phe Arg Lys Glu Asp Thr Gln Leu Ile Asn Phe Thr Lys Lys Asn Gly
195 200 205
Ser Thr Val Lys Ile Pro Met Met Lys Ala Leu Leu Arg Thr Lys Tyr
210 215 220
Gly Tyr Phe Ser Glu Ser Ser Leu Asn Tyr Gln Val Leu Glu Leu Ser
225 230 235 240
Tyr Lys Gly Asp Glu Phe Ser Leu Ile Ile Ile Leu Pro Ala Glu Gly
245 250 255
Met Asp Ile Glu Glu Val Glu Lys Leu Ile Thr Ala Gln Gln Ile Leu
260 265 270
Lys Trp Leu Ser Glu Met Gln Glu Glu Glu Val Glu Ile Ser Leu Pro
275 280 285
Arg Phe Lys Val Glu Gln Lys Val Asp Phe Lys Asp Val Leu Tyr Ser
290 295 300
Leu Asn Ile Thr Glu Ile Phe Ser Gly Gly Cys Asp Leu Ser Gly Ile
305 310 315 320
Thr Asp Ser Ser Glu Val Tyr Val Ser Gln Val Thr Gln Lys Val Phe
325 330 335
Phe Glu Ile Asn Glu Asp Gly Ser Glu Ala Ala Thr Ser Thr Gly Ile
340 345 350
His Ile Pro Val Ile Met Ser Leu Ala Gln Ser Gln Phe Ile Ala Asn
355 360 365
His Pro Phe Leu Phe Ile Met Lys His Asn Pro Thr Glu Ser Ile Leu
370 375 380
Phe Met Gly Arg Val Thr Asn Pro Asp Thr Gln Glu Ile Lys Gly Arg
385 390 395 400
Asp Leu Asp Ser Leu
405
<210>13
<211>444
<212>PRT
<213>Homo sapiens
<220>
<223>Protein Z-dependent protease inhibitor precursor
SERPINA10
<400>13
Met Lys Val Val Pro Ser Leu Leu Leu Ser Val Leu Leu Ala Gln Val
1 5 10 15
Trp Leu Val Pro Gly Leu Ala Pro Ser Pro Gln Ser Pro Glu Thr Pro
20 25 30
Ala Pro Gln Asn Gln Thr Ser Arg Val Val Gln Ala Pro Lys Glu Glu
35 40 45
Glu Glu Asp Glu Gln Glu Ala Ser Glu Glu Lys Ala Ser Glu Glu Glu
50 55 60
Lys Ala Trp Leu Met Ala Ser Arg Gln Gln Leu Ala Lys Glu Thr Ser
70 75 80
Asn Phe Gly Phe Ser Leu Leu Arg Lys Ile Ser Met Arg His Asp Gly
85 90 95
Asn Met Val Phe Ser Pro Phe Gly Met Ser Leu Ala Met Thr Gly Leu
100 105 110
Met Leu Gly Ala Thr Gly Pro Thr Glu Thr Gln Ile Lys Arg Gly Leu
115 120 125
His Leu Gln Ala Leu Lys Pro Thr Lys Pro Gly Leu Leu Pro Ser Leu
130 135 140
Phe Lys Gly Leu Arg Glu Thr Leu Ser Arg Asn Leu Glu Leu Gly Leu
145 150 155 160
Thr Gln Gly Ser Phe Ala Phe Ile His Lys Asp Phe Asp Val Lys Glu
165 170 175
Thr Phe Phe Asn Leu Ser Lys Arg Tyr Phe Asp Thr Glu Cys Val Pro
180 185 190
Met Asn Phe Arg Asn Ala Ser Gln Ala Lys Arg Leu Met Asn His Tyr
195 200 205
Ile Asn Lys Glu Thr Arg Gly Lys Ile Pro Lys Leu Phe Asp Glu Ile
210 215 220
Asn Pro Glu Thr Lys Leu Ile Leu Val Asp Tyr Ile Leu Phe Lys Gly
225 230 235 240
Lys Trp Leu Thr Pro Phe Asp Pro Val Phe Thr Glu Val Asp Thr Phe
245 250 255
His Leu Asp Lys Tyr Lys Thr Ile Lys Val Pro Met Met Tyr Gly Ala
260 265 270
Gly Lys Phe Ala Ser Thr Phe Asp Lys Asn Phe Arg Cys His Val Leu
275 280 285
Lys Leu Pro Tyr Gln Gly Asn Ala Thr Met Leu Val Val Leu Met Glu
290 295 300
Lys Met Gly Asp His Leu Ala Leu Glu Asp Tyr Leu Thr Thr Asp Leu
305 310 315 320
Val Glu Thr Trp Leu Arg Asn Met Lys Thr Arg Asn Met Glu Val Phe
325 330 335
Phe Pro Lys Phe Lys Leu Asp Gln Lys Tyr Glu Met His Glu Leu Leu
340 345 350
Arg Gln Met Gly Ile Arg Arg Ile Phe Ser Pro Phe Ala Asp Leu Ser
355 360 365
Glu Leu Ser Ala Thr Gly Arg Asn Leu Gln Val Ser Arg Val Leu Gln
370 375 380
Arg Thr Val Ile Glu Val Asp Glu Arg Gly Thr Glu Ala Val Ala Gly
385 390 395 400
Ile Leu Ser Glu Ile Thr Ala Tyr Ser Met Pro Pro Val Ile Lys Val
405 410 415
Asp Arg Pro Phe His Phe Met Ile Tyr Glu Glu Thr Ser Gly Met Leu
420 425 430
Leu Phe Leu Gly Arg Val Val Asn Pro Thr Leu Leu
435 440
<210>14
<211>398
<212>PRT
<213>Homo sapiens
<220>
<223>Glia-derived nexin precursor SERPINE2
<400>14
Met Asn Trp His Leu Pro Leu Phe Leu Leu Ala Ser Val Thr Leu Pro
1 5 10 15
Ser Ile Cys Ser His Phe Asn Pro Leu Ser Leu Glu Glu Leu Gly Ser
20 25 30
Asn Thr Gly Ile Gln Val Phe Asn Gln Ile Val Lys Ser Arg Pro His
35 40 45
Asp Asn Ile Val Ile Ser Pro His Gly Ile Ala Ser Val Leu Gly Met
50 55 60
Leu Gln Leu Gly Ala Asp Gly Arg Thr Lys Lys Gln Leu Ala Met Val
65 70 75 80
Met Arg Tyr Gly Val Asn Gly Val Gly Lys Ile Leu Lys Lys Ile Asn
85 90 95
Lys Ala Ile Val Ser Lys Lys Asn Lys Asp Ile Val Thr Val Ala Asn
100 105 110
Ala Val Phe Val Lys Asn Ala Ser Glu Ile Glu Val Pro Phe Val Thr
115 120 125
Arg Asn Lys Asp Val Phe Gln Cys Glu Val Arg Asn Val Asn Phe Glu
130 135 140
Asp Pro Ala Ser Ala Cys Asp Ser Ile Asn Ala Trp Val Lys Asn Glu
145 150 155 160
Thr Arg Asp Met Ile Asp Asn Leu Leu Ser Pro Asp Leu Ile Asp Gly
165 170 175
Val Leu Thr Arg Leu Val Leu Val Asn Ala Val Tyr Phe Lys Gly Leu
180 185 190
Trp Lys Ser Arg Phe Gln Pro Glu Asn Thr Lys Lys Arg Thr Phe Val
195 200 205
Ala Ala Asp Gly Lys Ser Tyr Gln Val Pro Met Leu Ala Gln Leu Ser
210 215 220
Val Phe Arg Cys Gly Ser Thr Ser Ala Pro Asn Asp Leu Trp Tyr Asn
225 230 235 240
Phe Ile Glu Leu Pro Tyr His Gly Glu Ser Ile Ser Met Leu Ile Ala
245 250 255
Leu Pro Thr Glu Ser Ser Thr Pro Leu Ser Ala Ile Ile Pro His Ile
260 265 270
Ser Thr Lys Thr Ile Asp Ser Trp Met Ser Ile Met Val Pro Lys Arg
275 280 285
Val Gln Val Ile Leu Pro Lys Phe Thr Ala Val Ala Gln Thr Asp Leu
290 295 300
Lys Glu Pro Leu Lys Val Leu Gly Ile Thr Asp Met Phe Asp Ser Ser
305 310 315 320
Lys Ala Asn Phe Ala Lys Ile Thr Thr Gly Ser Glu Asn Leu His Val
325 330 335
Ser His Ile Leu Gln Lys Ala Lys Ile Glu Val Ser Glu Asp Gly Thr
340 345 350
Lys Ala Ser Ala Ala Thr Thr Ala Ile Leu Ile Ala Arg Ser Ser Pro
355 360 365
Pro Trp Phe Ile Val Asp Arg Pro Phe Leu Phe Phe Ile Arg His Asn
370 375 380
Pro Thr Gly Ala Val Leu Phe Met Gly Gln Ile Asn Lys Pro
385 390 395
<210>15
<211>379
<212>PRT
<213>Homo sapiens
<220>
<223>Leukocyte elastase inhibitor SERPINB1
<400>15
Met Glu Gln Leu Ser Ser Ala Asn Thr Arg Phe Ala Leu Asp Leu Phe
1 5 10 15
Leu Ala Leu Ser Glu Asn Asn Pro Ala Gly Asn Ile Phe Ile Ser Pro
20 25 30
Phe Ser Ile Ser Ser Ala Met Ala Met Val Phe Leu Gly Thr Arg Gly
35 40 45
Asn Thr Ala Ala Gln Leu Ser Lys Thr Phe His Phe Asn Thr Val Glu
50 55 60
Glu Val His Ser Arg Phe Gln Ser Leu Asn Ala Asp Ile Asn Lys Arg
65 70 75 80
Gly Ala Ser Tyr Ile Leu Lys Leu Ala Asn Arg Leu Tyr Gly Glu Lys
85 90 95
Thr Tyr Asn Phe Leu Pro Glu Phe Leu Val Ser Thr Gln Lys Thr Tyr
100 105 110
Gly Ala Asp Leu Ala Ser Val Asp Phe Gln His Ala Ser Glu Asp Ala
115 120 125
Arg Lys Thr Ile Ash Gln Trp Val Lys Gly Gln Thr Glu Gly Lys Ile
130 135 140
Pro Glu Leu Leu Ala Ser Gly Met Val Asp Asn Met Thr Lys Leu Val
145 150 155 160
Leu Val Asn Ala Ile Tyr Phe Lys Gly Asn Trp Lys Asp Lys Phe Met
165 170 175
Lys Glu Ala Thr Thr Asn Ala Pro Phe Arg Leu Asn Lys Lys Asp Arg
180 185 190
Lys Thr Val Lys Met Met Tyr Gln Lys Lys Lys Phe Ala Tyr Gly Tyr
195 200 205
Ile Glu Asp Leu Lys Cys Arg Val Leu Glu Leu Pro Tyr Gln Gly Glu
210 215 220
Glu Leu Ser Met Val Ile Leu Leu Pro Asp Asp Ile Glu Asp Glu Ser
225 230 235 240
Thr Gly Leu Lys Lys Ile Glu Glu Gln Leu Thr Leu Glu Lys Leu His
245 250 255
Glu Trp Thr Lys Pro Glu Asn Leu Asp Phe Ile Glu Val Asn Val Ser
260 265 270
Leu Pro Arg Phe Lys Leu Glu Glu Ser Tyr Thr Leu Asn Ser Asp Leu
275 280 285
Ala Arg Leu Gly Val Gln Asp Leu Phe Asn Ser Ser Lys Ala Asp Leu
290 295 300
Ser Gly Met Ser Gly Ala Arg Asp Ile Phe Ile Ser Lys Ile Val His
305 310 315 320
Lys Ser Phe Val Glu Val Asn Glu Glu Gly Thr Glu Ala Ala Ala Ala
325 330 335
Thr Ala Gly Ile Ala Thr Phe Cys Met Leu Met Pro Glu Glu Asn Phe
340 345 350
Thr Ala Asp His Pro Phe Leu Phe Phe Ile Arg His Asn Ser Ser Gly
355 360 365
Ser Ile Leu Phe Leu Gly Arg Phe Ser Ser Pro
370 375
<210>16
<211>415
<212>PRT
<213>Homo sapiens
<220>
<223>Plasminogen activator inhibitor 2 precursor
SERPINB2
<400>16
Met Glu Asp Leu Cys Val Ala Asn Thr Leu Phe Ala Leu Asn Leu Phe
1 5 10 15
Lys His Leu Ala Lys Ala Ser Pro Thr Gln Asn Leu Phe Leu Ser Pro
20 25 30
Trp Ser Ile Ser Ser Thr Met Ala Met Val Tyr Met Gly Ser Arg Gly
35 40 45
Ser Thr Glu Asp Gln Met Ala Lys Val Leu Gln Phe Asn Glu Val Gly
50 55 60
Ala Asn Ala Val Thr Pro Met Thr Pro Glu Asn Phe Thr Ser Cys Gly
65 70 75 80
Phe Met Gln Gln Ile Gln Lys Gly Ser Tyr Pro Asp Ala Ile Leu Gln
85 90 95
Ala Gln Ala Ala Asp Lys Ile His Ser Ser Phe Arg Ser Leu Ser Ser
100 105 110
Ala Ile Asn Ala Ser Thr Gly Asn Tyr Leu Leu Glu Ser Val Asn Lys
115 120 125
Leu Phe Gly Glu Lys Ser Ala Ser Phe Arg Glu Glu Tyr Ile Arg Leu
130 135 140
Cys Gln Lys Tyr Tyr Ser Ser Glu Pro Gln Ala Val Asp Phe Leu Glu
145 150 155 160
Cys Ala Glu Glu Ala Arg Lys Lys Ile Asn Ser Trp Val Lys Thr Gln
165 170 175
Thr Lys Gly Lys Ile Pro Asn Leu Leu Pro Glu Gly Ser Val Asp Gly
180 185 190
Asp Thr Arg Met Val Leu Val Asn Ala Val Tyr Phe Lys Gly Lys Trp
195 200 205
Lys Thr Pro Phe Glu Lys Lys Leu Asn Gly Leu Tyr Pro Phe Arg Val
210 215 220
Asn Ser Ala Gln Arg Thr Pro Val Gln Met Met Tyr Leu Arg Glu Lys
225 230 235 240
Leu Asn Ile Gly Tyr Ile Glu Asp Leu Lys Ala Gln Ile Leu Glu Leu
245 250 255
Pro Tyr Ala Gly Asp Val Ser Met Phe Leu Leu Leu Pro Asp Glu Ile
260 265 270
Ala Asp Val Ser Thr Gly Leu Glu Leu Leu Glu Ser Glu Ile Thr Tyr
275 280 285
Asp Lys Leu Asn Lys Trp Thr Ser Lys Asp Lys Met Ala Glu Asp Glu
290 295 300
Val Glu Val Tyr Ile Pro Gln Phe Lys Leu Glu Glu His Tyr Glu Leu
305 310 315 320
Arg Ser Ile Leu Arg Ser Met Gly Met Glu Asp Ala Phe Asn Lys Gly
325 330 335
Arg Ala Asn Phe Ser Gly Met Ser Glu Arg Asn Asp Leu Phe Leu Ser
340 345 350
Glu Val Phe His Gln Ala Met Val Asp Val Asn Glu Glu Gly Thr Glu
355 360 365
Ala Ala Ala Gly Thr Gly Gly Val Met Thr Gly Arg Thr Gly His Gly
370 375 380
Gly Pro Gln Phe Val Ala Asp His Pro Phe Leu Phe Leu Ile Met His
385 390 395 400
Lys Ile Thr Asn Cys Ile Leu Phe Phe Gly Arg Phe Ser Ser Pro
405 410 415
<210>17
<211>376
<212>PRT
<213>Homo sapiens
<220>
<223>Placental thrombin inhibitor SERPINB6
<400>17
Met Asp Val Leu Ala Glu Ala Asn Gly Thr Phe Ala Leu Asn Leu Leu
1 5 10 15
Lys Thr Leu Gly Lys Asp Asn Ser Lys Asn Val Phe Phe Ser Pro Met
20 25 30
Ser Met Ser Cys Ala Leu Ala Met Val Tyr Met Gly Ala Lys Gly Asn
35 40 45
Thr Ala Ala Gln Met Ala Gln Ile Leu Ser Phe Asn Lys Ser Gly Gly
50 55 60
Gly Gly Asp Ile His Gln Gly Phe Gln Ser Leu Leu Thr Glu Val Asn
65 70 75 80
Lys Thr Gly Thr Gln Tyr Leu Leu Arg Val Ala Asn Arg Leu Phe Gly
85 90 95
Glu Lys Ser Cys Asp Phe Leu Ser Ser Phe Arg Asp Ser Cys Gln Lys
100 105 110
Phe Tyr Gln Ala Glu Met Glu Glu Leu Asp Phe Ile Ser Ala Val Glu
115 120 125
Lys Ser Arg Lys His Ile Asn Thr Trp Val Ala Glu Lys Thr Glu Gly
130 135 140
Lys Ile Ala Glu Leu Leu Ser Pro Gly Ser Val Asp Pro Leu Thr Arg
145 150 155 160
Leu Val Leu Val Asn Ala Val Tyr Phe Arg Gly Asn Trp Asp Glu Gln
165 170 175
Phe Asp Lys Glu Asn Thr Glu Glu Arg Leu Phe Lys Val Ser Lys Asn
180 185 190
Glu Glu Lys Pro Val Gln Met Met Phe Lys Gln Ser Thr Phe Lys Lys
195 200 205
Thr Tyr Ile Gly Glu Ile Phe Thr Gln Ile Leu Val Leu Pro Tyr Val
210 215 220
Gly Lys Glu Leu Asn Met Ile Ile Met Leu Pro Asp Glu Thr Thr Asp
225 230 235 240
Leu Arg Thr Val Glu Lys Glu Leu Thr Tyr Glu Lys Phe Val Glu Trp
245 250 255
Thr Arg Leu Asp Met Met Asp Glu Glu Glu Val Glu Val Ser Leu Pro
260 265 270
Arg Phe Lys Leu Glu Glu Ser Tyr Asp Met Glu Ser Val Leu Arg Asn
275 280 285
Leu Gly Met Thr Asp Ala Phe Glu Leu Gly Lys Ala Asp Phe Ser Gly
290 295 300
Met Ser Gln Thr Asp Leu Ser Leu Ser Lys Val Val His Lys Ser Phe
305 310 315 320
Val Glu Val Asn Glu Glu Gly Thr Glu Ala Ala Ala Ala Thr Ala Ala
325 330 335
Ile Met Met Met Arg Cys Ala Arg Phe Val Pro Arg Phe Cys Ala Asp
340 345 350
His Pro Phe Leu Phe Phe Ile Gln His Ser Lys Thr Asn Gly Ile Leu
355 360 365
Phe Cys Gly Arg Phe Ser Ser Pro
370 375
<210>18
<211>397
<212>PRT
<213>Homo sapiens
<220>
<223>Bomapin SERPINB10
<400>18
Met Asp Ser Leu Ala Thr Ser Ile Asn Gln Phe Ala Leu Glu Leu Ser
1 5 10 15
Lys Lys Leu Ala Glu Ser Ala Gln Gly Lys Asn Ile Phe Phe Ser Ser
20 25 30
Trp Ser Ile Ser Thr Ser Leu Thr Ile Val Tyr Leu Gly Ala Lys Gly
35 40 45
Thr Thr Ala Ala Gln Met Ala Gln Val Leu Gln Phe Asn Arg Asp Gln
50 55 60
Gly Val Lys Cys Asp Pro Glu Ser Glu Lys Lys Arg Lys Met Glu Phe
65 70 75 80
Asn Leu Ser Asn Ser Glu Glu Ile His Ser Asp Phe Gln Thr Leu Ile
85 90 95
Ser Glu Ile Leu Lys Pro Asn Asp Asp Tyr Leu Leu Lys Thr Ala Asn
100 105 110
Ala Ile Tyr Gly Glu Lys Thr Tyr Ala Phe His Asn Lys Tyr Leu Glu
115 120 125
Asp Met Lys Thr Tyr Phe Gly Ala Glu Pro Gln Pro Val Asn Phe Val
130 135 140
Glu Ala Ser Asp Gln Ile Arg Lys Asp Ile Asn Ser Trp Val Glu Arg
145 150 155 160
Gln Thr Glu Gly Lys Ile Gln Asn Leu Leu Pro Asp Asp Ser Val Asp
165 170 175
Ser Thr Thr Arg Met Ile Leu Val Asn Ala Leu Tyr Phe Lys Gly Ile
180 185 190
Trp Glu His Gln Phe Leu Val Gln Asn Thr Thr Glu Lys Pro Phe Arg
195 200 205
Ile Asn Glu Thr Thr Ser Lys Pro Val Gln Met Met Phe Met Lys Lys
210 215 220
Lys Leu His Ile Phe His Ile Glu Lys Pro Lys Ala Val Gly Leu Gln
225 230 235 240
Leu Tyr Tyr Lys Ser Arg Asp Leu Ser Leu Leu Ile Leu Leu Pro Glu
245 250 255
Asp Ile Asn Gly Leu Glu Gln Leu Glu Lys Ala Ile Thr Tyr Glu Lys
260 265 270
Leu Asn Glu Trp Thr Ser Ala Asp Met Met Glu Leu Tyr Glu Val Gln
275 280 285
Leu His Leu Pro Lys Phe Lys Leu Glu Asp Ser Tyr Asp Leu Lys Ser
290 295 300
Thr Leu Ser Ser Met Gly Met Ser Asp Ala Phe Ser Gln Ser Lys Ala
305 310 315 320
Asp Phe Ser Gly Met Ser Ser Ala Arg Asn Leu Phe Leu Ser Asn Val
325 330 335
Phe His Lys Ala Phe Val Glu Ile Asn Glu Gln Gly Thr Glu Ala Ala
340 345 350
Ala Gly Ser Gly Ser Glu Ile Asp Ile Arg Ile Arg Val Pro Ser Ile
355 360 365
Glu Phe Asn Ala Asn His Pro Phe Leu Phe Phe Ile Arg His Asn Lys
370 375 380
Thr Asn Thr Ile Leu Phe Tyr Gly Arg Leu Cys Ser Pro
385 390 395
<210>19
<211>392
<212>PRT
<213>Homo sapiens
<220>
<223>Epipen SERPINB11
<400>19
Met Gly Ser Leu Ser Thr Ala Asn Val Glu Phe Cys Leu Asp Val Phe
1 5 10 15
Lys Glu Leu Asn Ser Asn Asn Ile Gly Asp Asn Ile Phe Phe Ser Ser
20 25 30
Leu Ser Leu Leu Tyr Ala Leu Ser Met Val Leu Leu Gly Ala Arg Gly
35 40 45
Glu Thr Ala Glu Gln Leu Glu Lys Val Leu His Phe Ser His Thr Val
50 55 60
Asp Ser Leu Lys Pro Gly Phe Lys Asp Ser Pro Lys Cys Ser Gln Ala
65 70 75 80
Gly Arg Ile His Ser Glu Phe Gly Val Glu Phe Ser Gln Ile Asn Gln
85 90 95
Pro Asp Ser Asn Cys Thr Leu Ser Ile Ala Asn Arg Leu Tyr Gly Thr
100 105 110
Lys Thr Met Ala Phe His Gln Gln Tyr Leu Ser Cys Ser Glu Lys Trp
115 120 125
Tyr Gln Ala Arg Leu Gln Thr Val Asp Phe Glu Gln Ser Thr Glu Glu
130 135 140
Thr Arg Lys Met Ile Asn Ala Trp Val Glu Asn Lys Thr Asn Gly Lys
145 150 155 160
Val Ala Asn Leu Phe Gly Lys Ser Thr Ile Asp Pro Ser Ser Val Met
165 170 175
Val Leu Val Asn Ile Ile Tyr Phe Lys Gly Gln Arg Gln Asn Lys Phe
180 185 190
Gln Val Arg Glu Thr Val Lys Ser Pro Phe Gln Leu Ser Glu Gly Lys
195 200 205
Asn Val Thr Val Glu Met Met Tyr Gln Ile Gly Thr Phe Lys Leu Ala
210 215 220
Phe Val Lys Glu Pro Gln Met Gln Val Leu Glu Leu Pro Tyr Val Asn
225 230 235 240
Asn Lys Leu Ser Met Ile Ile Leu Leu Pro Val Gly Ile Ala Asn Leu
245 250 255
Lys Gln Ile Glu Lys Gln Leu Asn Ser Gly Thr Phe His Glu Trp Thr
260 265 270
Ser Ser Ser Asn Met Met Glu Arg Glu Val Glu Val His Leu Pro Arg
275 280 285
Phe Lys Leu Glu Ile Lys Tyr Glu Leu Asn Ser Leu Leu Lys Pro Leu
290 295 300
Gly Val Thr Asp Leu Phe Asn Gln Val Lys Ala Asp Leu Ser Gly Met
305 310 315 320
Ser Pro Thr Lys Gly Leu Tyr Leu Ser Lys Ala Ile His Lys Ser Tyr
325 330 335
Leu Asp Val Ser Glu Glu Gly Thr Glu Ala Ala Ala Ala Thr Gly Asp
340 345 350
Ser Ile Ala Val Lys Ser Leu Pro Met Arg Ala Gln Phe Lys Ala Asn
355 360 365
His Pro Phe Leu Phe Phe Ile Arg His Thr His Thr Asn Thr Ile Leu
370 375 380
Phe Cys Gly Lys Leu Ala Ser Pro
385 390
<210>20
<211>405
<212>PRT
<213>Homo sapiens
<220>
<223>Yukopin SERPINB12
<400>20
Met Asp Ser Leu Val Thr Ala Asn Thr Lys Phe Cys Phe Asp Leu Phe
1 5 10 15
Gln Glu Ile Gly Lys Asp Asp Arg His Lys Asn Ile Phe Phe Ser Pro
20 25 30
Leu Ser Leu Ser Ala Ala Leu Gly Met Val Arg Leu Gly Ala Arg Ser
35 40 45
Asp Ser Ala His Gln Ile Asp Glu Val Leu His Phe Asn Glu Phe Ser
50 55 60
Gln Asn Glu Ser Lys Glu Pro Asp Pro Cys Leu Lys Ser Asn Lys Gln
65 70 75 80
Lys Ala Gly Ser Leu Asn Asn Glu Ser Gly Leu Val Ser Cys Tyr Phe
85 90 95
Gly Gln Leu Leu Ser Lys Leu Asp Arg Ile Lys Thr Asp Tyr Thr Leu
100 105 110
Ser Ile Ala Asn Arg Leu Tyr Gly Glu Gln Glu Phe Pro Ile Cys Gln
115 120 125
Glu Tyr Leu Asp Gly Val Ile Gln Phe Tyr His Thr Thr Ile Glu Ser
130 135 140
Val Asp Phe Gln Lys Asn Pro Glu Lys Ser Arg Gln Glu Ile Asn Phe
145 150 155 160
Trp Val Glu Cys Gln Ser Gln Gly Lys Ile Lys Glu Leu Phe Ser Lys
165 170 175
Asp Ala Ile Asn Ala Glu Thr Val Leu Val Leu Val Asn Ala Val Tyr
180 185 190
Phe Lys Ala Lys Trp Glu Thr Tyr Phe Asp His Glu Asn Thr Val Asp
195 200 205
Ala Pro Phe Cys Leu Asn Ala Asn Glu Asn Lys Ser Val Lys Met Met
210 215 220
Thr Gln Lys Gly Leu Tyr Arg Ile Gly Phe Ile Glu Glu Val Lys Ala
225 230 235 240
Gln Ile Leu Glu Met Arg Tyr Thr Lys Gly Lys Leu Ser Met Phe Val
245 250 255
Leu Leu Pro Ser His Ser Lys Asp Asn Leu Lys Gly Leu Glu Glu Leu
260 265 270
Glu Arg Lys Ile Thr Tyr Glu Lys Met Val Ala Trp Ser Ser Ser Glu
275 280 285
Asn Met Ser Glu Glu Ser Val Val Leu Ser Phe Pro Arg Phe Thr Leu
290 295 300
Glu Asp Ser Tyr Asp Leu Asn Ser Ile Leu Gln Asp Met Gly Ile Thr
305 310 315 320
Asp Ile Phe Asp Glu Thr Arg Ala Asp Leu Thr Gly Ile Ser Pro Ser
325 330 335
Pro Asn Leu Tyr Leu Ser Lys Ile Ile His Lys Thr Phe Val Glu Val
340 345 350
Asp Glu Asn Gly Thr Gln Ala Ala Ala Ala Thr Gly Ala Val Val Ser
355 360 365
Glu Arg Ser Leu Arg Ser Trp Val Glu Phe Asn Ala Asn His Pro Phe
370 375 380
Leu Phe Phe Ile Arg His Asn Lys Thr Gln Thr Ile Leu Phe Tyr Gly
385 390 395 400
Arg Val Cys Ser Pro
405
<210>21
<211>391
<212>PRT
<213>Homo sapiens
<220>
<223>Headpin SERPINB13
<400>21
Met Asp Ser Leu Gly Ala Val Ser Thr Arg Leu Gly Phe Asp Leu Phe
1 5 10 15
Lys Glu Leu Lys Lys Thr Asn Asp Gly Asn Ile Phe Phe Ser Pro Val
20 25 30
Gly Ile Leu Thr Ala Ile Gly Met Val Leu Leu Gly Thr Arg Gly Ala
35 40 45
Thr Ala Ser Gln Leu Glu Glu Val Phe His Ser Glu Lys Glu Thr Lys
50 55 60
Ser Ser Arg Ile Lys Ala Glu Glu Lys Glu Val Ile Glu Asn Thr Glu
65 70 75 80
Ala Val His Gln Gln Phe Gln Lys Phe Leu Thr Glu Ile Ser Lys Leu
85 90 95
Thr Asn Asp Tyr Glu Leu Asn Ile Thr Asn Arg Leu Phe Gly Glu Lys
100 105 110
Thr Tyr Leu Phe Leu Gln Lys Tyr Leu Asp Tyr Val Glu Lys Tyr Tyr
115 120 125
His Ala Ser Leu Glu Pro Val Asp Phe Val Asn Ala Ala Asp Glu Ser
130 135 140
Arg Lys Lys Ile Asn Ser Trp Val Glu Ser Lys Thr Asn Glu Lys Ile
145 150 155 160
Lys Asp Leu Phe Pro Asp Gly Ser Ile Ser Ser Ser Thr Lys Leu Val
165 170 175
Leu Val Asn Met Val Tyr Phe Lys Gly Gln Trp Asp Arg Glu Phe Lys
180 185 190
Lys Glu Asn Thr Lys Glu Glu Lys Phe Trp Met Asn Lys Ser Thr Ser
195 200 205
Lys Ser Val Gln Met Met Thr Gln Ser His Ser Phe Ser Phe Thr Phe
210 215 220
Leu Glu Asp Leu Gln Ala Lys Ile Leu Gly Ile Pro Tyr Lys Asn Asn
225 230 235 240
Asp Leu Ser Met Phe Val Leu Leu Pro Asn Asp Ile Asp Gly Leu Glu
245 250 255
Lys Ile Ile Asp Lys Ile Ser Pro Glu Lys Leu Val Glu Trp Thr Ser
260 265 270
Pro Gly His Met Glu Glu Arg Lys Val Asn Leu His Leu Pro Arg Phe
275 280 285
Glu Val Glu Asp Gly Tyr Asp Leu Glu Ala Val Leu Ala Ala Met Gly
290 295 300
Met Gly Asp Ala Phe Ser Glu His Lys Ala Asp Tyr Ser Gly Met Ser
305 310 315 320
Ser Gly Ser Gly Leu Tyr Ala Gln Lys Phe Leu His Ser Ser Phe Val
325 330 335
Ala Val Thr Glu Glu Gly Thr Glu Ala Ala Ala Ala Thr Gly Ile Gly
340 345 350
Phe Thr Val Thr Ser Ala Pro Gly His Glu Asn Val His Cys Asn His
355 360 365
Pro Phe Leu Phe Phe Ile Arg His Asn Glu Ser Asn Ser Ile Leu Phe
370 375 380
Phe Gly Arg Phe Ser Ser Pro
385 390
<210>22
<211>390
<212>PRT
<213>Homo sapiens
<220>
<223>Squamous cell carcinoma antigen 1 SERPINB3
<400>22
Met Asn Ser Leu Ser Glu Ala Asn Thr Lys Phe Met Phe Asp Leu Phe
1 5 10 15
Gln Gln Phe Arg Lys Ser Lys Glu Asn Asn Ile Phe Tyr Ser Pro Ile
20 25 30
Ser Ile Thr Ser Ala Leu Gly Met Val Leu Leu Gly Ala Lys Asp Asn
35 40 45
Thr Ala Gln Gln Ile Lys Lys Val Leu His Phe Asp Gln Val Thr Glu
50 55 60
Asn Thr Thr Gly Lys Ala Ala Thr Tyr His Val Asp Arg Ser Gly Asn
65 70 75 80
Val His His Gln Phe Gln Lys Leu Leu Thr Glu Phe Asn Lys Ser Thr
85 90 95
Asp Ala Tyr Glu Leu Lys Ile Ala Asn Lys Leu Phe Gly Glu Lys Thr
100 105 110
Tyr Leu Phe Leu Gln Glu Tyr Leu Asp Ala Ile Lys Lys Phe Tyr Gln
115 120 125
Thr Ser Val Glu Ser Val Asp Phe Ala Asn Ala Pro Glu Glu Ser Arg
130 135 140
Lys Lys Ile Asn Ser Trp Val Glu Ser Gln Thr Asn Glu Lys Ile Lys
145 150 155 160
Asn Leu Ile Pro Glu Gly Asn Ile Gly Ser Asn Thr Thr Leu Val Leu
165 170 175
Val Asn Ala Ile Tyr Phe Lys Gly Gln Trp Glu Lys Lys Phe Asn Lys
180 185 190
Glu Asp Thr Lys Glu Glu Lys Phe Trp Pro Asn Lys Asn Thr Tyr Lys
195 200 205
Ser Ile Gln Met Met Arg Gln Tyr Thr Ser Phe His Phe Ala Ser Leu
210 215 220
Glu Asp Val Gln Ala Lys Val Leu Glu Ile Pro Tyr Lys Gly Lys Asp
225 230 235 240
Leu Ser Met Ile Val Leu Leu Pro Asn Glu Ile Asp Gly Leu Gln Lys
245 250 255
Leu Glu Glu Lys Leu Thr Ala Glu Lys Leu Met Glu Trp Thr Ser Leu
260 265 270
Gln Asn Met Arg Glu Thr Arg Val Asp Leu His Leu Pro Arg Phe Lys
275 280 285
Val Glu Glu Ser Tyr Asp Leu Lys Asp Thr Leu Arg Thr Met Gly Met
290 295 300
Val Asp Ile Phe Asn Gly Asp Ala Asp Leu Ser Gly Met Thr Gly Ser
305 310 315 320
Arg Gly Leu Val Leu Ser Gly Val Leu His Lys Ala Phe Val Glu Val
325 330 335
Thr Glu Glu Gly Ala Glu Ala Ala Ala Ala Thr Ala Val Val Gly Phe
340 345 350
Gly Ser Ser Pro Thr Ser Thr Asn Glu Glu Phe His Cys Asn His Pro
355 360 365
Phe Leu Phe Phe Ile Arg Gln Asn Lys Thr Asn Ser Ile Leu Phe Tyr
370 375 380
Gly Arg Phe Ser Ser Pro
385 390
<210>23
<211>390
<212>PRT
<213>Homo sapiens
<220>
<223>Squamous cell carcinoma antigen 2 SERPINB4
<400>23
Met Asn Ser Leu Ser Glu Ala Asn Thr Lys Phe Met Phe Asp Leu Phe
1 5 10 15
Gln Gln Phe Arg Lys Ser Lys Glu Asn Asn Ile Phe Tyr Ser Pro Ile
20 25 30
Ser Ile Thr Ser Ala Leu Gly Met Val Leu Leu Gly Ala Lys Asp Asn
35 40 45
Thr Ala Gln Gln Ile Ser Lys Val Leu His Phe Asp Gln Val Thr Glu
50 55 60
Asn Thr Thr Glu Lys Ala Ala Thr Tyr His Val Asp Arg Ser Gly Asn
65 70 75 80
Val His His Gln Phe Gln Lys Leu Leu Thr Glu Phe Asn Lys Ser Thr
85 90 95
Asp Ala Tyr Glu Leu Lys Ile Ala Asn Lys Leu Phe Gly Glu Lys Thr
100 105 110
Tyr Gln Phe Leu Gln Glu Tyr Leu Asp Ala Ile Lys Lys Phe Tyr Gln
115 120 125
Thr Ser Val Glu Ser Thr Asp Phe Ala Asn Ala Pro Glu Glu Ser Arg
130 135 140
Lys Lys Ile Asn Ser Trp Val Glu Ser Gln Thr Asn Glu Lys Ile Lys
145 150 155 160
Asn Leu Phe Pro Asp Gly Thr Ile Gly Asn Asp Thr Thr Leu Val Leu
165 170 175
Val Asn Ala Ile Tyr Phe Lys Gly Gln Trp Glu Asn Lys Phe Lys Lys
180 185 190
Glu Asn Thr Lys Glu Glu Lys Phe Trp Pro Asn Lys Asn Thr Tyr Lys
195 200 205
Ser Val Gln Met Met Arg Gln Tyr Asn Ser Phe Asn Phe Ala Leu Leu
210 215 220
Glu Asp Val Gln Ala Lys Val Leu Glu Ile Pro Tyr Lys Gly Lys Asp
225 230 235 240
Leu Ser Met Ile Val Leu Leu Pro Asn Glu Ile Asp Gly Leu Gln Lys
245 250 255
Leu Glu Glu Lys Leu Thr Ala Glu Lys Leu Met Glu Trp Thr Ser Leu
260 265 270
Gln Asn Met Arg Glu Thr Cys Val Asp Leu His Leu Pro Arg Phe Lys
275 280 285
Met Glu Glu Ser Tyr Asp Leu Lys Asp Thr Leu Arg Thr Met Gly Met
290 295 300
Val Asn Ile Phe Asn Gly Asp Ala Asp Leu Ser Gly Met Thr Trp Ser
305 310 315 320
His Gly Leu Ser Val Ser Lys Val Leu His Lys Ala Phe Val Glu Val
325 330 335
Thr Glu Glu Gly Val Glu Ala Ala Ala Ala Thr Ala Val Val Val Val
340 345 350
Glu Leu Ser Ser Pro Ser Thr Asn Glu Glu Phe Cys Cys Asn His Pro
355 360 365
Phe Leu Phe Phe Ile Arg Gln Asn Lys Thr Asn Ser Ile Leu Phe Tyr
370 375 380
Gly Arg Phe Ser Ser Pro
385 390
<210>24
<211>380
<212>PRT
<213>Homo sapiens
<220>
<223>Megsin SERPINB7
<400>24
Met Ala Ser Leu Ala Ala Ala Asn Ala Glu Phe Cys Phe Asn Leu Phe
1 5 10 15
Arg Glu Met Asp Asp Asn Gln Gly Asn Gly Asn Val Phe Phe Ser Ser
20 25 30
Leu Ser Leu Phe Ala Ala Leu Ala Leu Val Arg Leu Gly Ala Gln Asp
35 40 45
Asp Ser Leu Ser Gln Ile Asp Lys Leu Leu His Val Asn Thr Ala Ser
50 55 60
Gly Tyr Gly Asn Ser Ser Asn Ser Gln Ser Gly Leu Gln Ser Gln Leu
65 70 75 80
Lys Arg Val Phe Ser Asp Ile Asn Ala Ser His Lys Asp Tyr Asp Leu
85 90 95
Ser Ile Val Asn Gly Leu Phe Ala Glu Lys Val Tyr Gly Phe His Lys
100 105 110
Asp Tyr Ile Glu Cys Ala Glu Lys Leu Tyr Asp Ala Lys Val Glu Arg
115 120 125
Val Asp Phe Thr Asn His Leu Glu Asp Thr Arg Arg Asn Ile Asn Lys
130 135 140
Trp Val Glu Asn Glu Thr His Gly Lys Ile Lys Asn Val Ile Gly Glu
145 150 155 160
Gly Gly Ile Ser Ser Ser Ala Val Met Val Leu Val Asn Ala Val Tyr
165 170 175
Phe Lys Gly Lys Trp Gln Ser Ala Phe Thr Lys Ser Glu Thr Ile Asn
180 185 190
Cys His Phe Lys Ser Pro Lys Cys Ser Gly Lys Ala Val Ala Met Met
195 200 205
His Gln Glu Arg Lys Phe Asn Leu Ser Val Ile Glu Asp Pro Ser Met
210 215 220
Lys Ile Leu Glu Leu Arg Tyr Asn Gly Gly Ile Asn Met Tyr Val Leu
225 230 235 240
Leu Pro Glu Asn Asp Leu Ser Glu Ile Glu Asn Lys Leu Thr Phe Gln
245 250 255
Asn Leu Met Glu Trp Thr Asn Pro Arg Arg Met Thr Ser Lys Tyr Val
260 265 270
Glu Val Phe Phe Pro Gln Phe Lys Ile Glu Lys Asn Tyr Glu Met Lys
275 280 285
Gln Tyr Leu Arg Ala Leu Gly Leu Lys Asp Ile Phe Asp Glu Ser Lys
290 295 300
Ala Asp Leu Ser Gly Ile Ala Ser Gly Gly Arg Leu Tyr Ile Ser Arg
305 310 315 320
Met Met His Lys Ser Tyr Ile Glu Val Thr Glu Glu Gly Thr Glu Ala
325 330 335
Thr Ala Ala Thr Gly Ser Asn Ile Val Glu Lys Gln Leu Pro Gln Ser
340 345 350
Thr Leu Phe Arg Ala Asp His Pro Phe Leu Phe Val Ile Arg Lys Asp
355 360 365
Asp Ile Ile Leu Phe Ser Gly Lys Val Ser Cys Pro
370 375 380
<210>25
<211>374
<212>PRT
<213>Homo sapiens
<220>
<223>Cytoplasmic antiproteinase 2 SERPINB8
<400>25
Met Asp Asp Leu Cys Glu Ala Asn Gly Thr Phe Ala Ile Ser Leu Phe
1 5 10 15
Lys Ile Leu Gly Glu Glu Asp Asn Ser Arg Asn Val Phe Phe Ser Pro
20 25 30
Met Ser Ile Ser Ser Ala Leu Ala Met Val Phe Met Gly Ala Lys Gly
35 40 45
Ser Thr Ala Ala Gln Met Ser Gln Ala Leu Cys Leu Tyr Lys Asp Gly
50 55 60
Asp Ile His Arg Gly Phe Gln Ser Leu Leu Ser Glu Val Asn Arg Thr
65 70 75 80
Gly Thr Gln Tyr Leu Leu Arg Thr Ala Asn Arg Leu Phe Gly Glu Lys
85 90 95
Thr Cys Asp Phe Leu Pro Asp Phe Lys Glu Tyr Cys Gln Lys Phe Tyr
100 105 110
Gln Ala Glu Leu Glu Glu Leu Ser Phe Ala Glu Asp Thr Glu Glu Cys
115 120 125
Arg Lys His Ile Asn Asp Trp Val Ala Glu Lys Thr Glu Gly Lys Ile
130 135 140
Ser Glu Val Leu Asp Ala Gly Thr Val Asp Pro Leu Thr Lys Leu Val
145 150 155 160
Leu Val Asn Ala Ile Tyr Phe Lys Gly Lys Trp Asn Glu Gln Phe Asp
165 170 175
Arg Lys Tyr Thr Arg Gly Met Leu Phe Lys Thr Asn Glu Glu Lys Lys
180 185 190
Thr Val Gln Met Met Phe Lys Glu Ala Lys Phe Lys Met Gly Tyr Ala
195 200 205
Asp Glu Val His Thr Gln Val Leu Glu Leu Pro Tyr Val Glu Glu Glu
210 215 220
Leu Ser Met Val Ile Leu Leu Pro Asp Asp Asn Thr Asp Leu Ala Val
225 230 235 240
Val Glu Lys Ala Leu Thr Tyr Glu Lys Phe Lys Ala Trp Thr Asn Ser
245 250 255
Glu Lys Leu Thr Lys Ser Lys Val Gln Val Phe Leu Pro Arg Leu Lys
260 265 270
Leu Glu Glu Ser Tyr Asp Leu Glu Pro Phe Leu Arg Arg Leu Gly Met
275 280 285
Ile Asp Ala Phe Asp Glu Ala Lys Ala Asp Phe Ser Gly Met Ser Thr
290 295 300
Glu Lys Asn Val Pro Leu Ser Lys Val Ala His Lys Cys Phe Val Glu
305 310 315 320
Val Asn Glu Glu Gly Thr Glu Ala Ala Ala Ala Thr Ala Val Val Arg
325 330 335
Asn Ser Arg Cys Ser Arg Met Glu Pro Arg Phe Cys Ala Asp His Pro
340 345 350
Phe Leu Phe Phe Ile Arg Arg His Lys Thr Asn Cys Ile Leu Phe Cys
355 360 365
Gly Arg Phe Ser Ser Pro
370
<210>26
<211>376
<212>PRT
<213>Homo sapiens
<220>
<223>Cytoplasmic antiproteinase 3 SERPINB9
<400>26
Met Glu Thr Leu Ser Asn Ala Ser Gly Thr Phe Ala Ile Arg Leu Leu
1 5 10 15
Lys Ile Leu Cys Gln Asp Asn Pro Ser His Asn Val Phe Cys Ser Pro
20 25 30
Val Ser Ile Ser Ser Ala Leu Ala Met Val Leu Leu Gly Ala Lys Gly
35 40 45
Asn Thr Ala Thr Gln Met Ala Gln Ala Leu Ser Leu Asn Thr Glu Glu
50 55 60
Asp Ile His Arg Ala Phe Gln Ser Leu Leu Thr Glu Val Asn Lys Ala
65 70 75 80
Gly Thr Gln Tyr Leu Leu Arg Thr Ala Asn Arg Leu Phe Gly Glu Lys
85 90 95
Thr Cys Gln Phe Leu Ser Thr Phe Lys Glu Ser Cys Leu Gln Phe Tyr
100 105 110
His Ala Glu Leu Lys Glu Leu Ser Phe Ile Arg Ala Ala Glu Glu Ser
115 120 125
Arg Lys His Ile Asn Thr Trp Val Ser Lys Lys Thr Glu Gly Lys Ile
130 135 140
Glu Glu Leu Leu Pro Gly Ser Ser Ile Asp Ala Glu Thr Arg Leu Val
145 150 155 160
Leu Val Asn Ala Ile Tyr Phe Lys Gly Lys Trp Asn Glu Pro Phe Asp
165 170 175
Glu Thr Tyr Thr Arg Glu Met Pro Phe Lys Ile Asn Gln Glu Glu Gln
180 185 190
Arg Pro Val Gln Met Met Tyr Gln Glu Ala Thr Phe Lys Leu Ala His
195 200 205
Val Gly Glu Val Arg Ala Gln Leu Leu Glu Leu Pro Tyr Ala Arg Lys
210 215 220
Glu Leu Ser Leu Leu Val Leu Leu Pro Asp Asp Gly Val Glu Leu Ser
225 230 235 240
Thr Val Glu Lys Ser Leu Thr Phe Glu Lys Leu Thr Ala Trp Thr Lys
245 250 255
Pro Asp Cys Met Lys Ser Thr Glu Val Glu Val Leu Leu Pro Lys Phe
260 265 270
Lys Leu Gln Glu Asp Tyr Asp Met Glu Ser Val Leu Arg His Leu Gly
275 280 285
Ile Val Asp Ala Phe Gln Gln Gly Lys Ala Asp Leu Ser Ala Met Ser
290 295 300
Ala Glu Arg Asp Leu Cys Leu Ser Lys Phe Val His Lys Ser Phe Val
305 310 315 320
Glu Val Asn Glu Glu Gly Thr Glu Ala Ala Ala Ala Ser Ser Cys Phe
325 330 335
Val Val Ala Glu Cys Cys Met Glu Ser Gly Pro Arg Phe Cys Ala Asp
340 345 350
His Pro Phe Leu Phe Phe Ile Arg His Asn Arg Ala Asn Ser Ile Leu
355 360 365
Phe Cys Gly Arg Phe Ser Ser Pro
370 375
<210>27
<211>376
<212>PRT
<213>Homo sapiens
<220>
<223>Placental thrombin inhibitor SERPINB6
<400>27
Met Asp Val Leu Ala Glu Ala Asn Gly Thr Phe Ala Leu Asn Leu Leu
1 5 10 15
Lys Thr Leu Gly Lys Asp Asn Ser Lys Asn Val Phe Phe Ser Pro Met
20 25 30
Ser Met Ser Cys Ala Leu Ala Met Val Tyr Met Gly Ala Lys Gly Asn
35 40 45
Thr Ala Ala Gln Met Ala Gln Ile Leu Ser Phe Asn Lys Ser Gly Gly
50 55 60
Gly Gly Asp Ile His Gln Gly Phe Gln Ser Leu Leu Thr Glu Val Asn
65 70 75 80
Lys Thr Gly Thr Gln Tyr Leu Leu Arg Val Ala Asn Arg Leu Phe Gly
85 90 95
Glu Lys Ser Cys Asp Phe Leu Ser Ser Phe Arg Asp Ser Cys Gln Lys
100 105 110
Phe Tyr Gln Ala Glu Met Glu Glu Leu Asp Phe Ile Ser Ala Val Glu
115 120 125
Lys Ser Arg Lys His Ile Asn Thr Trp Val Ala Glu Lys Thr Glu Gly
130 135 140
Lys Ile Ala Glu Leu Leu Ser Pro Gly Ser Val Asp Pro Leu Thr Arg
145 150 155 160
Leu Val Leu Val Asn Ala Val Tyr Phe Arg Gly Asn Trp Asp Glu Gln
165 170 175
Phe Asp Lys Glu Asn Thr Glu Glu Arg Leu Phe Lys Val Ser Lys Asn
180 185 190
Glu Glu Lys Pro Val Gln Met Met Phe Lys Gln Ser Thr Phe Lys Lys
195 200 205
Thr Tyr Ile Gly Glu Ile Phe Thr Gln Ile Leu Val Leu Pro Tyr Val
210 215 220
Gly Lys Glu Leu Asn Met Ile Ile Met Leu Pro Asp Glu Thr Thr Asp
225 230 235 240
Leu Arg Thr Val Glu Lys Glu Leu Thr Tyr Glu Lys Phe Val Glu Trp
245 250 255
Thr Arg Leu Asp Met Met Asp Glu Glu Glu Val Glu Val Ser Leu Pro
260 265 270
Arg Phe Lys Leu Glu Glu Ser Tyr Asp Met Glu Ser Val Leu Arg Asn
275 280 285
Leu Gly Met Thr Asp Ala Phe Glu Leu Gly Lys Ala Asp Phe Ser Gly
290 295 300
Met Ser Gln Thr Asp Leu Ser Leu Ser Lys Val Val His Lys Ser Phe
305 310 315 320
Val Glu Val Asn Glu Glu Gly Thr Glu Ala Ala Ala Ala Thr Ala Ala
325 330 335
Ile Met Met Met Arg Cys Ala Arg Phe Val Pro Arg Phe Cys Ala Asp
340 345 350
His Pro Phe Leu Phe Phe Ile Gln His Ser Lys Thr Asn Gly Ile Leu
355 360 365
Phe Cys Gly Arg Phe Ser Ser Pro
370 375
<210>28
<211>485
<212>PRT
<213>Homo sapiens
<220>
<223>Angiotensinogen precursor SERPINA8
<400>28
Met Arg Lys Arg Ala Pro Gln Ser Glu Met Ala Pro Ala Gly Val Ser
1 5 10 15
Leu Arg Ala Thr Ile Leu Cys Leu Leu Ala Trp Ala Gly Leu Ala Ala
20 25 30
Gly Asp Arg Val Tyr Ile His Pro Phe His Leu Val Ile His Asn Glu
35 40 45
Ser Thr Cys Glu Gln Leu Ala Lys Ala Asn Ala Gly Lys Pro Lys Asp
50 55 60
Pro Thr Phe Ile Pro Ala Pro Ile Gln Ala Lys Thr Ser Pro Val Asp
65 70 75 80
Glu Lys Ala Leu Gln Asp Gln Leu Val Leu Val Ala Ala Lys Leu Asp
85 90 95
Thr Glu Asp Lys Leu Arg Ala Ala Met Val Gly Met Leu Ala Asn Phe
100 105 110
Leu Gly Phe Arg Ile Tyr Gly Met His Ser Glu Leu Trp Gly Val Val
115 120 125
His Gly Ala Thr Val Leu Ser Pro Thr Ala Val Phe Gly Thr Leu Ala
130 135 140
Ser Leu Tyr Leu Gly Ala Leu Asp His Thr Ala Asp Arg Leu Gln Ala
145 150 155 160
Ile Leu Gly Val Pro Trp Lys Asp Lys Asn Cys Thr Ser Arg Leu Asp
165 170 175
Ala His Lys Val Leu Ser Ala Leu Gln Ala Val Gln Gly Leu Leu Val
180 185 190
Ala Gln Gly Arg Ala Asp Ser Gln Ala Gln Leu Leu Leu Ser Thr Val
195 200 205
Val Gly Val Phe Thr Ala Pro Gly Leu His Leu Lys Gln Pro Phe Val
210 215 220
Gln Gly Leu Ala Leu Tyr Thr Pro Val Val Leu Pro Arg Ser Leu Asp
225 230 235 240
Phe Thr Glu Leu Asp Val Ala Ala Glu Lys Ile Asp Arg Phe Met Gln
245 250 255
Ala Val Thr Gly Trp Lys Thr Gly Cys Ser Leu Met Gly Ala Ser Val
260 265 270
Asp Ser Thr Leu Ala Phe Asn Thr Tyr Val His Phe Gln Gly Lys Met
275 280 285
Lys Gly Phe Ser Leu Leu Ala Glu Pro Gln Glu Phe Trp Val Asp Asn
290 295 300
Ser Thr Ser Val Ser Val Pro Met Leu Ser Gly Met Gly Thr Phe Gln
305 310 315 320
His Trp Ser Asp Ile Gln Asp Asn Phe Ser Val Thr Gln Val Pro Phe
325 330 335
Thr Glu Ser Ala Cys Leu Leu Leu Ile Gln Pro His Tyr Ala Ser Asp
340 345 350
Leu Asp Lys Val Glu Gly Leu Thr Phe Gln Gln Asn Ser Leu Asn Trp
355 360 365
Met Lys Lys Leu Ser Pro Arg Thr Ile His Leu Thr Met Pro Gln Leu
370 375 380
Val Leu Gln Gly Ser Tyr Asp Leu Gln Asp Leu Leu Ala Gln Ala Glu
385 390 395 400
Leu Pro Ala Ile Leu His Thr Glu Leu Asn Leu Gln Lys Leu Ser Asn
405 410 415
Asp Arg Ile Arg Val Gly Glu Val Leu Asn Ser Ile Phe Phe Glu Leu
420 425 430
Glu Ala Asp Glu Arg Glu Pro Thr Glu Ser Thr Gln Gln Leu Asn Lys
435 440 445
Pro Glu Val Leu Glu Val Thr Leu Asn Arg Pro Phe Leu Phe Ala Val
450 455 460
Tyr Asp Gln Ser Ala Thr Ala Leu His Phe Leu Gly Arg Val Ala Asn
465 470 475 480
Pro Leu Ser Thr Ala
485
<210>29
<211>405
<212>PRT
<213>Homo sapiens
<220>
<223>Corticosteroid-binding globulin precursor SERPINA6
<400>29
Met Pro Leu Leu Leu Tyr Thr Cys Leu Leu Trp Leu Pro Thr Ser Gly
1 5 10 15
Leu Trp Thr Val Gln Ala Met Asp Pro Asn Ala Ala Tyr Val Asn Met
20 25 30
Ser Asn His His Arg Gly Leu Ala Ser Ala Asn Val Asp Phe Ala Phe
35 40 45
Ser Leu Tyr Lys His Leu Val Ala Leu Ser Pro Lys Lys Asn Ile Phe
50 55 60
Ile Ser Pro Val Ser Ile Ser Met Ala Leu Ala Met Leu Ser Leu Gly
65 70 75 80
Thr Cys Gly His Thr Arg Ala Gln Leu Leu Gln Gly Leu Gly Phe Asn
85 90 95
Leu Thr Glu Arg Ser Glu Thr Glu Ile His Gln Gly Phe Gln His Leu
100 105 110
His Gln Leu Phe Ala Lys Ser Asp Thr Ser Leu Glu Met Thr Met Gly
115 120 125
Asn Ala Leu Phe Leu Asp Gly Ser Leu Glu Leu Leu Glu Ser Phe Ser
130 135 140
Ala Asp Ile Lys His Tyr Tyr Glu Ser Glu Val Leu Ala Met Asn Phe
145 150 155 160
Gln Asp Trp Ala Thr Ala Ser Arg Gln Ile Asn Ser Tyr Val Lys Asn
165 170 175
Lys Thr Gln Gly Lys Ile Val Asp Leu Phe Ser Gly Leu Asp Ser Pro
180 185 190
Ala Ile Leu Val Leu Val Asn Tyr Ile Phe Phe Lys Gly Thr Trp Thr
195 200 205
Gln Pro Phe Asp Leu Ala Ser Thr Arg Glu Glu Asn Phe Tyr Val Asp
210 215 220
Glu Thr Thr Val Val Lys Val Pro Met Met Leu Gln Ser Ser Thr Ile
225 230 235 240
Ser Tyr Leu His Asp Ser Glu Leu Pro Cys Gln Leu Val Gln Met Asn
245 250 255
Tyr Val Gly Asn Gly Thr Val Phe Phe Ile Leu Pro Asp Lys Gly Lys
260 265 270
Met Asn Thr Val Ile Ala Ala Leu Ser Arg Asp Thr Ile Asn Arg Trp
275 280 285
Ser Ala Gly Leu Thr Ser Ser Gln Val Asp Leu Tyr Ile Pro Lys Val
290 295 300
Thr Ile Ser Gly Val Tyr Asp Leu Gly Asp Val Leu Glu Glu Met Gly
305 310 315 320
Ile Ala Asp Leu Phe Thr Asn Gln Ala Asn Phe Ser Arg Ile Thr Gln
325 330 335
Asp Ala Gln Leu Lys Ser Ser Lys Val Val His Lys Ala Val Leu Gln
340 345 350
Leu Asn Glu Glu Gly Val Asp Thr Ala Gly Ser Thr Gly Val Thr Leu
355 360 365
Asn Leu Thr Ser Lys Pro Ile Ile Leu Arg Phe Asn Gln Pro Phe Ile
370 375 380
Ile Met Ile Phe Asp His Phe Thr Trp Ser Ser Leu Phe Leu Ala Arg
385 390 395 400
Val Met Asn Pro Val
405
<210>30
<211>417
<212>PRT
<213>Homo sapiens
<220>
<223>47 kDa heat shock protein precursor SERPINH1
<400>30
Met Arg Ser Leu Leu Leu Gly Thr Leu Cys Leu Leu Ala Val Ala Leu
1 5 10 15
Ala Ala Glu Val Lys Lys Pro Val Glu Ala Ala Ala Pro Gly Thr Ala
20 25 30
Glu Lys Leu Ser Ser Lys Ala Thr Thr Leu Ala Glu Pro Ser Thr Gly
35 40 45
Leu Ala Phe Ser Leu Tyr Gln Ala Met Ala Lys Asp Gln Ala Val Glu
50 55 60
Asn Ile Leu Val Ser Pro Val Val Val Ala Ser Ser Leu Gly Leu Val
65 70 75 80
Ser Leu Gly Gly Lys Ala Thr Thr Ala Ser Gln Ala Lys Ala Val Leu
85 90 95
Ser Ala Glu Gln Leu Arg Asp Glu Glu Val His Ala Gly Leu Gly Glu
100 105 110
Leu Leu Arg Ser Leu Ser Asn Ser Thr Ala Arg Asn Val Thr Trp Lys
115 120 125
Leu Gly Ser Arg Leu Tyr Gly Pro Ser Ser Val Ser Phe Ala Asp Asp
130 135 140
Phe Val Arg Ser Ser Lys Gln His Tyr Asn Cys Glu His Ser Lys Ile
145 150 155 160
Asn Phe Pro Asp Lys Arg Ser Ala Leu Gln Ser Ile Asn Glu Trp Ala
165 170 175
Ala Gln Thr Thr Asp Gly Lys Leu Pro Glu Val Thr Lys Asp Val Glu
180 185 190
Arg Thr Asp Gly Ala Leu Leu Val Asn Ala Met Phe Phe Lys Pro His
195 200 205
Trp Asp Glu Lys Phe His His Lys Met Val Asp Asn Arg Gly Phe Met
210 215 220
Val Thr Arg Ser Tyr Thr Val Gly Val Thr Met Met His Arg Thr Gly
225 230 235 240
Leu Tyr Asn Tyr Tyr Asp Asp Glu Lys Glu Lys Leu Gln Leu Val Glu
245 250 255
Met Pro Leu Ala His Lys Leu Ser Ser Leu Ile Ile Leu Met Pro His
260 265 270
His Val Glu Pro Leu Glu Arg Leu Glu Lys Leu Leu Thr Lys Glu Gln
275 280 285
Leu Lys Ile Trp Met Gly Lys Met Gln Lys Lys Ala Val Ala Ile Ser
290 295 300
Leu Pro Lys Gly Val Val Glu Val Thr His Asp Leu Gln Lys His Leu
305 310 315 320
Ala Gly Leu Gly Leu Thr Glu Ala Ile Asp Lys Asn Lys Ala Asp Leu
325 330 335
Ser Arg Met Ser Gly Lys Lys Asp Leu Tyr Leu Ala Ser Val Phe His
340 345 350
Ala Thr Ala Phe Glu Leu Asp Thr Asp Gly Asn Pro Phe Asp Gln Asp
355 360 365
Ile Tyr Gly Arg Glu Glu Leu Arg Ser Pro Lys Leu Phe Tyr Ala Asp
370 375 380
His Pro Phe Ile Phe Leu Val Arg Asp Thr Gln Ser Gly Ser Leu Leu
385 390 395 400
Phe Ile Gly Arg Leu Val Arg Leu Lys Gly Asp Lys Met Arg Asp Glu
405 410 415
Leu
<210>31
<211>418
<212>PRT
<213>Homo sapiens
<220>
<223>Pigment epithelium-derived factor precursor
SERPINF1
<400>31
Met Gln Ala Leu Val Leu Leu Leu Cys Ile Gly Ala Leu Leu Gly His
1 5 10 15
Ser Ser Cys Gln Asn Pro Ala Ser Pro Pro Glu Glu Gly Ser Pro Asp
20 25 30
Pro Asp Ser Thr Gly Ala Leu Val Glu Glu Glu Asp Pro Phe Phe Lys
35 40 45
Val Pro Val Asn Lys Leu Ala Ala Ala Val Ser Asn Phe Gly Tyr Asp
50 55 60
Leu Tyr Arg Val Arg Ser Ser Met Ser Pro Thr Thr Asn Val Leu Leu
65 70 75 80
Ser Pro Leu Ser Val Ala Thr Ala Leu Ser Ala Leu Ser Leu Gly Ala
85 90 95
Glu Gln Arg Thr Glu Ser Ile Ile His Arg Ala Leu Tyr Tyr Asp Leu
100 105 110
Ile Ser Ser Pro Asp Ile His Gly Thr Tyr Lys Glu Leu Leu Asp Thr
115 120 125
Val Thr Ala Pro Gln Lys Asn Leu Lys Ser Ala Ser Arg Ile Val Phe
130 135 140
Glu Lys Lys Leu Arg Ile Lys Ser Ser Phe Val Ala Pro Leu Glu Lys
145 150 155 160
Ser Tyr Gly Thr Arg Pro Arg Val Leu Thr Gly Asn Pro Arg Leu Asp
165 170 175
Leu Gln Glu Ile Asn Asn Trp Val Gln Ala Gln Met Lys Gly Lys Leu
180 185 190
Ala Arg Ser Thr Lys Glu Ile Pro Asp Glu Ile Ser Ile Leu Leu Leu
195 200 205
Gly Val Ala His Phe Lys Gly Gln Trp Val Thr Lys Phe Asp Ser Arg
210 215 220
Lys Thr Ser Leu Glu Asp Phe Tyr Leu Asp Glu Glu Arg Thr Val Arg
225 230 235 240
Val Pro Met Met Ser Asp Pro Lys Ala Val Leu Arg Tyr Gly Leu Asp
245 250 255
Ser Asp Leu Ser Cys Lys Ile Ala Gln Leu Pro Leu Thr Gly Ser Met
260 265 270
Ser Ile Ile Phe Phe Leu Pro Leu Lys Val Thr Gln Asn Leu Thr Leu
275 280 285
Ile Glu Glu Ser Leu Thr Ser Glu Phe Ile His Asp Ile Asp Arg Glu
290 295 300
Leu Lys Thr Val Gln Ala Val Leu Thr Val Pro Lys Leu Lys Leu Ser
305 310 315 320
Tyr Glu Gly Glu Val Thr Lys Ser Leu Gln Glu Met Lys Leu Gln Ser
325 330 335
Leu Phe Asp Ser Pro Asp Phe Ser Lys Ile Thr Gly Lys Pro Ile Lys
340 345 350
Leu Thr Gln Val Glu His Arg Ala Gly Phe Glu Trp Asn Glu Asp Gly
355 360 365
Ala Gly Thr Thr Pro Ser Pro Gly Leu Gln Pro Ala His Leu Thr Phe
370 375 380
Pro Leu Asp Tyr His Leu Asn Gln Pro Phe Ile Phe Val Leu Arg Asp
385 390 395 400
Thr Asp Thr Gly Ala Leu Leu Phe Ile Gly Lys Ile Leu Asp Pro Arg
405 410 415
Gly Pro
<210>32
<211>375
<212>PRT
<213>Homo sapiens
<220>
<223>Maspin SERPINB5
<400>32
Met Asp Ala Leu Gln Leu Ala Asn Ser Ala Phe Ala Val Asp Leu Phe
1 5 10 15
Lys Gln Leu Cys Glu Lys Glu Pro Leu Gly Asn Val Leu Phe Ser Pro
20 25 30
Ile Cys Leu Ser Thr Ser Leu Ser Leu Ala Gln Val Gly Ala Lys Gly
35 40 45
Asp Thr Ala Asn Glu Ile Gly Gln Val Leu His Phe Glu Asn Val Lys
50 55 60
Asp Ile Pro Phe Gly Phe Gln Thr Val Thr Ser Asp Val Asn Lys Leu
65 70 75 80
Ser Ser Phe Tyr Ser Leu Lys Leu Ile Lys Arg Leu Tyr Val Asp Lys
85 90 95
Ser Leu Asn Leu Ser Thr Glu Phe Ile Ser Ser Thr Lys Arg Pro Tyr
100 105 110
Ala Lys Glu Leu Glu Thr Val Asp Phe Lys Asp Lys Leu Glu Glu Thr
115 120 125
Lys Gly Gln Ile Asn Asn Ser Ile Lys Asp Leu Thr Asp Gly His Phe
130 135 140
Glu Asn Ile Leu Ala Asp Asn Ser Val Asn Asp Gln Thr Lys Ile Leu
145 150 155 160
Val Val Asn Ala Ala Tyr Phe Val Gly Lys Trp Met Lys Lys Phe Pro
165 170 175
Glu Ser Glu Thr Lys Glu Cys Pro Phe Arg Leu Asn Lys Thr Asp Thr
180 185 190
Lys Pro Val Gln Met Met Asn Met Glu Ala Thr Phe Cys Met Gly Asn
195 200 205
Ile Asp Ser Ile Asn Cys Lys Ile Ile Glu Leu Pro Phe Gln Asn Lys
210 215 220
His Leu Ser Met Phe Ile Leu Leu Pro Lys Asp Val Glu Asp Glu Ser
225 230 235 240
Thr Gly Leu Glu Lys Ile Glu Lys Gln Leu Asn Ser Glu Ser Leu Ser
245 250 255
Gln Trp Thr Asn Pro Ser Thr Met Ala Asn Ala Lys Val Lys Leu Ser
260 265 270
Ile Pro Lys Phe Lys Val Glu Lys Met Ile Asp Pro Lys Ala Cys Leu
275 280 285
Glu Asn Leu Gly Leu Lys His Ile Phe Ser Glu Asp Thr Ser Asp Phe
290 295 300
Ser Gly Met Ser Glu Thr Lys Gly Val Ala Leu Ser Asn Val Ile His
305 310 315 320
Lys Val Cys Leu Glu Ile Thr Glu Asp Gly Gly Asp Ser Ile Glu Val
325 330 335
Pro Gly Ala Arg Ile Leu Gln His Lys Asp Glu Leu Asn Ala Asp His
340 345 350
Pro Phe Ile Tyr Ile Ile Arg His Asn Lys Thr Arg Asn Ile Ile Phe
355 360 365
Phe Gly Lys Phe Cys Ser Pro
370 375
<210>33
<211>418
<212>PRT
<213>Homo sapiens
<220>
<223>Collagen-binding protein 2 precursor SERPINH2
<400>33
Met Arg Ser Leu Leu Leu Leu Ser Ala Phe Cys Leu Leu Glu Ala Ala
1 5 10 15
Leu Ala Ala Glu Val Lys Lys Pro Ala Ala Ala Ala Ala Pro Gly Thr
20 25 30
Ala Glu Lys Leu Ser Pro Lys Ala Ala Thr Leu Ala Glu Arg Ser Ala
35 40 45
Gly Leu Ala Phe Ser Leu Tyr Gln Ala Met Ala Lys Asp Gln Ala Val
50 55 60
Glu Asn Ile Leu Val Ser Pro Val Val Val Ala Ser Ser Leu Gly Leu
65 70 75 80
Val Ser Leu Gly Gly Lys Ala Thr Thr Ala Ser Gln Ala Lys Ala Val
85 90 95
Leu Ser Ala Glu Gln Leu Arg Asp Glu Glu Val His Ala Gly Leu Gly
100 105 110
Glu Leu Leu Arg Ser Leu Ser Asn Ser Thr Ala Arg Asn Val Thr Trp
115 120 125
Lys Leu Gly Ser Arg Leu Tyr Gly Pro Ser Ser Val Ser Phe Ala Asp
130 135 140
Asp Phe Val Arg Ser Ser Lys Gln His Tyr Asn Cys Glu His Ser Lys
145 150 155 160
Ile Asn Phe Arg Asp Lys Arg Ser Ala Leu Gln Ser Ile Asn Glu Trp
165 170 175
Ala Ala Gln Thr Thr Asp Gly Lys Leu Pro Glu Val Thr Lys Asp Val
180 185 190
Glu Arg Thr Asp Gly Ala Leu Leu Val Asn Ala Met Phe Phe Lys Pro
195 200 205
His Trp Asp Glu Lys Phe His His Lys Met Val Asp Asn Arg Gly Phe
210 215 220
Met Val Thr Arg Ser Tyr Thr Val Gly Val Met Met Met His Arg Thr
225 230 235 240
Gly Leu Tyr Asn Tyr Tyr Asp Asp Glu Lys Glu Lys Leu Gln Ile Val
245 250 255
Glu Met Pro Leu Ala His Lys Leu Ser Ser Leu Ile Ile Leu Met Pro
260 265 270
His His Val Glu Pro Leu Glu Arg Leu Glu Lys Leu Leu Thr Lys Glu
275 280 285
Gln Leu Lys Ile Trp Met Gly Lys Met Gln Lys Lys Ala Val Ala Ile
290 295 300
Ser Leu Pro Lys Gly ValVal Glu Val Thr His Asp Leu Gln Lys His
305 310 315 320
Leu Ala Gly Leu Gly Leu Thr Glu Ala Ile Asp Lys Asn Lys Ala Asp
325 330 335
Leu Ser Arg Met Ser Gly Lys Lys Asp Leu Tyr Leu Ala Ser Val Phe
340 345 350
His Ala Thr Ala Phe Glu Leu Asp Thr Asp Gly Asn Pro Phe Asp Gln
355 360 365
Asp Ile Tyr Gly Arg Glu Glu Leu Arg Ser Pro Lys Leu Phe Tyr Ala
370 375 380
Asp His Pro Phe Ile Phe Leu Val Arg Asp Thr Gln Ser Gly Ser Leu
385 390 395 400
Leu Phe Ile Gly Arg Leu Val Arg Pro Lys Gly Asp Lys Met Arg Asp
405 410 415
Glu Leu
<210>34
<211>415
<212>PRT
<213>Homo sapiens
<220>
<223>Thyroxine-binding globulin precursor SERPINA7
<400>34
Met Ser Pro Phe Leu Tyr Leu Val Leu Leu Val Leu Gly Leu His Ala
1 5 10 15
Thr Ile His Cys Ala Ser Pro Glu Gly Lys Val Thr Ala Cys His Ser
20 25 30
Ser Gln Pro Asn Ala Thr Leu Tyr Lys Met Ser Ser Ile Asn Ala Asp
35 40 45
Phe Ala Phe Asn Leu Tyr Arg Arg Phe Thr Val Glu Thr Pro Asp Lys
50 55 60
Asn Ile Phe Phe Ser Pro Val Ser Ile Ser Ala Ala Leu Val Met Leu
65 70 75 80
Ser Phe Gly Ala Cys Cys Ser Thr Gln Thr Glu Ile Val Glu Thr Leu
85 90 95
Gly Phe Asn Leu Thr Asp Thr Pro Met Val Glu Ile Gln His Gly Phe
100 105 110
Gln His Leu Ile Cys Ser Leu Asn Phe Pro Lys Lys Glu Leu Glu Leu
115 120 125
Gln Ile Gly Asn Ala Leu Phe Ile Gly Lys His Leu Lys Pro Leu Ala
130 135 140
Lys Phe Leu Asn Asp Val Lys Thr Leu Tyr Glu Thr Glu Val Phe Ser
145 150 155 160
Thr Asp Phe Ser Asn Ile Ser Ala Ala Lys Gln Glu Ile Asn Ser His
165 170 175
Val Glu Met Gln Thr Lys Gly Lys Val Val Gly Leu Ile Gln Asp Leu
180 185 190
Lys Pro Asn Thr Ile Met Val Leu Val Asn Tyr Ile His Phe Lys Ala
195 200 205
Gln Trp Ala Asn Pro Phe Asp Pro Ser Lys Thr Glu Asp Ser Ser Ser
210 215 220
Phe Leu Ile Asp Lys Thr Thr Thr Val Gln Val Pro Met Met His Gln
225 230 235 240
Met Glu Gln Tyr Tyr His Leu Val Asp Met Glu Leu Asn Cys Thr Val
245 250 255
Leu Gln Met Asp Tyr Ser Lys Asn Ala Leu Ala Leu Phe Val Leu Pro
260 265 270
Lys Glu Gly Gln Met Glu Ser Val Glu Ala Ala Met Ser Ser Lys Thr
275 280 285
Leu Lys Lys Trp Asn Arg Leu Leu Gln Lys Gly Trp Val Asp Leu Phe
290 295 300
Val Pro Lys Phe Ser Ile Ser Ala Thr Tyr Asp Leu Gly Ala Thr Leu
305 310 315 320
Leu Lys Met Gly Ile Gln His Ala Tyr Ser Glu Asn Ala Asp Phe Ser
325 330 335
Gly Leu Thr Glu Asp Asn Gly Leu Lys Leu Ser Asn Ala Ala His Lys
340 345 350
Ala Val Leu His Ile Gly Glu Lys Gly Thr Glu Ala Ala Ala Val Pro
355 360 365
Glu Val Glu Leu Ser Asp Gln Pro Glu Asn Thr Phe Leu His Pro Ile
370 375 380
Ile Gln Ile Asp Arg Ser Phe Met Leu Leu Ile Leu Glu Arg Ser Thr
385 390 395 400
Arg Ser Ile Leu Phe Leu Gly Lys Val Val Asn Pro Thr Glu Ala
405 410 415
<210>35
<211>417
<212>PRT
<213>Homo sapiens
<220>
<223>Germinal center B-cell expressed transcript 1
protein
SERPINA9
<400>35
Met Ala Ser Tyr Leu Tyr Gly Val Leu Phe Ala Val Gly Leu Cys Ala
1 5 10 15
Pro Ile Tyr Cys Val Ser Pro Ala Asn Ala Pro Ser Ala Tyr Pro Arg
20 25 30
Pro Ser Ser Thr Lys Ser Thr Pro Ala Ser Gln Val Tyr Ser Leu Asn
35 40 45
Thr Asp Phe Ala Phe Arg Leu Tyr Arg Arg Leu Val Leu Glu Thr Pro
50 55 60
Ser Gln Asn Ile Phe Phe Ser Pro Val Ser Val Ser Thr Ser Leu Ala
65 70 75 80
Met Leu Ser Leu Gly Ala His Ser Val Thr Lys Thr Gln Ile Leu Gln
85 90 95
Gly Leu Gly Phe Asn Leu Thr His Thr Pro Glu Ser Ala Ile His Gln
100 105 110
Gly Phe Gln His Leu Val His Ser Leu Thr Val Pro Ser Lys Asp Leu
115 120 125
Thr Leu Lys Met Gly Ser Ala Leu Phe Val Lys Lys Glu Leu Gln Leu
130 135 140
Gln Ala Asn Phe Leu Gly Asn Val Lys Arg Leu Tyr Glu Ala Glu Val
145 150 155 160
Phe Ser Thr Asp Phe Ser Asn Pro Ser Ile Ala Gln Ala Arg Ile Asn
165 170 175
Ser His Val Lys Lys Lys Thr Gln G1y Lys Val Val Asp Ile Ile Gln
180 185 190
Gly Leu Asp Leu Leu Thr Ala Met Val Leu Val Asn His Ile Phe Phe
195 200 205
Lys Ala Lys Trp Glu Lys Pro Phe His Pro Glu Tyr Thr Arg Lys Asn
210 215 220
Phe Pro Phe Leu Val Gly Glu Gln Val Thr Val His Val Pro Met Met
225 230 235 240
His Gln Lys Glu Gln Phe Ala Phe Gly Val Asp Thr Glu Leu Asn Cys
245 250 255
Phe Val Leu Gln Met Asp Tyr Lys Gly Asp Ala Val Ala Phe Phe Val
260 265 270
Leu Pro Ser Lys Gly Lys Met Arg Gln Leu Glu Gln Ala Leu Ser Ala
275 280 285
Arg Thr Leu Arg Lys Trp Ser His Ser Leu Gln Lys Arg Trp Ile Glu
290 295 300
Val Phe Ile Pro Arg Phe Ser Ile Ser Ala Ser Tyr Asn Leu Glu Thr
305 310 315 320
Ile Leu Pro Lys Met Gly Ile Gln Asn Ala Phe Asp Lys Asn Ala Asp
325 330 335
Phe Ser Gly Ile Ala Lys Arg Asp Ser Leu Gln Val Ser Lys Ala Thr
340 345 350
His Lys Ala Val Leu Asp Val Ser Glu Glu Gly Thr Glu Ala Thr Ala
355 360 365
Ala Thr Thr Thr Lys Phe Ile Val Arg Ser Lys Asp Gly Pro Ser Tyr
370 375 380
Phe Thr Val Ser Phe Asn Arg Thr Phe Leu Met Met Ile Thr Asn Lys
385 390 395 400
Ala Thr Asp Gly Ile Leu Phe Leu Gly Lys Val Glu Asn Pro Thr Lys
405 410 415
Ser
<210>36
<211>414
<212>PRT
<213>Homo sapiens
<220>
<223>Vaspin SERPINA12
<400>36
Met Asn Pro Thr Leu Gly Leu Ala Ile Phe Leu Ala Val Leu Leu Thr
1 5 10 15
Val Lys Gly Leu Leu Lys Pro Ser Phe Ser Pro Arg Asn Tyr Lys Ala
20 25 30
Leu Ser Glu Val Gln Gly Trp Lys Gln Arg Met Ala Ala Lys Glu Leu
35 40 45
Ala Arg Gln Asn Met Asp Leu Gly Phe Lys Leu Leu Lys Lys Leu Ala
50 55 60
Phe Tyr Asn Pro Gly Arg Asn Ile Phe Leu Ser Pro Leu Ser Ile Ser
65 70 75 80
Thr Ala Phe Ser Met Leu Cys Leu Gly Ala Gln Asp Ser Thr Leu Asp
85 90 95
Glu Ile Lys Gln G1y Phe Asn Phe Arg Lys Met Pro Glu Lys Asp Leu
100 105 110
His Glu Gly Phe His Tyr Ile Ile His Glu Leu Thr Gln Lys Thr Gln
115 120 125
Asp Leu Lys Leu Ser Ile Gly Asn Thr Leu Phe Ile Asp Gln Arg Leu
130 135 140
Gln Pro Gln Arg Lys Phe Leu Glu Asp Ala Lys Asn Phe Tyr Ser Ala
145 150 155 160
Glu Thr Ile Leu Thr Asn Phe Gln Asn Leu Glu Met Ala Gln Lys Gln
165 170 175
Ile Asn Asp Phe Ile Ser Gln Lys Thr His Gly Lys Ile Asn Asn Leu
180 185 190
Ile Glu Asn Ile Asp Pro Gly Thr Val Met Leu Leu Ala Asn Tyr Ile
195 200 205
Phe Phe Arg Ala Arg Trp Lys His Glu Phe Asp Pro Asn Val Thr Lys
210 215 220
Glu Glu Asp Phe Phe Leu Glu Lys Asn Ser Ser Val Lys Val Pro Met
225 230 235 240
Met Phe Arg Ser Gly Ile Tyr Gln Val Gly Tyr Asp Asp Lys Leu Ser
245 250 255
Cys Thr Ile Leu Glu Ile Pro Tyr Gln Lys Asn Ile Thr Ala Ile Phe
260 265 270
Ile Leu Pro Asp Glu Gly Lys Leu Lys His Leu Glu Lys Gly Leu Gln
275 280 285
Val Asp Thr Phe Ser Arg Trp Lys Thr Leu Leu Ser Arg Arg Val Val
290 295 300
Asp Val Ser Val Pro Arg Leu His Met Thr Gly Thr Phe Asp Leu Lys
305 310 315 320
Lys Thr Leu Ser Tyr Ile Gly Val Ser Lys Ile Phe Glu Glu His Gly
325 330 335
Asp Leu Thr Lys Ile Ala Pro His Arg Ser Leu Lys Val Gly Glu Ala
340 345 350
Val His Lys Ala Glu Leu Lys Met Asp Glu Arg Gly Thr Glu Gly Ala
355 360 365
Ala Gly Thr Gly Ala Gln Thr Leu Pro Met Glu Thr Pro Leu Val Val
370 375 380
Lys Ile Asp Lys Pro Tyr Leu Leu Leu Ile Tyr Ser Glu Lys Ile Pro
385 390 395 400
Ser Val Leu Phe Leu Gly Lys Ile Val Asn Pro Ile Gly Lys
405 410
<210>37
<211>422
<212>PRT
<213>Homo sapiens
<220>
<223>Serpin A11 precursor SERPINA11
<400>37
Met Gly Pro Ala Trp Leu Trp Leu Leu Gly Thr Gly Ile Leu Ala Ser
1 5 10 15
Val His Cys Gln Pro Leu Leu Ala His Gly Asp Lys Ser Leu Gln Gly
20 25 30
Pro Gln Pro Pro Arg His Gln Leu Ser Glu Pro Ala Pro Ala Tyr His
35 40 45
Arg Ile Thr Pro Thr Ile Thr Asn Phe Ala Leu Arg Leu Tyr Lys Glu
50 55 60
Leu Ala Ala Asp Ala Pro Gly Asn Ile Phe Phe Ser Pro Val Ser Ile
65 70 75 80
Ser Thr Thr Leu Ala Leu Leu Ser Leu Gly Ala Gln Ala Asn Thr Ser
85 90 95
Ala Leu Ile Leu Glu Gly Leu Gly Phe Asn Leu Thr Glu Thr Pro Glu
100 105 110
Ala Asp Ile His Gln Gly Phe Arg Ser Leu Leu His Thr Leu Ala Leu
115 120 125
Pro Ser Pro Lys Leu Glu Leu Lys Val Gly Asn Ser Leu Phe Leu Asp
130 135 140
Lys Arg Leu Lys Pro Arg Gln His Tyr Leu Asp Ser Ile Lys Glu Leu
145 150 155 160
Tyr Gly Ala Phe Ala Phe Ser Ala Asn Phe Thr Asp Ser Val Thr Thr
165 170 175
Gly Arg Gln Ile Asn Asp Tyr Leu Arg Arg Gln Thr Tyr Gly Gln Val
180 185 190
Val Asp Cys Leu Pro Glu Phe Ser Gln Asp Thr Phe Met Val Leu Ala
195 200 205
Asn Tyr Ile Phe Phe Lys Ala Lys Trp Lys His Pro Phe Ser Arg Tyr
210 215 220
Gln Thr Gln Lys Gln Glu Ser Phe Phe Val Asp Glu Arg Thr Ser Leu
225 230 235 240
Gln Val Pro Met Met His Gln Lys Glu Met His Arg Phe Leu Tyr Asp
245 250 255
Gln Asp Leu Ala Cys Thr Val Leu Gln Ile Glu Tyr Arg Gly Asn Ala
260 265 270
Leu Ala Leu Leu Val Leu Pro Asp Pro Gly Lys Met Lys Gln Val Glu
275 280 285
Ala Ala Leu Gln Pro Gln Thr Leu Arg Lys Trp Gly Gln Leu Leu Leu
290 295 300
Pro Ser Leu Leu Asp Leu His Leu Pro Arg Phe Ser Ile Ser Gly Thr
305 310 315 320
Tyr Asn Leu Glu Asp Ile Leu Pro Gln Ile Gly Leu Thr Asn Ile Leu
325 330 335
Asn Leu Glu Ala Asp Phe Ser Gly Val Thr Gly Gln Leu Asn Lys Thr
340 345 350
Ile Ser Lys Val Ser His Lys Ala Met Val Asp Met Ser Glu Lys Gly
355 360 365
Thr Glu Ala Gly Ala Ala Ser Gly Leu Leu Ser Gln Pro Pro Ser Leu
370 375 380
Asn Thr Met Ser Asp Pro His Ala His Phe Asn Arg Pro Phe Leu Leu
385 390 395 400
Leu Leu Trp Glu Val Thr Thr Gln Ser Leu Leu Phe Leu Gly Lys Val
405 410 415
Val Asn Pro Val Ala Gly
420
<210>38
<211>307
<212>PRT
<213>Homo sapiens
<220>
<223>Serpin A13 precursor SERPINA13
<400>38
Met Glu Ala Ser Arg Trp Trp Leu Leu Val Thr Val Leu Met Ala Gly
1 5 10 15
Ala His Cys Val Ala Leu Val Asp Gln Glu Ala Ser Asp Leu Ile His
20 25 30
Ser Gly Pro Gln Asp Ser Ser Pro Gly Pro Ala Leu Pro Cys His Lys
35 40 45
Ile Ser Val Ser Asn Ile Asp Phe Ala Phe Lys Leu Tyr Arg Gln Leu
50 55 60
Ala Leu Asn Ala Pro Gly Glu Asn Ile Leu Phe Phe Pro Val Ser Ile
65 70 75 80
Ser Leu Ala Leu Ala Met Leu Ser Trp Gly Ala Pro Val Ala Ser Arg
85 90 95
Thr Gln Leu Leu Glu Gly Leu Gly Phe Thr Leu Thr Val Val Pro Glu
100 105 110
Glu Glu Ile Gln Glu Gly Phe Trp Asp Leu Leu Ile Arg Leu Arg Gly
115 120 125
Gln Gly Pro Arg Leu Leu Leu Thr Met Asp Gln Arg Arg Phe Ser Gly
130 135 140
Leu Gly Ala Arg Ala Asn Gln Ser Leu Glu Glu Ala Gln Lys His Ile
145 150 155 160
Asp Glu Tyr Thr Glu Gln Gln Thr Gln Gly Lys Leu Gly Ala Trp Glu
165 170 175
Lys Asp Leu Gly Ser Glu Thr Thr Ala Val Leu Val Asn His Met Leu
180 185 190
Leu Arg Ala Glu Trp Met Lys Pro Phe Asp Ser His Ala Thr Ser Pro
195 200 205
Lys Glu Phe Phe Val Asp Glu His Ser Ala Val Trp Val Pro Met Met
210 215 220
Lys Glu Lys Ala Ser His Arg Phe Leu His Asp Arg Glu Leu Gln Cys
225 230 235 240
Ser Val Leu Arg Met Asp His Ala Gly Asn Thr Thr Thr Phe Phe Ile
245 250 255
Phe Pro Asn Arg Gly Lys Met Arg His Leu Glu Asp Ala Leu Leu Pro
260 265 270
Glu Thr Leu Ile Lys Trp Asp Ser Leu Leu Arg Thr Arg Glu Leu Asp
275 280 285
Phe His Phe Pro Lys Phe Ser Ile Ser Arg Thr Cys Arg Leu Glu Met
290 295 300
Leu Leu Pro
305
<210>39
<211>934
<212>DNA
<213>Homo sapiens
<220>
<223>granzyme B
<400>39
tgagaagatg caaccaatcc tgcttctgct ggccttcctc ctgctgccca gggcagatgc 60
aggggagatc atcgggggac atgaggccaa gccccactcc cgcccctaca tggcttatct 120
tatgatctgg gatcagaagt ctctgaagag gtgcggtggc ttcctgatac aagacgactt 180
cgtgctgaca gctgctcact gttggggaag ctccataaat gtcaccttgg gggcccacaa 240
tatcaaagaa caggagccga cccagcagtt tatccctgtg aaaagaccca tcccccatcc 300
agcctataat cctaagaact tctccaacga catcatgcta ctgcagctgg agagaaaggc 360
caagcggacc agagctgtgc agcccctcag gctacctagc aacaaggccc aggtgaagcc 420
agggcagaca tgcagtgtgg ccggctgggg gcagacggcc cccctgggaa aacactcaca 480
cacactacaa gaggtgaaga tgacagtgca ggaagatcga aagtgcgaat ctgacttacg 540
ccattattac gacagtacca ttgagttgtg cgtgggggac ccagagatta aaaagacttc 600
ctttaagggg gactctggag gccctcttgt gtgtaacaag gtggcccagg gcattgtctc 660
ctatggacga aacaatggca tgcctccacg agcctgcacc aaagtctcaa gctttgtaca 720
ctggataaag aaaaccatga aacgctacta actacaggaa gcaaactaag cccccgctgt 780
aatgaaacac cttctctgga gccaagtcca gatttacact gggagaggtg ccagcaactg 840
aataaatacc tctcccagtg taaatctgga gccaagtcca gatttacact gggagaggtg 900
ccagcaactg aataaatacc tcttagctga gtgg 934
<210>40
<211>247
<212>PRT
<213>Homo sapiens
<220>
<223>granzyme B
<400>40
Met Gln Pro Ile Leu Leu Leu Leu Ala Phe Leu Leu Leu Pro Arg Ala
1 5 10 15
Asp Ala Gly Glu Ile Ile Gly Gly His Glu Ala Lys Pro His Ser Arg
20 25 30
Pro Tyr Met Ala Tyr Leu Met Ile Trp Asp Gln Lys Ser Leu Lys Arg
35 40 45
Cys Gly Gly Phe Leu Ile Gln Asp Asp Phe Val Leu Thr Ala Ala His
50 55 60
Cys Trp Gly Ser Ser Ile Asn Val Thr Leu Gly Ala His Asn Ile Lys
65 70 75 80
Glu Gln Glu Pro Thr Gln Gln Phe Ile Pro Val Lys Arg Pro Ile Pro
85 90 95
His Pro Ala Tyr Asn Pro Lys Asn Phe Ser Asn Asp Ile Met Leu Leu
100 105 110
Gln Leu Glu Arg Lys Ala Lys Arg Thr Arg Ala Val Gln Pro Leu Arg
115 120 125
Leu Pro Ser Asn Lys Ala Gln Val Lys Pro Gly Gln Thr Cys Ser Val
130 135 140
Ala Gly Trp Gly Gln Thr Ala Pro Leu Gly Lys His Ser His Thr Leu
145 150 155 160
Gln Glu Val Lys Met Thr Val Gln Glu Asp Arg Lys Cys Glu Ser Asp
165 170 175
Leu Arg His Tyr Tyr Asp Ser Thr Ile Glu Leu Cys Val Gly Asp Pro
180 185 190
Glu Ile Lys Lys Thr Ser Phe Lys Gly Asp Ser Gly Gly Pro Leu Val
195 200 205
Cys Asn Lys Val Ala Gln Gly Ile Val Ser Tyr Gly Arg Asn Asn Gly
210 215 220
Met Pro Pro Arg Ala Cys Thr Lys Val Ser Ser Phe Val His Trp Ile
225 230 235 240
Lys Lys Thr Met Lys Arg Tyr
245
<210>41
<211>1165
<212>DNA
<213>Homo sapiens
<220>
<223>Testisin transcript variant 1
<400>41
gctgggagta gagggcagag ctcccacccc gccccgcccc cagggggcgc cccgggcccg 60
gcgcgagagg aggcagaggg ggcgtcaggc cgcgggagag gaggccatgg gcgcgcgcgg 120
ggcgctgctg ctggcgctgc tgctggctcg ggctggactc aggaagccgg agtcgcagga 180
ggcggcgccg ttatcaggac catgcggccg acgggtcatc acgtcgcgca tcgtgggtgg 240
agaggacgcc gaactcgggc gttggccgtg gcaggggagc ctgcgcctgt gggattccca 300
cgtatgcgga gtgagcctgc tcagccaccg ctgggcactc acggcggcgc actgctttga 360
aacctatagt gaccttagtg atccctccgg gtggatggtc cagtttggcc agctgacttc 420
catgccatcc ttctggagcc tgcaggccta ctacacccgt tacttcgtat cgaatatcta 480
tctgagccct cgctacctgg ggaattcacc ctatgacatt gccttggtga agctgtctgc 540
acctgtcacc tacactaaac acatccagcc catctgtctc caggcctcca catttgagtt 600
tgagaaccgg acagactgct gggtgactgg ctgggggtac atcaaagagg atgaggcact 660
gccatctccc cacaccctcc aggaagttca ggtcgccatc ataaacaact ctatgtgcaa 720
ccacctcttc ctcaagtaca gtttccgcaa ggacatcttt ggagacatgg tttgtgctgg 780
caatgcccaa ggcgggaagg atgcctgctt cggtgactca ggtggaccct tggcctgtaa 840
caagaatgga ctgtggtatc agattggagt cgtgagctgg ggagtgggct gtggtcggcc 900
caatcggccc ggtgtctaca ccaatatcag ccaccacttt gagtggatcc agaagctgat 960
ggcccagagt ggcatgtccc agccagaccc ctcctggccg ctactctttt tccctcttct 1020
ctgggctctc ccactcctgg ggccggtctg agcctacctg agcccatgca gcctggggcc 1080
actgccaagt caggccctgg ttctcttctg tcttgtttgg taataaacac attccagttg 1140
atgccttgca gggcattctt caaaa 1165
<210>42
<211>314
<212>PRT
<213>Homo sapiens
<220>
<223>Testisin isoform 1
<400>42
Met Gly Ala Arg Gly Ala Leu Leu Leu Ala Leu Leu Leu Ala Arg Ala
1 5 10 15
Gly Leu Arg Lys Pro Glu Ser Gln Glu Ala Ala Pro Leu Ser Gly Pro
20 25 30
Cys Gly Arg Arg Val Ile Thr Ser Arg Ile ValGly Gly Glu Asp Ala
35 40 45
Glu Leu Gly Arg Trp Pro Trp Gln Gly Ser Leu Arg Leu Trp Asp Ser
50 55 60
His Val Cys Gly Val Ser Leu Leu Ser His Arg Trp Ala Leu Thr Ala
65 70 75 80
Ala His Cys Phe Glu Thr Tyr Ser Asp Leu Ser Asp Pro Ser Gly Trp
85 90 95
Met Val Gln Phe Gly Gln Leu Thr Ser Met Pro Ser Phe Trp Ser Leu
100 105 110
Gln Ala Tyr Tyr Thr Arg Tyr Phe Val Ser Asn Ile Tyr Leu Ser Pro
115 120 125
Arg Tyr Leu Gly Asn Ser Pro Tyr Asp Ile Ala Leu Val Lys Leu Ser
130 135 140
Ala Pro Val Thr Tyr Thr Lys His Ile Gln Pro Ile Cys Leu Gln Ala
145 150 155 160
Ser Thr Phe Glu Phe Glu Asn Arg Thr Asp Cys Trp Val Thr Gly Trp
165 170 175
Gly Tyr Ile Lys Glu Asp Glu Ala Leu Pro Ser Pro His Thr Leu Gln
180 185 190
Glu Val Gln ValAla Ile Ile Asn Asn Ser Met Cys Asn His Leu Phe
195 200 205
Leu Lys Tyr Ser Phe Arg Lys Asp Ile Phe Gly Asp Met Val Cys Ala
210 215 220
Gly Asn Ala Gln Gly Gly Lys Asp Ala Cys Phe Gly Asp Ser Gly Gly
225 230 235 240
Pro Leu Ala Cys Asn Lys Asn Gly Leu Trp Tyr Gln Ile Gly Val Val
245 250 255
Ser Trp Gly Val Gly Cys Gly Arg Pro Asn Arg Pro Gly Val Tyr Thr
260 265 270
Asn Ile Ser His His Phe Glu Trp Ile Gln Lys Leu Met Ala Gln Ser
275 280 285
Gly Met Ser Gln Pro Asp Pro Ser Trp Pro Leu Leu Phe Phe Pro Leu
290 295 300
Leu Trp Ala Leu Pro Leu Leu Gly Pro Val
305 310
<210>43
<211>1159
<212>DNA
<213>Homo sapiens
<220>
<223>Testisin transcript variant 2
<400>43
gctgggagta gagggcagag ctcccacccc gccccgcccc cagggggcgc cccgggcccg 60
gcgcgagagg aggcagaggg ggcgtcaggc cgcgggagag gaggccatgg gcgcgcgcgg 120
ggcgctgctg ctggcgctgc tgctggctcg ggctggactc aggaagccgg agtcgcagga 180
ggcggcgccg ttatcaggac catgcggccg acgggtcatc acgtcgcgca tcgtgggtgg 240
agaggacgcc gaactcgggc gttggccgtg gcaggggagc ctgcgcctgt gggattccca 300
cgtatgcgga gtgagcctgc tcagccaccg ctgggcactc acggcggcgc actgctttga 360
aactgacctt agtgatccct ccgggtggat ggtccagttt ggccagctga cttccatgcc 420
atccttctgg agcctgcagg cctactacac ccgttacttc gtatcgaata tctatctgag 480
ccctcgctac ctggggaatt caccctatga cattgccttg gtgaagctgt ctgcacctgt 540
cacctacact aaacacatcc agcccatctg tctccaggcc tccacatttg agtttgagaa 600
ccggacagac tgctgggtga ctggctgggg gtacatcaaa gaggatgagg cactgccatc 660
tccccacacc ctccaggaag ttcaggtcgc catcataaac aactctatgt gcaaccacct 720
cttcctcaag tacagtttcc gcaaggacat ctttggagac atggtttgtg ctggcaatgc 780
ccaaggcggg aaggatgcct gcttcggtga ctcaggtgga cccttggcct gtaacaagaa 840
tggactgtgg tatcagattg gagtcgtgag ctggggagtg ggctgtggtc ggcccaatcg 900
gcccggtgtc tacaccaata tcagccacca ctttgagtgg atccagaagc tgatggccca 960
gagtggcatg tcccagccag acccctcctg gccgctactc tttttccctc ttctctgggc 1020
tctcccactc ctggggccgg tctgagccta cctgagccca tgcagcctgg ggccactgcc 1080
aagtcaggcc ctggttctct tctgtcttgt ttggtaataa acacattcca gttgatgcct 1140
tgcagggcat tcttcaaaa 1159
<210>44
<211>312
<212>PRT
<213>Homo sapiens
<220>
<223>Testisin isoform 2
<400>44
Met Gly Ala Arg Gly Ala Leu Leu Leu Ala Leu Leu Leu Ala Arg Ala
1 5 10 15
Gly Leu Arg Lys Pro Glu Ser Gln Glu Ala Ala Pro Leu Ser Gly Pro
20 25 30
Cys Gly Arg Arg Val Ile Thr Ser Arg Ile Val Gly Gly Glu Asp Ala
35 40 45
Glu Leu Gly Arg Trp Pro Trp Gln Gly Ser Leu Arg Leu Trp Asp Ser
50 55 60
His Val Cys Gly Val Ser Leu Leu Ser His Arg Trp Ala Leu Thr Ala
65 70 75 80
Ala His Cys Phe Glu Thr Asp Leu Ser Asp Pro Ser Gly Trp Met Val
85 90 95
Gln Phe Gly Gln Leu Thr Ser Met Pro Ser Phe Trp Ser Leu Gln Ala
100 105 110
Tyr Tyr Thr Arg Tyr Phe Val Ser Asn Ile Tyr Leu Ser Pro Arg Tyr
115 120 125
Leu Gly Asn Ser Pro Tyr Asp Ile Ala Leu Val Lys Leu Ser Ala Pro
130 135 140
Val Thr Tyr Thr Lys His Ile Gln Pro Ile Cys Leu Gln Ala Ser Thr
145 150 155 160
Phe Glu Phe Glu Asn Arg Thr Asp Cys Trp Val Thr Gly Trp Gly Tyr
165 170 175
Ile Lys Glu Asp Glu Ala Leu Pro Ser Pro His Thr Leu Gln Glu Val
180 185 190
Gln Val Ala Ile Ile Asn Asn Ser Met Cys Asn His Leu Phe Leu Lys
195 200 205
Tyr Ser Phe Arg Lys Asp Ile Phe Gly Asp Met Val Cys Ala Gly Asn
210 215 220
Ala Gln Gly Gly Lys Asp Ala Cys Phe Gly Asp Ser Gly Gly Pro Leu
225 230 235 240
Ala Cys Asn Lys Asn Gly Leu Trp Tyr Gln Ile Gly Val Val Ser Trp
245 250 255
Gly Val Gly Cys Gly Arg Pro Asn Arg Pro Gly Val Tyr Thr Asn Ile
260 265 270
Ser His His Phe Glu Trp Ile Gln Lys Leu Met Ala Gln Ser Gly Met
275 280 285
Ser Gln Pro Asp Pro Ser Trp Pro Leu Leu Phe Phe Pro Leu Leu Trp
290 295 300
Ala Leu Pro Leu Leu Gly Pro Val
305 310
<210>45
<211>1135
<212>DNA
<213>Homo sapiens
<220>
<223>Testisin transcript variant 3
<400>45
gctgggagta gagggcagag ctcccacccc gccccgcccc cagggggcgc cccgggcccg 60
gcgcgagagg aggcagaggg ggcgtcaggc cgcgggagag gaggccatgg gcgcgcgcgg 120
ggcgctgctg ctggcgctgc tgctggctcg ggctggactc aggaagccgg agtcgcagga 180
ggcggcgccg ttatcaggac catgcggccg acgggtcatc acgtcgcgca tcgtgggtgg 240
agaggacgcc gaactcgggc gttggccgtg gcaggggagc ctgcgcctgt gggattccca 300
cgtatgcgga gtgagcctgc tcagccaccg ctgggcactc acggcggcgc actgctttga 360
aacctatagt gaccttagtg atccctccgg gtggatggtc cagtttggcc agctgacttc 420
catgccatcc ttctggagcc tgcaggccta ctacacccgt tacttcgtat cgaatatcta 480
tctgagccct cgctacctgg ggaattcacc ctatgacatt gccttggtga agctgtctgc 540
acctgtcacc tacactaaac acatccagcc catctgtctc caggcctcca catttgagtt 600
tgagaaccgg acagactgct gggtgactgg ctgggggtac atcaaagagg atgaggcact 660
gccatctccc cacaccctcc aggaagttca ggtcgccatc ataaacaact ctatgtgcaa 720
ccacctcttc ctcaagtaca gtttccgcaa ggacatcttt ggagacatgg gtgactcagg 780
tggacccttg gcctgtaaca agaatggact gtggtatcag attggagtcg tgagctgggg 840
agtgggctgt ggtcggccca atcggcccgg tgtctacacc aatatcagcc accactttga 900
gtggatccag aagctgatgg cccagagtgg catgtcccag ccagacccct cctggccgct 960
actctttttc cctcttctct gggctctccc actcctgggg ccggtctgag cctacctgag 1020
cccatgcagc ctggggccac tgccaagtca ggccctggtt ctcttctgtc ttgtttggta 1080
ataaacacat tccagttgat gccttgcagg gcattcttca aaaaaaaaaa aaaaa 1135
<210>46
<211>300
<212>PRT
<213>Homo sapiens
<220>
<223>Testisin isoform 3
<400>46
Met Gly Ala Arg Gly Ala Leu Leu Leu Ala Leu Leu Leu Ala Arg Ala
1 5 10 15
Gly Leu Arg Lys Pro Glu Ser Gln Glu Ala Ala Pro Leu Ser Gly Pro
20 25 30
Cys Gly Arg Arg Val Ile Thr Ser Arg Ile Val Gly Gly Glu Asp Ala
35 40 45
Glu Leu Gly Arg Trp Pro Trp Gln Gly Ser Leu Arg Leu Trp Asp Ser
50 55 60
His Val Cys Gly Val Ser Leu Leu Ser His Arg Trp Ala Leu Thr Ala
65 70 75 80
Ala His Cys Phe Glu Thr Tyr Ser Asp Leu Ser Asp Pro Ser Gly Trp
85 90 95
Met ValGln Phe Gly Gln Leu Thr Ser Met Pro Ser Phe Trp Ser Leu
100 105 110
Gln Ala Tyr Tyr Thr Arg Tyr Phe Val Ser Asn Ile Tyr Leu Ser Pro
115 120 125
Arg Tyr Leu Gly Asn Ser Pro Tyr Asp Ile Ala Leu Val Lys Leu Ser
130 135 140
Ala Pro Val Thr Tyr Thr Lys His Ile Gln Pro Ile Cys Leu Gln Ala
145 150 155 160
Ser Thr Phe Glu Phe Glu Asn Arg Thr Asp Cys Trp Val Thr Gly Trp
165 170 175
Gly Tyr Ile Lys Glu Asp Glu Ala Leu Pro Ser Pro His Thr Leu Gln
180 185 190
Glu Val Gln ValAla Ile Ile Asn Asn Ser Met Cys Asn His Leu Phe
195 200 205
Leu Lys Tyr Ser Phe Arg Lys Asp Ile Phe Gly Asp Met Gly Asp Ser
210 215 220
Gly Gly Pro Leu Ala Cys Asn Lys Asn Gly Leu Trp Tyr Gln Ile Gly
225 230 235 240
Val Val Ser Trp Gly Val Gly Cys Gly Arg Pro Asn Arg Pro Gly Val
245 250 255
Tyr Thr Asn Ile Ser His His Phe Glu Trp Ile Gln Lys Leu Met Ala
260 265 270
Gln Ser Gly Met Ser Gln Pro Asp Pro Ser Trp Pro Leu Leu Phe Phe
275 280 285
Pro Leu Leu Trp Ala Leu Pro Leu Leu Gly Pro Val
290 295 300
<210>47
<211>1194
<212>DNA
<213>Homo sapiens
<220>
<223>trypstase beta 1
<400>47
ataaatgggg aggggagagc ccactgggta gaaggaacag ggagcggcca ggatgctgaa 60
tctgctgctg ctggcgctgc ccgtcctggc gagccgcgcc tacgcggccc ctgccccagg 120
ccaggccctg cagcgagtgg gcatcgtcgg gggtcaggag gcccccagga gcaagtggcc 180
ctggcaggtg agcctgagag tccacggccc atactggatg cacttctgcg ggggctccct 240
catccacccc cagtgggtgc tgaccgcagc gcactgcgtg ggaccggacg tcaaggatct 300
ggccgccctc agggtgcaac tgcgggagca gcacctctac taccaggacc agctgctgcc 360
ggtcagcagg atcatcgtgc acccacagtt ctacaccgcc cagatcggag cggacatcgc 420
cctgctggag ctggaggagc cggtgaacgt ctccagccac gtccacacgg tcaccctgcc 480
ccctgcctca gagaccttcc ccccggggat gccgtgctgg gtcactggct ggggcgatgt 540
ggacaatgat gagcgcctcc caccgccatt tcctctgaag caggtgaagg tccccataat 600
ggaaaaccac atttgtgacg caaaatacca ccttggcgcc tacacgggag acgacgtccg 660
catcgtccgt gacgacatgc tgtgtgccgg gaacacccgg agggactcat gccagggcga 720
ctccggaggg cccctggtgt gcaaggtgaa tggcacctgg ctgcaggcgg gcgtggtcag 780
ctggggcgag ggctgtgccc agcccaaccg gcctggcatc tacacccgtg tcacctacta 840
cttggactgg atccaccact atgtccccaa aaagccgtga gtcaggcctg ggttggccac 900
ctgggtcact ggaggaccaa cccctgctgt ccaaaacacc actgcttcct acccaggtgg 960
cgactgcccc ccacaccttc cctgccccgt cctgagtgcc ccttcctgtc ctaagccccc 1020
tgctctcttc tgagcccctt cccctgtcct gaggaccctt ccctatcctg agcccccttc 1080
cctgtcctaa gcctgacgcc tgcaccgggc cctccagccc tcccctgccc agatagctgg 1140
tggtgggcgc taatcctcct gagtgctgga cctcattaaa gtgcatggaa atca 1194
<210>48
<211>275
<212>PRT
<213>Homo sapiens
<220>
<223>trypstase beta 1
<400>48
Met Leu Asn Leu Leu Leu Leu Ala Leu Pro Val Leu Ala Ser Arg Ala
1 5 10 15
Tyr Ala Ala Pro Ala Pro Gly Gln Ala Leu Gln Arg Val Gly Ile Val
20 25 30
Gly Gly Gln Glu Ala Pro Arg Ser Lys Trp Pro Trp Gln Val Ser Leu
35 40 45
Arg Val His Gly Pro Tyr Trp Met His Phe Cys Gly Gly Ser Leu Ile
50 55 60
His Pro Gln Trp Val Leu Thr Ala Ala His Cys Val Gly Pro Asp Val
65 70 75 80
Lys Asp Leu Ala Ala Leu Arg Val Gln Leu Arg Glu Gln His Leu Tyr
85 90 95
Tyr Gln Asp Gln Leu Leu Pro Val Ser Arg Ile Ile Val His Pro Gln
100 105 110
Phe Tyr Thr Ala Gln Ile Gly Ala Asp Ile Ala Leu Leu Glu Leu Glu
115 120 125
Glu Pro Val Asn Val Ser Ser His Val His Thr Val Thr Leu Pro Pro
130 135 140
Ala Ser Glu Thr Phe Pro Pro Gly Met Pro Cys Trp Val Thr Gly Trp
145 150 155 160
Gly Asp Val Asp Asn Asp Glu Arg Leu Pro Pro Pro Phe Pro Leu Lys
165 170 175
Gln ValLys Val Pro Ile Met Glu Asn His Ile Cys Asp Ala Lys Tyr
180 185 190
His Leu Gly Ala Tyr Thr Gly Asp Asp Val Arg Ile Val Arg Asp Asp
195 200 205
Met Leu Cys Ala Gly Asn Thr Arg Arg Asp Ser Cys Gln Gly Asp Ser
210 215 220
Gly Gly Pro Leu Val Cys Lys Val Asn Gly Thr Trp Leu Gln Ala Gly
225 230 235 240
Val Val Ser Trp Gly Glu Gly Cys Ala Gln Pro Asn Arg Pro Gly Ile
245 250 255
Tyr Thr Arg Val Thr Tyr Tyr Leu Asp Trp Ile His His Tyr Val Pro
260 265 270
Lys Lys Pro
275
<210>49
<211>1528
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hk5
<400>49
ggaaggcaca ggcctgagaa gtctgcggct gagctgggag caaatccccc accccctacc 60
tgggggacag ggcaagtgag acctggtgag ggtggctcag caggaaggga aggagaggtg 120
tctgtgcgtc ctgcacccac atctttctct gtcccctcct tgccctgtct ggaggctgct 180
agactcctat cttctgaatt ctatagtgcc tgggtctcag cgcagtgccg atggtggccc 240
gtccttgtgg ttcctctcta cctggggaaa taaggtgcag cggccatggc tacagcaaga 300
cccccctgga tgtgggtgct ctgtgctctg atcacagcct tgcttctggg ggtcacagag 360
catgttctcg ccaacaatga tgtttcctgt gaccacccct ctaacaccgt gccctctggg 420
agcaaccagg acctgggagc tggggccggg gaagacgccc ggtcggatga cagcagcagc 480
cgcatcatca atggatccga ctgcgatatg cacacccagc cgtggcaggc cgcgctgttg 540
ctaaggccca accagctcta ctgcggggcg gtgttggtgc atccacagtg gctgctcacg 600
gccgcccact gcaggaagaa agttttcaga gtccgtctcg gccactactc cctgtcacca 660
gtttatgaat ctgggcagca gatgttccag ggggtcaaat ccatccccca ccctggctac 720
tcccaccctg gccactctaa cgacctcatg ctcatcaaac tgaacagaag aattcgtccc 780
actaaagatg tcagacccat caacgtctcc tctcattgtc cctctgctgg gacaaagtgc 840
ttggtgtctg gctgggggac aaccaagagc ccccaagtgc acttccctaa ggtcctccag 900
tgcttgaata tcagcgtgct aagtcagaaa aggtgcgagg atgcttaccc gagacagata 960
gatgacacca tgttctgcgc cggtgacaaa gcaggtagag actcctgcca gggtgattct 1020
ggggggcctg tggtctgcaa tggctccctg cagggactcg tgtcctgggg agattaccct 1080
tgtgcccggc ccaacagacc gggtgtctac acgaacctct gcaagttcac caagtggatc 1140
caggaaacca tccaggccaa ctcctgagtc atcccaggac tcagcacacc ggcatcccca 1200
cctgctgcag ggacagccct gacactcctt tcagaccctc attccttccc agagatgttg 1260
agaatgttca tctctccagc ccctgacccc atgtctcctg gactcagggt ctgcttcccc 1320
cacattgggc tgaccgtgtc tctctagttg aaccctggga acaatttcca aaactgtcca 1380
gggcgggggt tgcgtctcaa tctccctggg gcactttcat cctcaagctc agggcccatc 1440
ccttctctgc agctctgacc caaatttagt cccagaaata aactgagaag tggaaaaaaa 1500
aaaaaaaaaa aaaaaaaaaa aaaaaaaa 1528
<210>50
<211>293
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hk5
<400>50
Met Ala Thr Ala Arg Pro Pro Trp Met Trp Val Leu Cys Ala Leu Ile
1 5 10 15
Thr Ala Leu Leu Leu Gly Val Thr Glu His Val Leu Ala Asn Asn Asp
20 25 30
Val Ser Cys Asp His Pro Ser Asn Thr Val Pro Ser Gly Ser Asn Gln
35 40 45
Asp Leu Gly Ala Gly Ala Gly Glu Asp Ala Arg Ser Asp Asp Ser Ser
50 55 60
Ser Arg Ile Ile Asn Gly Ser Asp Cys Asp Met His Thr Gln Pro Trp
65 70 75 80
Gln Ala Ala Leu Leu Leu Arg Pro Asn Gln Leu Tyr Cys Gly Ala Val
85 90 95
Leu Val His Pro Gln Trp Leu Leu Thr Ala Ala His Cys Arg Lys Lys
100 105 110
Val Phe Arg Val Arg Leu Gly His Tyr Ser Leu Ser Pro Val Tyr Glu
115 120 125
Ser Gly Gln Gln Met Phe Gln Gly Val Lys Ser Ile Pro His Pro Gly
130 135 140
Tyr Ser His Pro Gly His Ser Asn Asp Leu Met Leu Ile Lys Leu Asn
145 150 155 160
Arg Arg Ile Arg Pro Thr Lys Asp Val Arg Pro Ile Asn Val Ser Ser
165 170 175
His Cys Pro Ser Ala Gly Thr Lys Cys Leu Val Ser Gly Trp Gly Thr
180 185 190
Thr Lys Ser Pro Gln Val His Phe Pro Lys Val Leu Gln Cys Leu Asn
195 200 205
Ile Ser Val Leu Ser Gln Lys Arg Cys Glu Asp Ala Tyr Pro Arg Gln
210 215 220
Ile Asp Asp Thr Met Phe Cys Ala Gly Asp Lys Ala Gly Arg Asp Ser
225 230 235 240
Cys Gln Gly Asp Ser Gly Gly Pro Val Val Cys Asn Gly Ser Leu Gln
245 250 255
Gly Leu Val Ser Trp Gly Asp Tyr Pro Cys Ala Arg Pro Asn Arg Pro
260 265 270
Gly Val Tyr Thr Asn Leu Cys Lys Phe Thr Lys Trp Ile Gln Glu Thr
275 280 285
Ile Gln Ala Asn Ser
290
<210>51
<211>4942
<212>DNA
<213>Homo sapiens
<220>
<223>Corin
<400>51
aaatcatccg tagtgcctcc ccgggggaca cgtagaggag agaaaagcga ccaagataaa 60
agtggacaga agaataagcg agacttttta tccatgaaac agtctcctgc cctcgctccg 120
gaagagcgct gccgcagagc cgggtcccca aagccggtct tgagagctga tgacaataac 180
atgggcaatg gctgctctca gaagctggcg actgctaacc tcctccggtt cctattgctg 240
gtcctgattc catgtatctg tgctctcgtt ctcttgctgg tgatcctgct ttcctatgtt 300
ggaacattac aaaaggtcta ttttaaatca aatgggagtg aacctttggt cactgatggt 360
gaaatccaag ggtccgatgt tattcttaca aatacaattt ataaccagag cactgtggtg 420
tctactgcac atcccgacca acacgttcca gcctggacta cggatgcttc tctcccaggg 480
gaccaaagtc acaggaatac aagtgcctgt atgaacatca cccacagcca gtgtcagatg 540
ctgccctacc acgccacgct gacacctctc ctctcagttg tcagaaacat ggaaatggaa 600
aagttcctca agtttttcac atatctccat cgcctcagtt gctatcaaca tatcatgctg 660
tttggctgta ccctcgcctt ccctgagtgc atcattgatg gcgatgacag tcatggactc 720
ctgccctgta ggtccttctg tgaggctgca aaagaaggct gtgaatcagt cctggggatg 780
gtgaattact cctggccgga tttcctcaga tgctcccagt ttagaaacca aactgaaagc 840
agcaatgtca gcagaatttg cttctcacct cagcaggaaa acggaaagca attgctctgt 900
ggaaggggtg agaactttct gtgtgccagt ggaatctgca tccccgggaa actgcaatgt 960
aatggctaca acgactgtga cgactggagt gacgaggctc attgcaactg cagcgagaat 1020
ctgtttcact gtcacacagg caagtgcctt aattacagcc ttgtgtgtga tggatatgat 1080
gactgtgggg atttgagtga tgagcaaaac tgtgattgca atcccacaac agagcatcgc 1140
tgcggggacg ggcgctgcat cgccatggag tgggtgtgtg atggtgacca cgactgtgtg 1200
gataagtctg acgaggtcaa ctgctcctgt cacagccagg gtctggtgga atgcagaaat 1260
ggacaatgta tccccagcac gtttcaatgt gatggtgacg aggactgcaa ggatgggagt 1320
gatgaggaga actgcagcgt cattcagact tcatgtcaag aaggagacca aagatgcctc 1380
tacaatccct gccttgattc atgtggtggt agctctctct gtgacccgaa caacagtctg 1440
aataactgta gtcaatgtga accaattaca ttggaactct gcatgaattt gccctacaac 1500
agtacaagtt atccaaatta ttttggccac aggactcaaa aggaagcatc catcagctgg 1560
gagtcttctc ttttccctgc acttgttcaa accaactgtt ataaatacct catgttcttt 1620
tcttgcacca ttttggtacc aaaatgtgat gtgaatacag gcgagcatat ccctccttgc 1680
agggcattgt gtgaacactc taaagaacgc tgtgagtctg ttcttgggat tgtgggccta 1740
cagtggcctg aagacacaga ttgcagtcaa tttccagagg aaaattcaga caatcaaacc 1800
tgcctgatgc ctgatgaata tgtggaagaa tgctcaccta gtcatttcaa gtgccgctca 1860
ggacagtgtg ttctggcttc cagaagatgt gatggccagg ccgactgtga cgatgacagt 1920
gatgaggaaa actgtggttg taaagagaga gatctttggg aatgtccatc caataaacaa 1980
tgtttgaagc acacagtgat ctgcgatggg ttcccagact gccctgatta catggacgag 2040
aaaaactgct cattttgcca agatgatgag ctggaatgtg caaaccatgc gtgtgtgtca 2100
cgtgacctgt ggtgtgatgg tgaagccgac tgctcagaca gttcagatga atgggactgt 2160
gtgaccctct ctataaatgt gaactcctct tcctttctga tggttcacag agctgccaca 2220
gaacaccatg tgtgtgcaga tggctggcag gagatattga gtcagctggc ctgcaagcag 2280
atgggtttag gagaaccatc tgtgaccaaa ttgatacagg aacaggagaa agagccgcgg 2340
tggctgacat tacactccaa ctgggagagc ctcaatggga ccactttaca tgaacttcta 2400
gtaaatgggc agtcttgtga gagcagaagt aaaatttctc ttctgtgtac taaacaagac 2460
tgtgggcgcc gccctgctgc ccgaatgaac aaaaggatcc ttggaggtcg gacgagtcgc 2520
cctggaaggt ggccatggca gtgttctctg cagagtgaac ccagtggaca tatctgtggc 2580
tgtgtcctca ttgccaagaa gtgggttctg acagttgccc actgcttcga ggggagagag 2640
aatgctgcag tttggaaagt ggtgcttggc atcaacaatc tagaccatcc atcagtgttc 2700
atgcagacac gctttgtgaa gaccatcatc ctgcatcccc gctacagtcg agcagtggtg 2760
gactatgaca tcagcatcgt tgagctgagt gaagacatca gtgagactgg ctacgtccgg 2820
cctgtctgct tgcccaaccc ggagcagtgg ctagagcctg acacgtactg ctatatcaca 2880
ggctggggcc acatgggcaa taaaatgcca tttaagctgc aagagggaga ggtccgcatt 2940
atttctctgg aacattgtca gtcctacttt gacatgaaga ccatcaccac tcggatgata 3000
tgtgctggct atgagtctgg cacagttgat tcatgcatgg gtgacagcgg tgggcctctt 3060
gtttgtgaga agcctggagg acggtggaca ttatttggat taacttcatg gggctccgtc 3120
tgcttttcca aagtcctggg gcctggcgtt tatagtaatg tgtcatattt cgtcgaatgg 3180
attaaaagac agatttacat ccagaccttt ctcctaaact aattataagg atgatcagag 3240
acttttgcca gctacactaa aagaaaatgg ccttcttgac tgtgaagagc tgcctgcaga 3300
gagctgtaca gaagcacttt tcatggacag aaatgctcaa tcgtgcactg caaatttgca 3360
tgtttgtttt ggactaattt ttttcaattt attttttcac cttcattttt ctcttatttc 3420
aagttcaatg aaagacttta caaaagcaaa caaagcagac tttgtccttt tgccaggcct 3480
aaccatgact gcagcacaaa attatcgact ctggcgagat ttaaaatcag gtgctacagt 3540
aacaggttat ggaatggtct cttttatcct atcacaaaaa aagacataga tatttaggct 3600
gattaattat ctctaccagt ttttgtttct caagctcagt gcatagtggt aaatttcagt 3660
gttaacattg gagacttgct tttctttttc tttttttata ccccacaatt cttttttatt 3720
acacttcgaa ttttagggta cacgagcaca acgtgcaggt tagttacata tgtatacatg 3780
tgccatgttg gtgtgctgaa cccagtaact cgtcatttga tttattaaaa gccaagataa 3840
tttacatgtt taaagtattt actattaccc ccttctaatg tttgcataat tctgagaact 3900
gataaaagac agcaataaaa gaccagtgtc atccatttag gtagcaagac atattgaatg 3960
caaagttctt tagatatcaa tattaacact tgacattatt ggacccccca ttctggatgt 4020
atatcaagat cataatttta tagaagagtc tctatagaac tgtcctcata gctgggtttg 4080
ttcaggatat atgagttggc tgattgagac tgcaacaact acatctatat ttatgggcaa 4140
tattttgttt tacttatgtg gcaaagaact ggatattaaa ctttgcaaaa gagaatttag 4200
atgagagatg caatttttta aaaagaaaat taatttgcat ccctcgttta attaaattta 4260
tttttcagtt ttcttgcgtt catccatacc aacaaagtca taaagagcat attttagagc 4320
acagtaagac tttgcatgga gtaaaacatt ttgtaatttt cctcaaaaga tgtttaatat 4380
ctggtttctt ctcattggta attaaaattt tagaaatgat ttttagctct aggccacttt 4440
acgcaactca atttctgaag caattagtgg taaaaagtat ttttccccac taaaaaactt 4500
taaaacacaa atcttcatat atacttaatt taattagtca ggcatccatt ttgcctttta 4560
aacaactagg attccctact aacctccacc agcaacctgg actgcctcag cattccaaat 4620
agatactacc tgcaatttta tacatgtatt tttgtatctt ttctgtgtgt aaacatagtt 4680
gaaattcaaa aagttgtagc aatttctata ctattcatct cctgtccttc agtttgtata 4740
aacctaagga gagtgtgaaa tccagcaact gaattgtggt cacgattgta tgaaagttca 4800
agaacatatg tcagttttgt tacagttgta gctacatact caatgtatca acttttagcc 4860
tgctcaactt aggctcagtg aaatatatat attatactta ttttaaataa ttcttaatac 4920
aaataaaatg gtaatggtct aa 4942
<210>52
<211>1042
<212>PRT
<213>Homo sapiens
<220>
<223>Corin
<400>52
Met Lys Gln Ser Pro Ala Leu Ala Pro Glu Glu Arg Cys Arg Arg Ala
1 5 10 15
Gly Ser Pro Lys Pro Val Leu Arg Ala Asp Asp Asn Asn Met Gly Asn
20 25 30
Gly Cys Ser Gln Lys Leu Ala Thr Ala Asn Leu Leu Arg Phe Leu Leu
35 40 45
Leu Val Leu Ile Pro Cys Ile Cys Ala Leu Val Leu Leu Leu Val Ile
50 55 60
Leu Leu Ser Tyr Val Gly Thr Leu Gln Lys Val Tyr Phe Lys Ser Asn
65 70 75 80
Gly Ser Glu Pro Leu Val Thr Asp Gly Glu Ile Gln Gly Ser Asp Val
85 90 95
Ile Leu Thr Asn Thr Ile Tyr Asn Gln Ser Thr Val Val Ser Thr Ala
100 105 110
His Pro Asp Gln His Val Pro Ala Trp Thr Thr Asp Ala Ser Leu Pro
115 120 125
Gly Asp Gln Ser His Arg Asn Thr Ser Ala Cys Met Asn Ile Thr His
130 135 140
Ser Gln Cys Gln Met Leu Pro Tyr His Ala Thr Leu Thr Pro Leu Leu
145 150 155 160
Ser Val Val Arg Asn Met Glu Met Glu Lys Phe Leu Lys Phe Phe Thr
165 170 175
Tyr Leu His Arg Leu Ser Cys Tyr Gln His Ile Met Leu Phe Gly Cys
180 185 190
Thr Leu Ala Phe Pro Glu Cys Ile Ile Asp Gly Asp Asp Ser His Gly
195 200 205
Leu Leu Pro Cys Arg Ser Phe Cys Glu Ala Ala Lys Glu Gly Cys Glu
210 215 220
Ser Val Leu Gly Met Val Asn Tyr Ser Trp Pro Asp Phe Leu Arg Cys
225 230 235 240
Ser Gln Phe Arg Asn Gln Thr Glu Ser Ser Asn Val Ser Arg Ile Cys
245 250 255
Phe Ser Pro Gln Gln Glu Asn Gly Lys Gln Leu Leu Cys Gly Arg Gly
260 265 270
Glu Asn Phe Leu Cys Ala Ser Gly Ile Cys Ile Pro Gly Lys Leu Gln
275 280 285
Cys Asn Gly Tyr Asn Asp Cys Asp Asp Trp Ser Asp Glu Ala His Cys
290 295 300
Asn Cys Ser Glu Asn Leu Phe His Cys His Thr Gly Lys Cys Leu Asn
305 310 315 320
Tyr Ser Leu Val Cys Asp Gly Tyr Asp Asp Cys Gly Asp Leu Ser Asp
325 330 335
Glu Gln Asn Cys Asp Cys Asn Pro Thr Thr Glu His Arg Cys Gly Asp
340 345 350
Gly Arg Cys Ile Ala Met Glu Trp Val Cys Asp Gly Asp His Asp Cys
355 360 365
Val Asp Lys Ser Asp Glu Val Asn Cys Ser Cys His Ser Gln Gly Leu
370 375 380
Val Glu Cys Arg Asn Gly Gln Cys Ile Pro Ser Thr Phe Gln Cys Asp
385 390 395 400
Gly Asp Glu Asp Cys Lys Asp Gly Ser Asp Glu Glu Asn Cys Ser Val
405 410 415
Ile Gln Thr Ser Cys Gln Glu Gly Asp Gln Arg Cys Leu Tyr Asn Pro
420 425 430
Cys Leu Asp Ser Cys Gly Gly Ser Ser Leu Cys Asp Pro Asn Asn Ser
435 440 445
Leu Asn Asn Cys Ser Gln Cys Glu Pro Ile Thr Leu Glu Leu Cys Met
450 455 460
Asn Leu Pro Tyr Asn Ser Thr Ser Tyr Pro Asn Tyr Phe Gly His Arg
465 470 475 480
Thr Gln Lys Glu Ala Ser Ile Ser Trp Glu Ser Ser Leu Phe Pro Ala
485 490 495
Leu Val Gln Thr Asn Cys Tyr Lys Tyr Leu Met Phe Phe Ser Cys Thr
500 505 510
Ile Leu Val Pro Lys Cys Asp Val Asn Thr Gly Glu His Ile Pro Pro
515 520 525
Cys Arg Ala Leu Cys Glu His Ser Lys Glu Arg Cys Glu Ser Val Leu
530 535 540
Gly Ile Val Gly Leu Gln Trp Pro Glu Asp Thr Asp Cys Ser Gln Phe
545 550 555 560
Pro Glu Glu Asn Ser Asp Asn Gln Thr Cys Leu Met Pro Asp Glu Tyr
565 570 575
ValGlu Glu Cys Ser Pro Ser His Phe Lys Cys Arg Ser Gly Gln Cys
580 585 590
Val Leu Ala Ser Arg Arg Cys Asp Gly Gln Ala Asp Cys Asp Asp Asp
595 600 605
Ser Asp Glu Glu Asn Cys Gly Cys Lys Glu Arg Asp Leu Trp Glu Cys
610 615 620
Pro Ser Asn Lys Gln Cys Leu Lys His Thr Val Ile Cys Asp Gly Phe
625 630 635 640
Pro Asp Cys Pro Asp Tyr Met Asp Glu Lys Asn Cys Ser Phe Cys Gln
645 650 655
Asp Asp Glu Leu Glu Cys Ala Asn His Ala Cys Val Ser Arg Asp Leu
660 665 670
Trp Cys Asp Gly Glu Ala Asp Cys Ser Asp Ser Ser Asp Glu Trp Asp
675 680 685
Cys Val Thr Leu Ser Ile Asn Val Asn Ser Ser Ser Phe Leu Met Val
690 695 700
His Arg Ala Ala Thr Glu His His ValCys Ala Asp Gly Trp Gln Glu
705 710 715 720
Ile Leu Ser Gln Leu Ala Cys Lys Gln Met Gly Leu Gly Glu Pro Ser
725 7307 35
Val Thr Lys Leu Ile Gln Glu Gln Glu Lys Glu Pro Arg Trp Leu Thr
740 745 750
Leu His Ser Asn Trp Glu Ser Leu Asn Gly Thr Thr Leu His Glu Leu
755 760 765
Leu Val Asn Gly Gln Ser Cys Glu Ser Arg Ser Lys Ile Ser Leu Leu
770 775 780
Cys Thr Lys Gln Asp Cys Gly Arg Arg Pro Ala Ala Arg Met Asn Lys
785 790 795 800
Arg Ile Leu Gly Gly Arg Thr Ser Arg Pro Gly Arg Trp Pro Trp Gln
805 810 815
Cys Ser Leu Gln Ser Glu Pro Ser Gly His Ile Cys Gly Cys Val Leu
820 825 830
Ile Ala Lys Lys Trp Val Leu Thr Val Ala His Cys Phe Glu Gly Arg
835 840 845
Glu Asn Ala Ala Val Trp Lys Val Val Leu Gly Ile Asn Asn Leu Asp
850 855 860
His Pro Ser Val Phe Met Gln Thr Arg Phe Val Lys Thr Ile Ile Leu
865 870 875 880
His Pro Arg Tyr Ser Arg Ala Val Val Asp Tyr Asp Ile Ser Ile Val
885 890 895
Glu Leu Ser Glu Asp Ile Ser Glu Thr Gly Tyr Val Arg Pro Val Cys
900 905 910
Leu Pro Asn Pro Glu Gln Trp Leu Glu Pro Asp Thr Tyr Cys Tyr Ile
915 920 925
Thr Gly Trp Gly His Met Gly Asn Lys Met Pro Phe Lys Leu Gln Glu
930 935 940
Gly Glu Val Arg Ile Ile Ser Leu Glu His Cys Gln Ser Tyr Phe Asp
945 950 955 960
Met Lys Thr Ile Thr Thr Arg Met Ile Cys Ala Gly Tyr Glu Ser Gly
965 970 975
Thr Val Asp Ser Cys Met Gly Asp Ser Gly Gly Pro Leu Val Cys Glu
980 985 990
Lys Pro Gly Gly Arg Trp Thr Leu Phe Gly Leu Thr Ser Trp Gly Ser
995 1000 1005
Val Cys Phe Ser Lys Val Leu Gly Pro Gly Val Tyr Ser Asn Val Ser
1010 1015 1020
Tyr Phe Val Glu Trp Ile Lys Arg Gln Ile Tyr Ile Gln Thr Phe Leu
1025 1030 1035 1040
Leu Asn
<210>53
<211>945
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein 12 transcript variant 1
<400>53
gcaggtaggt ggacggagag atagcagcga cgaggacagg ccaaacagtg acagccacgt 60
agaggatctg gcagacaaag agacaaggtg agaaggagac tttggaagtg acccaccatg 120
gggctcagca tctttttgct cctgtgtgtt cttgggctca gccaggcagc cacaccgaag 180
attttcaatg gcactgagtg tgggcgtaac tcacagccgt ggcaggtggg gctgtttgag 240
ggcaccagcc tgcgctgcgg gggtgtcctt attgaccaca ggtgggtcct cacagcggct 300
cactgcagcg gcagcaggta ctgggtgcgc ctgggggaac acagcctcag ccagctcgac 360
tggaccgagc agatccggca cagcggcttc tctgtgaccc atcccggcta cctgggagcc 420
tcgacgagcc acgagcacga cctccggctg ctgcggctge gcctgcccgt ccgcgtaacc 480
agcagcgttc aacccctgcc cctgcccaat gactgtgcaa ccgctggcac cgagtgccac 540
gtctcaggct ggggcatcac caaccaccca cggaacccat tcccggatct gctccagtgc 600
ctcaacctct ccatcgtctc ccatgccacc tgccatggtg tgtatcccgg gagaatcacg 660
agcaacatgg tgtgtgcagg cggcgtcccg gggcaggatg cctgccaggg tgattctggg 720
ggccccctgg tgtgtggggg agtccttcaa ggtctggtgt cctgggggtc tgtggggccc 780
tgtggacaag atggcatccc tggagtctac acctatattt gcaactccac tcttgttggc 840
ctgggaactt cttggaactt taactcctgc cagcccttct aagacccacg agcggggtga 900
gagaagtgtg caatagtctg gaataaatat aaatgaagga ggggc 945
<210>54
<211>254
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein 12 isoform 1
<400>54
Met Gly Leu Ser Ile Phe Leu Leu Leu Cys Val Leu Gly Leu Ser Gln
1 5 10 15
Ala Ala Thr Pro Lys Ile Phe Asn Gly Thr Glu Cys Gly Arg Asn Ser
20 25 30
Gln Pro Trp Gln Val Gly Leu Phe Glu Gly Thr Ser Leu Arg Cys Gly
35 40 45
Gly Val Leu Ile Asp His Arg Trp Val Leu Thr Ala Ala His Cys Ser
50 55 60
Gly Ser Arg Tyr Trp Val Arg Leu Gly Glu His Ser Leu Ser Gln Leu
65 70 75 80
Asp Trp Thr Glu Gln Ile Arg His Ser Gly Phe Ser Val Thr His Pro
85 90 95
Gly Tyr Leu Gly Ala Ser Thr Ser His Glu His Asp Leu Arg Leu Leu
100 105 110
Arg Leu Arg Leu Pro Val Arg Val Thr Ser Ser Val Gln Pro Leu Pro
115 120 125
Leu Pro Asn Asp Cys Ala Thr Ala Gly Thr Glu Cys His Val Ser Gly
130 135 140
Trp Gly Ile Thr Asn His Pro Arg Asn Pro Phe Pro Asp Leu Leu Gln
145 150 155 160
Cys Leu Asn Leu Ser Ile Val Ser His Ala Thr Cys His Gly Val Tyr
165 170 175
Pro Gly Arg Ile Thr Ser Asn Met Val Cys Ala Gly Gly Val Pro Gly
180 185 190
Gln Asp Ala Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Gly Gly
195 200 205
Val Leu Gln Gly Leu Val Ser Trp Gly Ser Val Gly Pro Cys Gly Gln
210 215 220
Asp Gly Ile Pro Gly Val Tyr Thr Tyr Ile Cys Asn Ser Thr Leu Val
225 230 235 240
Gly Leu Gly Thr Ser Trp Asn Phe Asn Ser Cys Gln Pro Phe
245 250
<210>55
<211>1074
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein 12 transcript variant 2
<400>55
gcaggtaggt ggacggagag atagcagcga cgaggacagg ccaaacagtg acagccacgt 60
agaggatctg gcagacaaag agacaaggtg agaaggagac tttggaagtg acccaccatg 120
gggctcagca tctttttgct cctgtgtgtt cttgggctca gccaggcagc cacaccgaag 180
attttcaatg gcactgagtg tgggcgtaac tcacagccgt ggcaggtggg gctgtttgag 240
ggcaccagcc tgcgctgcgg gggtgtcctt attgaccaca ggtgggtcct cacagcggct 300
cactgcagcg gcagcaggta ctgggtgcgc ctgggggaac acagcctcag ccagctcgac 360
tggaccgagc agatccggca cagcggcttc tctgtgaccc atcccggcta cctgggagcc 420
tcgacgagcc acgagcacga cctccggctg ctgcggctgc gcctgcccgt ccgcgtaacc 480
agcagcgttc aacccctgcc cctgcccaat gactgtgcaa ccgctggcac cgagtgccac 540
gtctcaggct ggggcatcac caaccaccca cggaacccat tcccggatct gctccagtgc 600
ctcaacctct ccatcgtctc ccatgccacc tgccatggtg tgtatcccgg gagaatcacg 660
agcaacatgg tgtgtgcagg cggcgtcccg gggcaggatg cctgccaggg tgattctggg 720
ggccccctgg tgtgtggggg agtccttcaa ggtctggtgt cctgggggtc tgtggggccc 780
tgtggacaag atggcatccc tggagtctac acctatattt gcaagtatgt ggactggatc 840
cggatgatca tgaggaacaa ctgacctgtt tcctccacct ccacccccac cccttaactt 900
gggtacccct ctggccctca gagcaccaat atctcctcca tcacttcccc tagctccact 960
cttgttggcc tgggaacttc ttggaacttt aactcctgcc agcccttcta agacccacga 1020
gcggggtgag agaagtgtgc aatagtctgg aataaatata aatgaaggag gggc 1074
<210>56
<211>248
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein 12 isoform 2
<400>56
Met Gly Leu Ser Ile Phe Leu Leu Leu Cys Val Leu Gly Leu Ser Gln
1 5 10 15
Ala Ala Thr Pro Lys Ile Phe Asn Gly Thr Glu Cys Gly Arg Asn Ser
20 25 30
Gln Pro Trp Gln Val Gly Leu Phe Glu Gly Thr Ser Leu Arg Cys Gly
35 40 45
Gly Val Leu Ile Asp His Arg Trp Val Leu Thr Ala Ala His Cys Ser
50 55 60
Gly Ser Arg Tyr Trp Val Arg Leu Gly Glu His Ser Leu Ser Gln Leu
65 70 75 80
Asp Trp Thr Glu Gln Ile Arg His Ser Gly Phe Ser Val Thr His Pro
85 90 95
Gly Tyr Leu Gly Ala Ser Thr Ser His Glu His Asp Leu Arg Leu Leu
100 105 110
Arg Leu Arg Leu Pro Val Arg Val Thr Ser Ser Val Gln Pro Leu Pro
115 120 125
Leu Pro Asn Asp Cys Ala Thr Ala Gly Thr Glu Cys His Val Ser Gly
130 135 140
Trp Gly Ile Thr Asn His Pro Arg Asn Pro Phe Pro Asp Leu Leu Gln
145 150 155 160
Cys Leu Asn Leu Ser Ile Val Ser His Ala Thr Cys His Gly Val Tyr
165 170 175
Pro Gly Arg Ile Thr Ser Asn Met Val Cys Ala Gly Gly Val Pro Gly
180 185 190
Gln Asp Ala Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Gly Gly
195 200 205
Val Leu Gln Gly Leu Val Ser Trp Gly Ser ValGly Pro Cys Gly Gln
210 215 220
Asp Gly Ile Pro Gly Val Tyr Thr Tyr Ile Cys Lys Tyr Val Asp Trp
225 230 235 240
Ile Arg Met Ile Met Arg Asn Asn
245
<210>57
<211>814
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein 12 transcript variant 3
<400>57
gcaggtaggt ggacggagag atagcagcga cgaggacagg ccaaacagtg acagccacgt 60
agaggatctg gcagacaaag agacaaggtg agaaggagac tttggaagtg acccaccatg 120
gggctcagca tctttttgct cctgtgtgtt cttgggctca gccaggcagc cacaccgaag 180
attttcaatg gcactgagtg tgggcgtaac tcacagccgt ggcaggtggg gctgtttgag 240
ggcaccagcc tgcgctgcgg gggtgtcctt attgaccaca ggtgggtcct cacagcggct 300
cactgcagcg gcagacccat tcccggatct gctccagtgc ctcaacctct ccatcgtctc 360
ccatgccacc tgccatggtg tgtatcccgg gagaatcacg agcaacatgg tgtgtgcagg 420
cggcgtcccg gggcaggatg cctgccaggg tgattctggg ggccccctgg tgtgtggggg 480
agtccttcaa ggtctggtgt cctgggggtc tgtggggccc tgtggacaag atggcatccc 540
tggagtctac acctatattt gcaagtatgt ggactggatc cggatgatca tgaggaacaa 600
ctgacctgtt tcctccacct ccacccccac cccttaactt gggtacccct ctggccctca 660
gagcaccaat atctcctcca tcacttcccc tagctccact cttgttggcc tgggaacttc 720
ttggaacttt aactcctgcc agcccttcta agacccacga gcggggtgag agaagtgtgc 780
aatagtctgg aataaatata aatgaaggag gggc 814
<210>58
<211>111
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein 12 isoform 3
<400>58
Met Gly Leu Ser Ile Phe Leu Leu Leu Cys Val Leu Gly Leu Ser Gln
1 5 10 15
Ala Ala Thr Pro Lys Ile Phe Asn Gly Thr Glu Cys Gly Arg Asn Ser
20 25 30
Gln Pro Trp Gln Val Gly Leu Phe Glu Gly Thr Ser Leu Arg Cys Gly
35 40 45
Gly Val Leu Ile Asp His Arg Trp Val Leu Thr Ala Ala His Cys Ser
50 55 60
Gly Arg Pro Ile Pro Gly Ser Ala Pro Val Pro Gln Pro Leu His Arg
65 70 75 80
Leu Pro Cys His Leu Pro Trp Cys Val Ser Arg Glu Asn His Glu Gln
85 90 95
His Gly Val Cys Arg Arg Arg Pro Gly Ala Gly Cys Leu Pro Gly
100 105 110
<210>59
<211>1471
<212>DNA
<213>Homo sapiens
<220>
<223>DESC1 oritease
<400>59
tgacttggat gtagacctcg accttcacag gactcttcat tgctggttgg caatgatgta 60
tcggccagat gtggtgaggg ctaggaaaag agtttgttgg gaaccctggg ttatcggcct 120
cgtcatcttc atatccctga ttgtcctggc agtgtgcatt ggactcactg ttcattatgt 180
gagatataat caaaagaaga cctacaatta ctatagcaca ttgtcattta caactgacaa 240
actatatgct gagtttggca gagaggcttc taacaatttt acagaaatga gccagagact 300
tgaatcaatg gtgaaaaatg cattttataa atctccatta agggaagaat ttgtcaagtc 360
tcaggttatc aagttcagtc aacagaagca tggagtgttg gctcatatgc tgttgatttg 420
tagatttcac tctactgagg atcctgaaac tgtagataaa attgttcaac ttgttttaca 480
tgaaaagctg caagatgctg taggaccccc taaagtagat cctcactcag ttaaaattaa 540
aaaaatcaac aagacagaaa cagacagcta tctaaaccat tgctgcggaa cacgaagaag 600
taaaactcta ggtcagagtc tcaggatcgt tggtgggaca gaagtagaag agggtgaatg 660
gccctggcag gctagcctgc agtgggatgg gagtcatcgc tgtggagcaa ccttaattaa 720
tgccacatgg cttgtgagtg ctgctcactg ttttacaaca tataagaacc ctgccagatg 780
gactgcttcc tttggagtaa caataaaacc ttcgaaaatg aaacggggtc tccggagaat 840
aattgtccat gaaaaataca aacacccatc acatgactat gatatttctc ttgcagagct 900
ttctagccct gttccctaca caaatgcagt acatagagtt tgtctccctg atgcatccta 960
tgagtttcaa ccaggtgatg tgatgtttgt gacaggattt ggagcactga aaaatgatgg 1020
ttacagtcaa aatcatcttc gacaagcaca ggtgactctc atagacgcta caacttgcaa 1080
tgaacctcaa gcttacaatg acgccataac tcctagaatg ttatgtgctg gctccttaga 1140
aggaaaaaca gatgcatgcc agggtgactc tggaggacca ctggttagtt cagatgctag 1200
agatatctgg taccttgctg gaatagtgag ctggggagat gaatgtgcga aacccaacaa 1260
gcctggtgtt tatactagag ttacggcctt gcgggactgg attacttcaa aaactggtat 1320
ctaagagaga aaagcctcat ggaacagata acattttttt ttgttttttg ggtgtggagg 1380
ccatttttag agatacagaa ttggagaaga cttgcaaaac agctagattt gactgatctc 1440
aataaactgt ttgcttgatg caaaaaaaaa a 1471
<210>60
<211>422
<212>PRT
<213>Homo sapiens
<220>
<223>DESC1 oritease
<400>60
Met Tyr Arg Pro Asp Val Val Arg Ala Arg Lys Arg Val Cys Trp Glu
1 5 10 15
Pro Trp Val Ile Gly Leu Val Ile Phe Ile Ser Leu Ile Val Leu Ala
20 25 30
Val Cys Ile Gly Leu Thr Val His Tyr Val Arg Tyr Asn Gln Lys Lys
35 40 45
Thr Tyr Asn Tyr Tyr Ser Thr Leu Ser Phe Thr Thr Asp Lys Leu Tyr
50 55 60
Ala Glu Phe Gly Arg Glu Ala Ser Asn Asn Phe Thr Glu Met Ser Gln
65 70 75 80
Arg Leu Glu Ser Met Val Lys Asn Ala Phe Tyr Lys Ser Pro Leu Arg
85 90 95
Glu Glu Phe Val Lys Ser Gln Val Ile Lys Phe Ser Gln Gln Lys His
100 105 110
Gly Val Leu Ala His Met Leu Leu Ile Cys Arg Phe His Ser Thr Glu
115 120 125
Asp Pro Glu Thr Val Asp Lys Ile Val Gln Leu Val Leu His Glu Lys
130 135 140
Leu Gln Asp Ala Val Gly Pro Pro Lys Val Asp Pro His Ser Val Lys
145 150 155 160
Ile Lys Lys Ile Asn Lys Thr Glu Thr Asp Ser Tyr Leu Asn His Cys
165 170 175
Cys Gly Thr Arg Arg Ser Lys Thr Leu Gly Gln Ser Leu Arg Ile Val
180 185 190
Gly Gly Thr Glu Val Glu Glu Gly Glu Trp Pro Trp Gln Ala Ser Leu
195 200 205
Gln Trp Asp Gly Ser His Arg Cys Gly Ala Thr Leu Ile Asn Ala Thr
210 215 220
Trp Leu Val Ser Ala Ala His Cys Phe Thr Thr Tyr Lys Asn Pro Ala
225 230 235 240
Arg Trp Thr Ala Ser Phe Gly Val Thr Ile Lys Pro Ser Lys Met Lys
245 250 255
Arg Gly Leu Arg Arg Ile Ile Val His Glu Lys Tyr Lys His Pro Ser
260 265 270
His Asp Tyr Asp Ile Ser Leu Ala Glu Leu Ser Ser Pro Val Pro Tyr
275 280 285
Thr Asn Ala Val His Arg Val Cys Leu Pro Asp Ala Ser Tyr Glu Phe
290 295 300
Gln Pro Gly Asp Val Met Phe Val Thr Gly Phe Gly Ala Leu Lys Asn
305 310 315 320
Asp Gly Tyr Ser Gln Asn His Leu Arg Gln Ala Gln Val Thr Leu Ile
325 330 335
Asp Ala Thr Thr Cys Asn Glu Pro Gln Ala Tyr Asn Asp Ala Ile Thr
340 345 350
Pro Arg Met Leu Cys Ala Gly Ser Leu Glu Gly Lys Thr Asp Ala Cys
355 360 365
Gln Gly Asp Ser Gly Gly Pro Leu Val Ser Ser Asp Ala Arg Asp Ile
370 375 380
Trp Tyr Leu Ala Gly Ile Val Ser Trp Gly Asp Glu Cys Ala Lys Pro
385 390 395 400
Asn Lys Pro Gly Val Tyr Thr Arg Val Thr Ala Leu Arg Asp Trp Ile
405 410 415
Thr Ser Lys Thr Gly Ile
420
<210>61
<211>1124
<212>DNA
<213>Homo sapiens
<220>
<223>tryptase gamma 1
<400>61
ccatggccct tggggcctgt ggcctcctgc tgctcctggc tgtgcccggt gtgtccctca 60
ggactttgca gccagggtgt ggccggccgc aggtttcgga tgcaggcggc cggatcgtgg 120
ggggtcacgc tgccccggcc ggcgcatggc catggcaggc cagcctccgc ctgcggagga 180
tgcacgtgtg cggcgggtca ctgctcagcc cccagtgggt gctcacagct gcccactgct 240
tctccgggtc cctgaactca tccgactacc aggtgcacct gggggaactg gagatcactt 300
tgtctcccca cttctccacc gtgaggcaga tcatcctgca ctccagcccc tcaggacagc 360
cggggaccag cggggacatc gccctggtgg agctcagtgt ccccgtgacc ctctccagcc 420
ggatcctgcc cgtctgcctc ccggaggcct cagatgactt ctgccctggg atccggtgct 480
gggtgaccgg ctggggctat acgcgggagg gagagcctct gccacccccg tacagcctgc 540
gggaggtgaa agtctccgtg gtggacacag agacctgccg ccgggactat cccggccccg 600
ggggcagcat ccttcagccc gacatgctgt gtgcccgggg ccccggggat gcctgccagg 660
acgactccgg ggggcctctg gtctgccagg tgaacggtgc ctgggtgcag gctggcattg 720
tgagctgggg tgagggctgc ggccgcccca acaggccggg agtctacact cgtgtccctg 780
cctacgtgaa ctggatccgc cgccacatca cagcatcagg gggctcagag tctgggtacc 840
ccaggctccc cctcctggct ggcttcttcc tccccggcct cttccttctg ctagtctcct 900
gtgtcctgct ggccaagtgc ctgctgcacc catctgcgga tggtactccc ttccccgccc 960
ctgactgatg gcaggaatcc aagtgcattt cttaaataag ttactattta ttccgctccg 1020
ccccctccct ctcccttgag aagctgagtc ttctgcatca gattattgca acatttaacc 1080
tgaatttaaa agcacacgaa acaaaaaaaa aaaaaaaaaa aaaa 1124
<210>62
<211>321
<212>PRT
<213>Homo sapiens
<220>
<223>tryptase gamma 1
<400>62
Met Ala Leu Gly Ala Cys Gly Leu Leu Leu Leu Leu Ala Val Pro Gly
1 5 10 15
Val Ser Leu Arg Thr Leu Gln Pro Gly Cys Gly Arg Pro Gln Val Ser
20 25 30
Asp Ala Gly Gly Arg Ile Val Gly Gly His Ala Ala Pro Ala Gly Ala
35 40 45
Trp Pro Trp Gln Ala Ser Leu Arg Leu Arg Arg Met His Val Cys Gly
50 55 60
Gly Ser Leu Leu Ser Pro Gln Trp Val Leu Thr Ala Ala His Cys Phe
65 70 75 80
Ser Gly Ser Leu Asn Ser Ser Asp Tyr Gln Val His Leu Gly Glu Leu
85 90 95
Glu Ile Thr Leu Ser Pro His Phe Ser Thr Val Arg Gln Ile Ile Leu
100 105 110
His Ser Ser Pro Ser Gly Gln Pro Gly Thr Ser Gly Asp Ile Ala Leu
115 120 125
Val Glu Leu Ser Val Pro Val Thr Leu Ser Ser Arg Ile Leu Pro Val
130 135 140
Cys Leu Pro Glu Ala Ser Asp Asp Phe Cys Pro Gly Ile Arg Cys Trp
145 150 155 160
Val Thr Gly Trp Gly Tyr Thr Arg Glu Gly Glu Pro Leu Pro Pro Pro
165 170 175
Tyr Ser Leu Arg Glu Val Lys Val Ser Val Val Asp Thr Glu Thr Cys
180 185 190
Arg Arg Asp Tyr Pro Gly Pro Gly Gly Ser Ile Leu Gln Pro Asp Met
195 200 205
Leu Cys Ala Arg Gly Pro Gly Asp Ala Cys Gln Asp Asp Ser Gly Gly
210 215 220
Pro Leu Val Cys Gln Val Asn Gly Ala Trp Val Gln Ala Gly Ile Val
225 230 235 240
Ser Trp Gly Glu Gly Cys Gly Arg Pro Asn Arg Pro Gly Val Tyr Thr
245 250 255
Arg Val Pro Ala Tyr Val Asn Trp Ile Arg Arg His Ile Thr Ala Ser
260 265 270
Gly Gly Ser Glu Ser Gly Tyr Pro Arg Leu Pro Leu Leu Ala Gly Phe
275 280 285
Phe Leu Pro Gly Leu Phe Leu Leu Leu Val Ser Cys Val Leu Leu Ala
290 295 300
Lys Cys Leu Leu His Pro Ser Ala Asp Gly Thr Pro Phe Pro Ala Pro
305 310 315 320
Asp
<210>63
<211>1459
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK14
<400>63
gcacctagca gatcccaagc tggggaatgc cagttctggc accaaccttc ctgctccctg 60
ctggggcctc tgctccccca tctctcagga gtcgaaagtg agaaagcaag acatcaagga 120
gggacctgtg ccctgctcca catcctccca cctgccgccc gcagagcctg caggccccgc 180
ccccctcgtc tctggtccct acctctctgc tgtgtcttca tgtccctgag ggtcttgggc 240
tctgggacct ggccctcagc ccctaaaatg ttcctcctgc tgacagcact tcaagtcctg 300
gctatagcca tgacacagag ccaagaggat gagaacaaga taattggtgg ccatacgtgc 360
acccggagct cccagccgtg gcaggcggcc ctgctggcgg gtcccaggcg ccgcttcctc 420
tgcggaggcg ccctgctttc aggccagtgg gtcatcactg ctgctcactg cggccgcccg 480
atccttcagg ttgccctggg caagcacaac ctgaggaggt gggaggccac ccagcaggtg 540
ctgcgcgtgg ttcgtcaggt gacgcacccc aactacaact cccggaccca cgacaacgac 600
ctcatgctgc tgcagctaca gcagcccgca cggatcggga gggcagtcag gcccattgag 660
gtcacccagg cctgtgccag ccccgggacc tcctgccgag tgtcaggctg gggaactata 720
tccagcccca tcgccaggta ccccgcctct ctgcaatgcg tgaacatcaa catctccccg 780
gatgaggtgt gccagaaggc ctatcctaga accatcacgc ctggcatggt ctgtgcagga 840
gttccccagg gcgggaagga ctcttgtcag ggtgactctg ggggacccct ggtgtgcaga 900
ggacagctcc agggcctcgt gtcttgggga atggagcgct gcgccctgcc tggctacccc 960
ggtgtctaca ccaacctgtg caagtacaga agctggattg aggaaacgat gcgggacaaa 1020
tgatggtctt cacggtggga tggacctcgt cagctgccca ggccctcctc tctctactca 1080
ggacccagga gtccaggccc cagcccctcc tccctcagac ccaggagtcc aggcccccag 1140
cccctcctcc ctcagacccg ggagtccagg cccccagccc ctcctccctc agacccagga 1200
gccccagtcc ccagcatcct gatctttact ccggctctga tctctccttt cccagagcag 1260
ttgcttcagg cgttttctcc ccaccaagcc cccacccttg ctgtgtcacc atcactactc 1320
aagaccggag gcacagaggg caggagcaca gaccccttaa accggcattg tattccaaag 1380
acgacaattt ttaacacgct tagtgtctct aaaaaccgaa taaataatga caataaaaat 1440
ggaatcatcc taaattgta 1459
<210>64
<211>251
<212>PRT
<213>Homo sapiens
<220>
<223>Kallikrein-14 precursor S01.029
<400>64
Met Phe Leu Leu Leu Thr Ala Leu Gln Val Leu Ala Ile Ala Met Thr
1 5 10 15
Gln Ser Gln Glu Asp Glu Asn Lys Ile Ile Gly Gly His Thr Cys Thr
20 25 30
Arg Ser Ser Gln Pro Trp Gln Ala Ala Leu Leu Ala Gly Pro Arg Arg
35 40 45
Arg Phe Leu Cys Gly Gly Ala Leu Leu Ser Gly Gln Trp Val Ile Thr
50 55 60
Ala Ala His Cys Gly Arg Pro Ile Leu Gln Val Ala Leu Gly Lys His
65 70 75 80
Asn Leu Arg Arg Trp Glu Ala Thr Gln Gln Val Leu Arg Val Val Arg
85 90 95
Gln Val Thr His Pro Asn Tyr Asn Ser Arg Thr His Asp Asn Asp Leu
100 105 110
Met Leu Leu Gln Leu Gln Gln Pro Ala Arg Ile Gly Arg Ala Val Arg
115 120 125
Pro Ile Glu Val Thr Gln Ala Cys Ala Ser Pro Gly Thr Ser Cys Arg
130 135 140
Val Ser Gly Trp Gly Thr Ile Ser Ser Pro Ile Ala Arg Tyr Pro Ala
145 150 155 160
Ser Leu Gln Cys Val Asn Ile Asn Ile Ser Pro Asp Glu Val Cys Gln
165 170 175
Lys Ala Tyr Pro Arg Thr Ile Thr Pro Gly Met Val Cys Ala Gly Val
180 185 190
Pro Gln Gly Gly Lys Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu
195 200 205
Val Cys Arg Gly Gln Leu Gln Gly Leu Val Ser Trp Gly Met Glu Arg
210 215 220
Cys Ala Leu Pro Gly Tyr Pro Gly Val Tyr Thr Asn Leu Cys Lys Tyr
225 230 235 240
Arg Ser Trp Ile Glu Glu Thr Met Arg Asp Lys
245 250
<210>65
<211>3008
<212>DNA
<213>Homo sapiens
<220>
<223>hyaluronan-binding serine protease(HGF activator-like protein)
<400>65
cctgaatcct tggagactga catttttccc ccctaaaggc atagacaaca aaagaaattt 60
tattgagagg aaaacacaag tccttaaact gcaaagatgt ttgccaggat gtctgatctc 120
catgttctgc tgttaatggc tctggtggga aagacagcct gtgggttctc cctgatgtct 180
ttattggaaa gcctggaccc agactggacc cctgaccagt atgattacag ctacgaggat 240
tataatcagg aagagaacac cagtagcaca cttacccacg ctgagaatcc tgactggtac 300
tacactgagg accaagctga tccatgccag cccaacccct gtgaacacgg tggggactgc 360
ctcgtccatg ggagcacctt cacatgcagc tgcctggctc ctttctctgg gaataagtgt 420
cagaaagtgc aaaatacgtg caaggacaac ccatgtggcc ggggccaatg tctcattacc 480
cagagtcctc cctactaccg ctgtgtctgt aaacaccctt acacaggtcc cagctgctcc 540
caagtggttc ctgtatgcag gccaaacccc tgccagaatg gggctacctg ctcccggcat 600
aagcggagat ccaagttcac ctgtgcctgt cccgaccagt tcaaggggaa attctgtgaa 660
ataggttctg atgactgcta tgttggcgat ggctactctt accgagggaa aatgaatagg 720
acagtcaacc agcatgcgtg cctttactgg aactcccacc tcctcttgca ggagaattac 780
aacatgttta tggaggatgc tgaaacccat gggattgggg aacacaattt ctgcagaaac 840
ccagatgcgg acgaaaagcc ctggtgcttt attaaagtta ccaatgacaa ggtgaaatgg 900
gaatactgtg atgtctcagc ctgctcagcc caggacgttg cctacccaga ggaaagcccc 960
actgagccat caaccaagct tccggggttt gactcctgtg gaaagactga gatagcagag 1020
aggaagatca agagaatcta tggaggcttt aagagcacgg cgggcaagca cccatggcag 1080
gcgtccctcc agtcctcgct gcctctgacc atctccatgc cccagggcca cttctgtggt 1140
ggggcgctga tccacccctg ctgggtgctc actgctgccc actgcaccga cataaaaacc 1200
agacatctaa aggtggtgct aggggaccag gacctgaaga aagaagaatt tcatgagcag 1260
agctttaggg tggagaagat attcaagtac agccactaca atgaaagaga tgagattccc 1320
cacaatgata ttgcattgct caagttaaag ccagtggatg gtcactgtgc tctagaatcc 1380
aaatacgtga agactgtgtg cttgcctgat gggtcctttc cctctgggag tgagtgccac 1440
atctctggct ggggtgttac agaaacagga aaagggtccc gccagctcct ggatgccaaa 1500
gtcaagctga ttgccaacac tttgtgcaac tcccgccaac tctatgacca catgattgat 1560
gacagtatga tctgtgcagg aaatcttcag aaacctgggc aagacacctg ccagggtgac 1620
tctggaggcc ccctgacctg tgagaaggac ggcacctact acgtctatgg gatagtgagc 1680
tggggcctgg agtgtgggaa gaggccaggg gtctacaccc aagttaccaa attcctgaat 1740
tggatcaaag ccaccatcaa aagtgaaagt ggcttctaag gtactgtctt ctggacctca 1800
gagcccactc tccttggcac cctgacaccg ggaggcctca tggccaacaa tggacacctc 1860
cagagcctcc aggggaccac acagtagact atccctactc taagcagaga caactgccac 1920
ccagcctggg ccttcccaga ccagcatttg cacaatatca ccaggcttct tctgcctccc 1980
ttggtaaccc aaggaatgat ggaatcaaca caacatagta tgtttgcttt ccttacccaa 2040
ttgtaccttc tagaaaatca gtgttcacag agactgcctc caccacaggc atcctgcaaa 2100
tgcagactcc agaatcccca gcatcagcgg gaaccaccat cacatcttta ttcctcagcc 2160
cagacactcg aggcactcaa cagaatcagc catccacgtc taggtatcag agaggaccac 2220
aaatacaaca ttctccatct gctttcagag ttattatttt aataaaggaa gatctgggat 2280
gggctggtgg gccattccag cttgccgaaa tcaaagccat ctgaagcctg tctctggtga 2340
acaaacttcc tctctggcct ctcaggaatc agggtggcat ggctcacaac agcagggcct 2400
tcttcttttt gacgtgcaga atctcagtgg catctgggtt cacctcccca ctctgatgat 2460
ctccagcctc cactgcttct gccccccgct gctgaaatca aacatacccc aagttaaaat 2520
gaagctcccc cacccccact cccggccccg gttcccacag gacacgctaa gaagcacagg 2580
gagcatttaa caggctcacc ctccctttcc ttttcccctc ttctaccctc cccaagaaaa 2640
agggccttca aggcaggaat gagaaagcaa agccaatctc tcatttagac ctggcttctt 2700
tcttctgaac aaagtagggt tcaaaatgca gactgtcata tccagcgagt ccctgaccct 2760
ttctgcgaat gtaacgagca agcagtcagc acagcctggg ctgccctggc ccgggattga 2820
tgtagccccg gtaggtttgc ctctgcagaa ctaatggctg tgacttcaga gaaagccctg 2880
caggaagttt aacctgcgtg tcatctgcct ggtcatctca gacccatgaa attaggcgcc 2940
ttgtttgagc tgcgtttcac acttctttag agctagctga cctttggcca aaaataaact 3000
ttgaaaag 3008
<210>66
<211>560
<212>PRT
<213>Homo sapiens
<220>
<223>hyaluronan-binding serine protease(HGF activator-like protein)
<400>66
Met Phe Ala Arg Met Ser Asp Leu His Val Leu Leu Leu Met Ala Leu
1 5 10 15
Val Gly Lys Thr Ala Cys Gly Phe Ser Leu Met Ser Leu Leu Glu Ser
20 25 30
Leu Asp Pro Asp Trp Thr Pro Asp Gln Tyr Asp Tyr Ser Tyr Glu Asp
35 40 45
Tyr Asn Gln Glu Glu Asn Thr Ser Ser Thr Leu Thr His Ala Glu Asn
50 55 60
Pro Asp Trp Tyr Tyr Thr Glu Asp Gln Ala Asp Pro Cys Gln Pro Asn
65 70 75 80
Pro Cys Glu His Gly Gly Asp Cys Leu Val His Gly Ser Thr Phe Thr
85 90 95
Cys Ser Cys Leu Ala Pro Phe Ser Gly Asn Lys Cys Gln Lys Val Gln
100 105 110
Asn Thr Cys Lys Asp Asn Pro Cys Gly Arg Gly Gln Cys Leu Ile Thr
115 120 125
Gln Ser Pro Pro Tyr Tyr Arg Cys Val Cys Lys His Pro Tyr Thr Gly
130 135 140
Pro Ser Cys Ser Gln Val Val Pro Val Cys Arg Pro Asn Pro Cys Gln
145 150 155 160
Asn Gly Ala Thr Cys Ser Arg His Lys Arg Arg Ser Lys Phe Thr Cys
165 170 175
Ala Cys Pro Asp Gln Phe Lys Gly Lys Phe Cys Glu Ile Gly Ser Asp
180 185 190
Asp Cys Tyr Val Gly Asp Gly Tyr Ser Tyr Arg Gly Lys Met Asn Arg
195 200 205
Thr Val Asn Gln His Ala Cys Leu Tyr Trp Asn Ser His Leu Leu Leu
210 215 220
Gln Glu Asn Tyr Asn Met Phe Met Glu Asp Ala Glu Thr His Gly Ile
225 230 235 240
Gly Glu His Asn Phe Cys Arg Asn Pro Asp Ala Asp Glu Lys Pro Trp
245 250 255
Cys Phe Ile Lys Val Thr Asn Asp Lys Val Lys Trp Glu Tyr Cys Asp
260 265 270
Val Ser Ala Cys Ser Ala Gln Asp Val Ala Tyr Pro Glu Glu Ser Pro
275 280 285
Thr Glu Pro Ser Thr Lys Leu Pro Gly Phe Asp Ser Cys Gly Lys Thr
290 295 300
Glu Ile Ala Glu Arg Lys Ile Lys Arg Ile Tyr Gly Gly Phe Lys Ser
305 310 315 320
Thr Ala Gly Lys His Pro Trp Gln Ala Ser Leu Gln Ser Ser Leu Pro
325 330 335
Leu Thr Ile Ser Met Pro Gln Gly His Phe Cys Gly Gly Ala Leu Ile
340 345 350
His Pro Cys Trp Val Leu Thr Ala Ala His Cys Thr Asp Ile Lys Thr
355 360 365
Arg His Leu Lys Val Val Leu Gly Asp Gln Asp Leu Lys Lys Glu Glu
370 375 380
Phe His Glu Gln Ser Phe Arg Val Glu Lys Ile Phe Lys Tyr Ser His
385 390 395 400
Tyr Asn Glu Arg Asp Glu Ile Pro His Asn Asp Ile Ala Leu Leu Lys
405 410 415
Leu Lys Pro Val Asp Gly His Cys Ala Leu Glu Ser Lys Tyr Val Lys
420 425 430
Thr Val Cys Leu Pro Asp Gly Ser Phe Pro Ser Gly Ser Glu Cys His
435 440 445
Ile Ser Gly Trp Gly Val Thr Glu Thr Gly Lys Gly Ser Arg Gln Leu
450 455 460
Leu Asp Ala Lys Val Lys Leu Ile Ala Asn Thr Leu Cys Asn Ser Arg
465 470 475 480
Gln Leu Tyr Asp His Met Ile Asp Asp Ser Met Ile Cys Ala Gly Asn
485 490 495
Leu Gln Lys Pro Gly Gln Asp Thr Cys Gln Gly Asp Ser Gly Gly Pro
500 505 510
Leu Thr Cys Glu Lys Asp Gly Thr Tyr Tyr Val Tyr Gly Ile Val Ser
515 520 525
Trp Gly Leu Glu Cys Gly Lys Arg Pro Gly Val Tyr Thr Gln Val Thr
530 535 540
Lys Phe Leu Asn Trp Ile Lys Ala Thr Ile Lys Ser Glu Ser Gly Phe
545 550 555 560
<210>67
<211>2124
<212>DNA
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 4 transcript variant 1
<400>67
aatcaagctg cccaaagtcc cccaatcact cctggaatac acagagagag gcagcagctt 60
gctcagcgga caaggatgct gggcgtgagg gaccaaggcc tgccctgcac tcgggcctcc 120
tccagccagt gctgaccagg gacttctgac ctgctggcca gccaggacct gtgtggggag 180
gccctcctgc tgccttgggg tgacaatctc agctccaggc tacagggaga ccgggaggat 240
cacagagcca gcatgttaca ggatcctgac agtgatcaac ctctgaacag cctcgatgtc 300
aaacccctgc gcaaaccccg tatccccatg gagaccttca gaaaggtggg gatccccatc 360
atcatagcac tactgagcct ggcgagtatc atcattgtgg ttgtcctcat caaggtgatt 420
ctggataaat actacttcct ctgcgggcag cctctccact tcatcccgag gaagcagctg 480
tgtgacggag agctggactg tcccttgggg gaggacgagg agcactgtgt caagagcttc 540
cccgaagggc ctgcagtggc agtccgcctc tccaaggacc gatccacact gcaggtgctg 600
gactcggcca cagggaactg gttctctgcc tgtttcgaca acttcacaga agctctcgct 660
gagacagcct gtaggcagat gggctacagc agcaaaccca ctttcagagc tgtggagatt 720
ggcccagacc aggatctgga tgttgttgaa atcacagaaa acagccagga gcttcgcatg 780
cggaactcaa gtgggccctg tctctcaggc tccctggtct ccctgcactg tcttgcctgt 840
gggaagagcc tgaagacccc ccgtgtggtg ggtggggagg aggcctctgt ggattcttgg 900
ccttggcagg tcagcatcca gtacgacaaa cagcacgtct gtggagggag catcctggac 960
ccccactggg tcctcacggc agcccactgc ttcaggaaac ataccgatgt gttcaactgg 1020
aaggtgcggg caggctcaga caaactgggc agcttcccat ccctggctgt ggccaagatc 1080
atcatcattg aattcaaccc catgtacccc aaagacaatg acatcgccct catgaagctg 1140
cagttcccac tcactttctc aggcacagtc aggcccatct gtctgccctt ctttgatgag 1200
gagctcactc cagccacccc actctggatc attggatggg gctttacgaa gcagaatgga 1260
gggaagatgt ctgacatact gctgcaggcg tcagtccagg tcattgacag cacacggtgc 1320
aatgcagacg atgcgtacca gggggaagtc accgagaaga tgatgtgtgc aggcatcccg 1380
gaagggggtg tggacacctg ccagggtgac agtggtgggc ccctgatgta ccaatctgac 1440
cagtggcatg tggtgggcat cgttagctgg ggctatggct gcgggggccc gagcacccca 1500
ggagtataca ccaaggtctc agcctatctc aactggatct acaatgtctg gaaggctgag 1560
ctgtaatgct gctgcccctt tgcagtgctg ggagccgctt ccttcctgcc ctgcccacct 1620
ggggatcccc caaagtcaga cacagagcaa gagtcccctt gggtacaccc ctctgcccac 1680
agcctcagca tttcttggag cagcaaaggg cctcaattcc tgtaagagac cctcgcagcc 1740
cagaggcgcc cagaggaagt cagcagccct agctcggcca cacttggtgc tcccagcatc 1800
ccagggagag acacagccca ctgaacaagg tctcaggggt attgctaagc caagaaggaa 1860
ctttcccaca ctactgaatg gaagcaggct gtcttgtaaa agcccagatc actgtgggct 1920
ggagaggaga aggaaagggt ctgcgccagc cctgtccgtc ttcacccatc cccaagccta 1980
ctagagcaag aaaccagttg taatataaaa tgcactgccc tactgttggt atgactaccg 2040
ttacctactg ttgtcattgt tattacagct atggccacta ttattaaaga gctgtgtaac 2100
atctctggaa aaaaaaaaaa aaaa 2124
<210>68
<211>437
<212>PRT
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 4 isoform 1
<400>68
Met Leu Gln Asp Pro Asp Ser Asp Gln Pro Leu Asn Ser Leu Asp Val
1 5 10 15
Lys Pro Leu Arg Lys Pro Arg Ile Pro Met Glu Thr Phe Arg Lys Val
20 25 30
Gly Ile Pro Ile Ile Ile Ala Leu Leu Ser Leu Ala Ser Ile Ile Ile
35 40 45
Val Val Val Leu Ile Lys Val Ile Leu Asp Lys Tyr Tyr Phe Leu Cys
50 55 60
Gly Gln Pro Leu His Phe Ile Pro Arg Lys Gln Leu Cys Asp Gly Glu
65 70 75 80
Leu Asp Cys Pro Leu Gly Glu Asp Glu Glu His Cys ValLys Ser Phe
85 90 95
Pro Glu Gly Pro Ala Val Ala Val Arg Leu Ser Lys Asp Arg Ser Thr
100 105 110
Leu Gln Val Leu Asp Ser Ala Thr Gly Asn Trp Phe Ser Ala Cys Phe
115 120 125
Asp Asn Phe Thr Glu Ala Leu Ala Glu Thr Ala Cys Arg Gln Met Gly
130 135 140
Tyr Ser Ser Lys Pro Thr Phe Arg Ala ValGlu Ile Gly Pro Asp Gln
145 150 155 160
Asp Leu Asp Val Val Glu Ile Thr Glu Asn Ser Gln Glu Leu Arg Met
165 170 175
Arg Asn Ser Ser Gly Pro Cys Leu Ser Gly Ser Leu Val Ser Leu His
180 185 190
Cys Leu Ala Cys Gly Lys Ser Leu Lys Thr Pro Arg Val Val Gly Gly
195 200 205
Glu Glu Ala Ser Val Asp Ser Trp Pro Trp Gln Val Ser Ile Gln Tyr
210 215 220
Asp Lys Gln His Val Cys Gly Gly Ser Ile Leu Asp Pro His Trp Val
225 230 235 240
Leu Thr Ala Ala His Cys Phe Arg Lys His Thr Asp Val Phe Asn Trp
245 250 255
Lys Val Arg Ala Gly Ser Asp Lys Leu Gly Ser Phe Pro Ser Leu Ala
260 265 270
Val Ala Lys Ile Ile Ile Ile Glu Phe Asn Pro Met Tyr Pro Lys Asp
275 280 285
Asn Asp Ile Ala Leu Met Lys Leu Gln Phe Pro Leu Thr Phe Ser Gly
290 295 300
Thr Val Arg Pro Ile Cys Leu Pro Phe Phe Asp Glu Glu Leu Thr Pro
305 310 315 320
Ala Thr Pro Leu Trp Ile Ile Gly Trp Gly Phe Thr Lys Gln Asn Gly
325 330 335
Gly Lys Met Ser Asp Ile Leu Leu Gln Ala Ser Val Gln Val Ile Asp
340 345 350
Ser Thr Arg Cys Asn Ala Asp Asp Ala Tyr Gln Gly Glu Val Thr Glu
355 360 365
Lys Met Met Cys Ala Gly Ile Pro Glu Gly Gly Val Asp Thr Cys Gln
370 375 380
Gly Asp Ser Gly Gly Pro Leu Met Tyr Gln Ser Asp Gln Trp His Val
385 390 395 400
Val Gly Ile Val Ser Trp Gly Tyr Gly Cys Gly Gly Pro Ser Thr Pro
405 410 415
Gly Val Tyr Thr Lys Val Ser Ala Tyr Leu Asn Trp Ile Tyr Asn Val
420 425 430
Trp Lys Ala Glu Leu
435
<210>69
<211>1983
<212>DNA
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 4 transcript variant 2
<400>69
aatcaagctg cccaaagtcc cccaatcact cctggaatac acagagagag gcagcagctt 60
gctcagcgga caaggatgct gggcgtgagg gaccaaggcc tgccctgcac tcgggcctcc 120
tccagccagt gctgaccagg gacttctgac ctgctggcca gccaggacct gtgtggggag 180
gccctcctgc tgccttgggg tgacaatctc agctccaggc tacagggaga ccgggaggat 240
cacagagcca gcatggatcc tgacagtgat caacctctga acagcctcga tgtcaaaccc 300
ctgcgcaaac cccgtatccc catggagacc ttcagaaagg tggggatccc catcatcata 360
gcactactga gcctggcgag tatcatcatt gtggttgtcc tcatcaaggt gattctggat 420
aaatactact tcctctgcgg gcagcctctc cacttcatcc cgaggaagca gctgtgtgac 480
ggagagctgg actgtccctt gggggaggac gaggagcact gtgtcaagag cttccccgaa 540
gggcctgcag tggcagtccg cctctccaag gaccgatcca cactgcaggt gctggactcg 600
gccacaggga actggttctc tgcctgtttc gacaacttca cagaagctct cgctgagaca 660
gcctgtaggc agatgggcta cagcagcaaa cccactttca gagctgtgga gattggccca 720
gaccaggatc tggatgttgt tgaaatcaca gaaaacagcc aggagcttcg catgcggaac 780
tcaagtgggc cctgtctctc aggctccctg gtctccctgc actgtcttgc ctgtgggaag 840
agcctgaaga ccccccgtgt ggtgggtggg gaggaggcct ctgtggattc ttggccttgg 900
caggtcagca tccagtacga caaacagcac gtctgtggag ggagcatcct ggacccccac 960
tgggtcctca cggcagccca ctgcttcagg aaacataccg atgtgttcaa ctggaaggtg 1020
cgggcaggct cagacaaact gggcagcttc ccatccctgg ctgtggccaa gatcatcatc 1080
attgaattca accccatgta ccccaaagac aatgacatcg ccctcatgaa gctgcagttc 1140
ccactcactt tctcaggcac agtcaggccc atctgtctgc ccttctttga tgaggagctc 1200
actccagcca ccccactctg gatcattgga tggggcttta cgaagcagaa tggagggtga 1260
cagtggtggg cccctgatgt accaatctga ccagtggcat gtggtgggca tcgttagctg 1320
gggctatggc tgcgggggcc cgagcacccc aggagtatac accaaggtct cagcctatct 1380
caactggatc tacaatgtct ggaaggctga gctgtaatgc tgctgcccct ttgcagtgct 1440
gggagccgct tccttcctgc cctgcccacc tggggatccc ccaaagtcag acacagagca 1500
agagtcccct tgggtacacc cctctgccca cagcctcagc atttcttgga gcagcaaagg 1560
gcctcaattc ctgtaagaga ccctcgcagc ccagaggcgc ccagaggaag tcagcagccc 1620
tagctcggcc acacttggtg ctcccagcat cccagggaga gacacagccc actgaacaag 1680
gtctcagggg tattgctaag ccaagaagga actttcccac actactgaat ggaagcaggc 1740
tgtcttgtaa aagcccagat cactgtgggc tggagaggag aaggaaaggg tctgcgccag 1800
ccctgtccgt cttcacccat ccccaagcct actagagcaa gaaaccagtt gtaatataaa 1860
atgcactgcc ctactgttgg tatgactacc gttacctact gttgtcattg ttattacagc 1920
tatggccact attattaaag agctgtgtaa catctctgga aaaaaaaaaa aaaaaaaaaa 1980
aaa 1983
<210>70
<211>335
<212>PRT
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 4 isoform 2
<400>70
Met Asp Pro Asp Ser Asp Gln Pro Leu Asn Ser Leu Asp Val Lys Pro
1 5 10 15
Leu Arg Lys Pro Arg Ile Pro Met Glu Thr Phe Arg Lys Val Gly Ile
20 25 30
Pro Ile Ile Ile Ala Leu Leu Ser Leu Ala Ser Ile Ile Ile Val Val
35 40 45
Val Leu Ile Lys Val Ile Leu Asp Lys Tyr Tyr Phe Leu Cys Gly Gln
50 55 60
Pro Leu His Phe Ile Pro Arg Lys Gln Leu Cys Asp Gly Glu Leu Asp
65 70 75 80
Cys Pro Leu Gly Glu Asp Glu Glu His Cys Val Lys Ser Phe Pro Glu
85 90 95
Gly Pro Ala Val Ala Val Arg Leu Ser Lys Asp Arg Ser Thr Leu Gln
100 105 110
Val Leu Asp Ser Ala Thr Gly Asn Trp Phe Ser Ala Cys Phe Asp Asn
115 120 125
Phe Thr Glu Ala Leu Ala Glu Thr Ala Cys Arg Gln Met Gly Tyr Ser
130 135 140
Ser Lys Pro Thr Phe Arg Ala Val Glu Ile Gly Pro Asp Gln Asp Leu
145 150 155 160
Asp Val ValGlu Ile Thr Glu Asn Ser Gln Glu Leu Arg Met Arg Asn
165 170 175
Ser Ser Gly Pro Cys Leu Ser Gly Ser Leu Val Ser Leu His Cys Leu
180 185 190
Ala Cys Gly Lys Ser Leu Lys Thr Pro Arg Val Val Gly Gly Glu Glu
195 200 205
Ala Ser Val Asp Ser Trp Pro Trp Gln Val Ser Ile Gln Tyr Asp Lys
210 215 220
Gln His Val Cys Gly Gly Ser Ile Leu Asp Pro His Trp Val Leu Thr
225 230 235 240
Ala Ala His Cys Phe Arg Lys His Thr Asp Val Phe Asn Trp Lys Val
245 250 255
Arg Ala Gly Ser Asp Lys Leu Gly Ser Phe Pro Ser Leu Ala Val Ala
260 265 270
Lys Ile Ile Ile Ile Glu Phe Asn Pro Met Tyr Pro Lys Asp Asn Asp
275 280 285
Ile Ala Leu Met Lys Leu Gln Phe Pro Leu Thr Phe Ser Gly Thr Val
290 295 300
Arg Pro Ile Cys Leu Pro Phe Phe Asp Glu Glu Leu Thr Pro Ala Thr
305 310 315 320
Pro Leu Trp Ile Ile Gly Trp Gly Phe Thr Lys Gln Asn Gly Gly
325 330 335
<210>71
<211>855
<212>DNA
<213>Homo sapiens
<220>
<223>tryptase delta 1
<400>71
aggcccacga tgctcctcct tgctccccag atgctgagcc tgctgctgct ggcgctgccc 60
gtcctggcga gcccggccta cgtggcccct gccccaggcc aggccctgca gcaaacgggc 120
attgttgggg ggcaggaggc ccccaggagc aagtggccct ggcaggtgag cctgagagtc 180
cgcggcccat actggatgca cttctgcggg ggctccctca tccaccccca gtgggtgcta 240
accgcggcgc actgcgtgga accggacatc aaggatctgg ccgccctcag ggtgcaactg 300
cgggagcagc acctctacta ccaggaccag ctgctgccgg tcagcaggat catcgtgcac 360
ccacagttct acatcatcca gaccggggcg gacatcgccc tgctggagct ggaggagccc 420
gtgaacatct ccagccacat ccacacggtc acgctgcccc ctgcctcgga gaccttcccc 480
ccggggatgc cgtgctgggt cactggctgg ggcgacgtgg acaataatgt gcacctgccg 540
ccgccatacc cgctgaagga ggtggaagtc cccgtagtgg aaaaccacct ttgcaacgcg 600
gaatatcaca ccggcctcca tacgggccac agctttcaaa tcgtccgcga tgacatgctg 660
tgtgcgggga gcgaaaatca cgactcctgc cagggtgact ctggagggcc cctggtctgc 720
aaggtgaatg gcacctaact gcaggcgggc gtggtcagct gggaggagag ctgtgcccag 780
cccaaccggc ctggcatcta cacccgtgtc acctactact tggactggat ccaccactat 840
gtccccaaga agccc 855
<210>72
<211>242
<212>PRT
<213>Homo sapiens
<220>
<223>tryptase delta 1
<400>72
Met Leu Leu Leu Ala Pro Gln Met Leu Ser Leu Leu Leu Leu Ala Leu
1 5 10 15
Pro Val Leu Ala Ser Pro Ala Tyr Val Ala Pro Ala Pro Gly Gln Ala
20 25 30
Leu Gln Gln Thr Gly Ile Val Gly Gly Gln Glu Ala Pro Arg Ser Lys
35 40 45
Trp Pro Trp Gln Val Ser Leu Arg Val Arg Gly Pro Tyr Trp Met His
50 55 60
Phe Cys Gly Gly Ser Leu Ile His Pro Gln Trp Val Leu Thr Ala Ala
65 70 75 80
His Cys Val Glu Pro Asp Ile Lys Asp Leu Ala Ala Leu Arg Val Gln
85 90 95
Leu Arg Glu Gln His Leu Tyr Tyr Gln Asp Gln Leu Leu Pro Val Ser
100 105 110
Arg Ile Ile Val His Pro Gln Phe Tyr Ile Ile Gln Thr Gly Ala Asp
115 120 125
Ile Ala Leu Leu Glu Leu Glu Glu Pro Val Asn Ile Ser Ser His Ile
130 135 140
His Thr Val Thr Leu Pro Pro Ala Ser Glu Thr Phe Pro Pro Gly Met
145 150 155 160
Pro Cys Trp Val Thr Gly Trp Gly Asp Val Asp Asn Asn Val His Leu
165 170 175
Pro Pro Pro Tyr Pro Leu Lys Glu Val Glu Val Pro Val Val Glu Asn
180 185 190
His Leu Cys Asn Ala Glu Tyr His Thr Gly Leu His Thr Gly His Ser
195 200 205
Phe Gln Ile Val Arg Asp Asp Met Leu Cys Ala Gly Ser Glu Asn His
210 215 220
Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Lys Val Asn
225 230 235 240
Gly Thr
<210>73
<211>1462
<212>DNA
<213>Homo sapiens
<220>
<223>Marapsin
<400>73
ctgtctcaaa aaattaaaaa aaataataat aataataatg aatgaagctg gacggacttc 60
gcgtgcaccg cggtcagctc ggggtctgct ggggggtctg ggtcagctca gggtccagga 120
accgaggcca acggcacccc gtgctgcgct ggggtgaggg gtctgccctg gggtctcggg 180
gttcagggct aggtcacgga ggagtcggct ctgggcgctt ccttcctgag gagaggagct 240
gggcaggccg ggccgacggg ttgggccgca tagccgggcc tgtgctcatc tccagcataa 300
aactccactt catggagcct gcacctcgct cgtgctccaa cgcttctgcc accgccgacc 360
acggccctgc gccccagcca ggcctgagga catgaggcgg ccggcggcgg tgccgctcct 420
gctgctgctg tgttttgggt ctcagagggc caaggcagca acagcctgtg gtcgccccag 480
gatgctgaac cgaatggtgg gcgggcagga cacgcaggag ggcgagtggc cctggcaagt 540
cagcatccag cgcaacggaa gccacttctg cgggggcagc ctcatcgcgg agcagtgggt 600
cctgacggct gcgcactgct tccgcaacac ctctgagacg tccctgtacc aggtcctgct 660
gggggcaagg cagctagtgc agccgggacc acacgctatg tatgcccggg tgaggcaggt 720
ggagagcaac cccctgtacc agggcacggc ctccagcgct gacgtggccc tggtggagct 780
ggaggcacca gtgcccttca ccaattacat cctccccgtg tgcctgcctg acccctcggt 840
gatctttgag acgggcatga actgctgggt cactggctgg ggcagcccca gtgaggaaga 900
cctcctgccc gaaccgcgga tcctgcagaa actcgctgtg cccatcatcg acacacccaa 960
gtgcaacctg ctctacagca aagacaccga gtttggctac caacccaaaa ccatcaagaa 1020
tgacatgctg tgcgccggct tcgaggaggg caagaaggat gcctgcaagg gcgactcggg 1080
cggccccctg gtgtgcctcg tgggtcagtc gtggctgcag gcgggggtga tcagctgggg 1140
tgagggctgt gcccgccaga accgcccagg tgtctacatc cgtgtcaccg cccaccacaa 1200
ctggatccat cggatcatcc ccaaactgca gttccagcca gcgaggttgg gcggccagaa 1260
gtgagacccc cggggccagg agccccttga gcagagctct gcacccagcc tgcccgccca 1320
caccatcctg ctggtcctcc cagcgctgct gttgcacctg tgagccccac cagactcatt 1380
tgtaaatagc gctccttcct cccctctcaa atacccttat tttatttatg tttctcccaa 1440
taaaaaccca gcctgtgtgc ca 1462
<210>74
<211>290
<212>PRT
<213>Homo sapiens
<220>
<223>Marapsin
<400>74
Met Arg Arg Pro Ala Ala Val Pro Leu Leu Leu Leu Leu Cys Phe Gly
1 5 10 15
Ser Gln Arg Ala Lys Ala Ala Thr Ala Cys Gly Arg Pro Arg Met Leu
20 25 30
Asn Arg Met Val Gly Gly Gln Asp Thr Gln Glu Gly Glu Trp Pro Trp
35 40 45
Gln Val Ser Ile Gln Arg Asn Gly Ser His Phe Cys Gly Gly Ser Leu
50 55 60
Ile Ala Glu Gln Trp Val Leu Thr Ala Ala His Cys Phe Arg Asn Thr
65 70 75 80
Ser Glu Thr Ser Leu Tyr Gln Val Leu Leu Gly Ala Arg Gln Leu Val
85 90 95
Gln Pro Gly Pro His Ala Met Tyr Ala Arg Val Arg Gln Val Glu Ser
100 105 110
Asn Pro Leu Tyr Gln Gly Thr Ala Ser Ser Ala Asp Val Ala Leu Val
115 120 125
Glu Leu Glu Ala Pro Val Pro Phe Thr Asn Tyr Ile Leu Pro Val Cys
130 135 140
Leu Pro Asp Pro Ser Val Ile Phe Glu Thr Gly Met Asn Cys Trp Val
145 150 155 160
Thr Gly Trp Gly Ser Pro Ser Glu Glu Asp Leu Leu Pro Glu Pro Arg
165 170 175
Ile Leu Gln Lys Leu Ala Val Pro Ile Ile Asp Thr Pro Lys Cys Asn
180 185 190
Leu Leu Tyr Ser Lys Asp Thr Glu Phe Gly Tyr Gln Pro Lys Thr Ile
195 200 205
Lys Asn Asp Met Leu Cys Ala Gly Phe Glu Glu Gly Lys Lys Asp Ala
210 215 220
Cys Lys Gly Asp Ser Gly Gly Pro Leu Val Cys Leu Val Gly Gln Ser
225 230 235 240
Trp Leu Gln Ala Gly Val Ile Ser Trp Gly Glu Gly Cys Ala Arg Gln
245 250 255
Asn Arg Pro Gly Val Tyr Ile Arg Val Thr Ala His His Asn Trp Ile
260 265 270
His Arg Ile Ile Pro Lys Leu Gln Phe Gln Pro Ala Arg Leu Gly Gly
275 280 285
Gln Lys
290
<210>75
<211>1755
<212>DNA
<213>Homo sapiens
<220>
<223>Tryptase homologue 2
<400>75
ccaggacttc cgtctgagat gggatttctg gacagaacaa tggagcgcag aagctccagc 60
aacgtgtgct ctgcagggag cccagaccag agtccaagcc ctaggcagtg ccacccttac 120
ccagcccagc cttgaagaca gaatgagagg ggtttcctgt ctccaggtcc tgctccttct 180
ggtgctggga gctgctggga ctcagggaag gaagtctgca gcctgcgggc agccccgcat 240
gtccagtcgg atcgttgggg gccgggatgg ccgggacgga gagtggccgt ggcaggcgag 300
catccagcat cgtggggcac acgtgtgcgg ggggtcgctc atcgcccccc agtgggtgct 360
gacagcggcg cactgcttcc ccaggagggc actgccagct gagtaccgcg tgcgcctggg 420
ggcgctgcgt ctgggctcca cctcgccccg cacgctctcg gtgcccgtgc gacgggtgct 480
gctgcccccg gactactccg aggacggggc ccgcggcgac ctggcactgc tgcagctgcg 540
tcgcccggtg cccctgagcg ctcgcgtcca acccgtctgc ctgcccgtgc ccggcgcccg 600
cccgccgccc ggcacaccat gccgggtcac cggctggggc agcctccgcc caggagtgcc 660
cctcccagag tggcgaccgc tacaaggagt aagggtgccg ctgctggact cgcgcacctg 720
cgacggcctc taccacgtgg gcgcggacgt gccccaggct gagcgcattg tgctgcctgg 780
gagtctgtgt gccggctacc cccagggcca caaggacgcc tgccagggtg attctggggg 840
acctctgacc tgcctgcagt ctgggagctg ggtcctggtg ggcgtggtga gctggggcaa 900
gggttgtgcc ctgcccaacc gtccaggggt ctacaccagt gtggccacat atagcccctg 960
gattcaggct cgcgtcagct tctaatgcta gccggtgagg ctgacctgga gccagctgct 1020
ggggtccctc agcctcctgg ttcatccagg cacctgccta taccccacat cccttctgcc 1080
tcgaggccaa gatgcctaaa aaagctaaag gccaccccac cccccaccca ccacctcctg 1140
cctcctctcc tctttgggga tcaccagctc tgactccacc aaccctcatc caggaatctg 1200
ccatgagtcc cagggagtca cactccccac tcccttcctg gcttgtattt acttttcttg 1260
gccctggcca gggctgggcg caaggcacgc agtgatgggc aaaccaattg ctgcccatct 1320
ggcctgtgtg cccatctttt tctggagaaa gtcagattca cagcatgaca gagatttgac 1380
accagggaga tcctccatag ctggctttga ggacacgggg accacagcca tgagcggcct 1440
ctaagagctg agagacagcc ggcagggaat cggaaccctc agacccacag ccgcaaggca 1500
ctggattctg gcagcaccct gaaggagctg ggaagtaagt tcttccccag cctccagata 1560
agagccccgc cggccaatcc cttcatttca acctaaagag accctaagca gagaacctag 1620
ctgagccact cctgacctac aaagttgtga cttaataaat gtgtgctttg aacccaaaaa 1680
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1740
aaaaaaaaaa aaaaa 1755
<210>76
<211>284
<212>PRT
<213>Homo sapiens
<220>
<223>Tryptase homologue 2
<400>76
Met Arg Gly Val Ser Cys Leu Gln Val Leu Leu Leu Leu Val Leu Gly
1 5 10 15
Ala Ala Gly Thr Gln Gly Arg Lys Ser Ala Ala Cys Gly Gln Pro Arg
20 25 30
Met Ser Ser Arg Ile Val Gly Gly Arg Asp Gly Arg Asp Gly Glu Trp
35 40 45
Pro Trp Gln Ala Ser Ile Gln His Pro Gly Ala His Val Cys Gly Gly
50 55 60
Ser Leu Ile Ala Pro Gln Trp Val Leu Thr Ala Ala His Cys Phe Pro
65 70 75 80
Arg Arg Ala Leu Pro Ala Glu Tyr Arg Val Arg Leu Gly Ala Leu Arg
85 90 95
Leu Gly Ser Thr Ser Pro Arg Thr Leu Ser Val Pro Val Arg Arg Val
100 105 110
Leu Leu Pro Pro Asp Tyr Ser Glu Asp Gly Ala Arg Gly Asp Leu Ala
115 120 125
Leu Leu Gln Leu Arg Arg Pro Val Pro Leu Ser Ala Arg Val Gln Pro
130 135 140
Val Cys Leu Pro Val Pro Gly Ala Arg Pro Pro Pro Gly Thr Pro Cys
145 150 155 160
Arg Val Thr Gly Trp Gly Ser Leu Arg Pro Gly Val Pro Leu Pro Glu
165 170 175
Trp Arg Pro Leu Gln Gly Val Arg Val Pro Leu Leu Asp Ser Arg Thr
180 185 190
Cys Asp Gly Leu Tyr His Val Gly Ala Asp Val Pro Gln Ala Glu Arg
195 200 205
Ile Val Leu Pro Gly Ser Leu Cys Ala Gly Tyr Pro Gln Gly His Lys
210 215 220
Asp Ala Cys Gln Gly Asp Ser Gly Gly Pro Leu Thr Cys Leu Gln Ser
225 230 235 240
Gly Ser Trp Val Leu ValGly Val Val Ser Trp Gly Lys Gly Cys Ala
245 250 255
Leu Pro Asn Arg Pro Gly Val Tyr Thr Ser ValAla Thr Tyr Ser Pro
260 265 270
Trp Ile Gln Ala Arg Val Thr Ser Asn Ala Ser Arg
275 280
<210>77
<211>321
<212>DNA
<213>Homo sapiens
<220>
<223>Tryptase homologue 3
<400>77
gcgctggtgg cgggcggagt ggagtccgcg cgcgggcgct ggccatggca ggccagcctg 60
cgcctgagga gacgccaccg atgtggaggg agcctgctca gccgccgctg ggtgctctcg 120
gctgcgcact gcttcgctcc ccagggtgac tcaggtggac ccttggtctg tgacaaggat 180
ggactgtggt atcaggttgg aatcgtgagc tggggaatgg actgcggtca acccaatcgg 240
cctggtgtct acaccaacat cagtgtgtac ttccactgga tccggagggt gatgtcccac 300
agtacaccca ggccaaaccc c 321
<210>78
<211>107
<212>PRT
<213>Homo sapiens
<220>
<223>Tryptase homologue 3
<400>78
Ala Leu Val Ala Gly Gly Val Glu Ser Ala Arg Gly Arg Trp Pro Trp
1 5 10 15
Gln Ala Ser Leu Arg Leu Arg Arg Arg His Arg Cys Gly Gly Ser Leu
20 25 30
Leu Ser Arg Arg Trp Val Leu Ser Ala Ala His Cys Phe Ala Pro Gln
35 40 45
Gly Asp Ser Gly Gly Pro Leu Val Cys Asp Lys Asp Gly Leu Trp Tyr
50 55 60
Gln Val Gly Ile Val Ser Trp Gly Met Asp Cys Gly Gln Pro Asn Arg
65 70 75 80
Pro Gly Val Tyr Thr Asn Ile Ser Val Tyr Phe His Trp Ile Arg Arg
85 90 95
Val Met Ser His Ser Thr Pro Arg Pro Asn Pro
100 105
<210>79
<211>2468
<212>DNA
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 3 transcript variant A
<400>79
accgggcacc ggacggctcg ggtactttcg ttcttaatta ggtcatgccc gtgtgagcca 60
ggaaagggct gtgtttatgg gaagccagta acactgtggc ctactatctc ttccgtggtg 120
ccatctacat ttttgggact cgggaattat gaggtagagg tggaggcgga gccggatgtc 180
agaggtcctg aaatagtcac catgggggaa aatgatccgc ctgctgttga agcccccttc 240
tcattccgat cgctttttgg ccttgatgat ttgaaaataa gtcctgttgc accagatgca 300
gatgctgttg ctgcacagat cctgtcactg ctgccattga agttttttcc aatcatcgtc 360
attgggatca ttgcattgat attagcactg gccattggtc tgggcatcca cttcgactgc 420
tcagggaagt acagatgtcg ctcatccttt aagtgtatcg agctgatagc tcgatgtgac 480
ggagtctcgg attgcaaaga cggggaggac gagtaccgct gtgtccgggt gggtggtcag 540
aatgccgtgc tccaggtgtt cacagctgct tcgtggaaga ccatgtgctc cgatgactgg 600
aagggtcact acgcaaatgt tgcctgtgcc caactgggtt tcccaagcta tgtgagttca 660
gataacctca gagtgagctc gctggagggg cagttccggg aggagtttgt gtccatcgat 720
cacctcttgc cagatgacaa ggtgactgca ttacaccact cagtatatgt gagggaggga 780
tgtgcctctg gccacgtggt taccttgcag tgcacagcct gtggtcatag aaggggctac 840
agctcacgca tcgtgggtgg aaacatgtcc ttgctctcgc agtggccctg gcaggccagc 900
cttcagttcc agggctacca cctgtgcggg ggctctgtca tcacgcccct gtggatcatc 960
actgctgcac actgtgttta tgacttgtac ctccccaagt catggaccat ccaggtgggt 1020
ctagtttccc tgttggacaa tccagcccca tcccacttgg tggagaagat tgtctaccac 1080
agcaagtaca agccaaagag gctgggcaat gacatcgccc ttatgaagct ggccgggcca 1140
ctcacgttca atgaaatgat ccagcctgtg tgcctgccca actctgaaga gaacttcccc 1200
gatggaaaag tgtgctggac gtcaggatgg ggggccacag aggatggagc aggtgacgcc 1260
tcccctgtcc tgaaccacgc ggccgtccct ttgatttcca acaagatctg caaccacagg 1320
gacgtgtacg gtggcatcat ctccccctcc atgctctgcg cgggctacct gacgggtggc 1380
gtggacagct gccaggggga cagcgggggg cccctggtgt gtcaagagag gaggctgtgg 1440
aagttagtgg gagcgaccag ctttggcatc ggctgcgcag aggtgaacaa gcctggggtg 1500
tacacccgtg tcacctcctt cctggactgg atccacgagc agatggagag agacctaaaa 1560
acctgaagag gaaggggaca agtagccacc tgagttcctg aggtgatgaa gacagcccga 1620
tcctcccctg gactcccgtg taggaacctg cacacgagca gacacccttg gagctctgag 1680
ttccggcacc agtagcaggc ccgaaagagg cacccttcca tctgattcca gcacaacctt 1740
caagctgctt tttgtttttt gtttttttga ggtggagtct cgctctgttg cccaggctgg 1800
agtgcagtgg cgaaatccct gctcactgca gcctccgctt ccctggttca agcgattctc 1860
ttgcctcagc ttccccagta gctgggacca caggtgcccg ccaccacacc caactaattt 1920
ttgtattttt agtagagaca gggtttcacc atgttggcca ggctgctctc aaacccctga 1980
cctcaaatga tgtgcctgct tcagcctccc acagtgctgg gattacaggc atgggccacc 2040
acgcctagcc tcacgctcct ttctgatctt cactaagaac aaaagaagca gcaacttgca 2100
agggcggcct ttcccactgg tccatctggt tttctctcca ggggtcttgc aaaattcctg 2160
acgagataag cagttatgtg acctcacgtg caaagccacc aacagccact cagaaaagac 2220
gcaccagccc agaagtgcag aactgcagtc actgcacgtt ttcatctcta gggaccagaa 2280
ccaaacccac cctttctact tccaagactt attttcacat gtggggaggt taatctagga 2340
atgactcgtt taaggcctat tttcatgatt tctttgtagc atttggtgct tgacgtatta 2400
ttgtcctttg attccaaata atatgtttcc ttccctcaaa aaaaaaaaaa aaaaaaaaaa 2460
aaaaaaaa 2468
<210>80
<211>454
<212>PRT
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 3 isoform 1
<400>80
Met Gly Glu Asn Asp Pro Pro Ala Val Glu Ala Pro Phe Ser Phe Arg
1 5 10 15
Ser Leu Phe Gly Leu Asp Asp Leu Lys Ile Ser Pro Val Ala Pro Asp
20 25 30
Ala Asp Ala Val Ala Ala Gln Ile Leu Ser Leu Leu Pro Leu Lys Phe
35 40 45
Phe Pro Ile Ile Val Ile Gly Ile Ile Ala Leu Ile Leu Ala Leu Ala
50 55 60
Ile Gly Leu Gly Ile His Phe Asp Cys Ser Gly Lys Tyr Arg Cys Arg
65 70 75 80
Ser Ser Phe Lys Cys Ile Glu Leu Ile Ala Arg Cys Asp Gly Val Ser
85 90 95
Asp Cys Lys Asp Gly Glu Asp Glu Tyr Arg Cys Val Arg Val Gly Gly
100 105 110
Gln Asn Ala Val Leu Gln Val Phe Thr Ala Ala Ser Trp Lys Thr Met
115 120 125
Cys Ser Asp Asp Trp Lys Gly His Tyr Ala Asn Val Ala Cys Ala Gln
130 135 140
Leu Gly Phe Pro Ser Tyr Val Ser Ser Asp Asn Leu Arg Val Ser Ser
145 150 155 160
Leu Glu Gly Gln Phe Arg Glu Glu Phe Val Ser Ile Asp His Leu Leu
165 170 175
Pro Asp Asp Lys Val Thr Ala Leu His His Ser Val Tyr Val Arg Glu
180 185 190
Gly Cys Ala Ser Gly His Val Val Thr Leu Gln Cys Thr Ala Cys Gly
195 200 205
His Arg Arg Gly Tyr Ser Ser Arg Ile Val Gly Gly Asn Met Ser Leu
210 215 220
Leu Ser Gln Trp Pro Trp Gln Ala Ser Leu Gln Phe Gln Gly Tyr His
225 230 235 240
Leu Cys Gly Gly Ser Val Ile Thr Pro Leu Trp Ile Ile Thr Ala Ala
245 250 255
His Cys Val Tyr Asp Leu Tyr Leu Pro Lys Ser Trp Thr Ile Gln Val
260 265 270
Gly Leu Val Ser Leu Leu Asp Asn Pro Ala Pro Ser His Leu Val Glu
275 280 285
Lys Ile Val Tyr His Ser Lys Tyr Lys Pro Lys Arg Leu Gly Asn Asp
290 295 300
Ile Ala Leu Met Lys Leu Ala Gly Pro Leu Thr Phe Asn Glu Met Ile
305 310 315 320
Gln Pro Val Cys Leu Pro Asn Ser Glu Glu Asn Phe Pro Asp Gly Lys
325 330 335
Val Cys Trp Thr Ser Gly Trp Gly Ala Thr Glu Asp Gly Ala Gly Asp
340 345 350
Ala Ser Pro Val Leu Asn His Ala Ala Val Pro Leu Ile Ser Asn Lys
355 360 365
Ile Cys Asn His Arg Asp Val Tyr Gly Gly Ile Ile Ser Pro Ser Met
370 375 380
Leu Cys Ala Gly Tyr Leu Thr Gly Gly Val Asp Ser Cys Gln Gly Asp
385 390 395 400
Ser Gly Gly Pro Leu Val Cys Gln Glu Arg Arg Leu Trp Lys Leu Val
405 410 415
Gly Ala Thr Ser Phe Gly Ile Gly Cys Ala Glu Val Asn Lys Pro Gly
420 425 430
Val Tyr Thr Arg Val Thr Ser Phe Leu Asp Trp Ile His Glu Gln Met
435 440 445
Glu Arg Asp Leu Lys Thr
450
<210>81
<211>2554
<212>DNA
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 3 transcript variant B
<400>81
gccacgttgg attagggccc accctaatga ctcattttaa ctagatgact ctgtaaagac 60
tctccaggcc atctccaaat aagatgccat tctgaggtac taagagttaa gacctcaacg 120
tgtctttctg ggggatacta ttcaacccat agcacatccc ctcatccccc atggaataac 180
cgggtcagag gtggagcagg taggactgaa ctagagaatg tgccccttgg tgggtggccc 240
tttccctaat taaggcacct ttctttgggc tgagacttct ctgcaaagag gggaggggga 300
aaggatagtt aaatttgtaa gtttcatggc actccaagtc aggttctggg gctttctgct 360
cttccttttt ctggttatca gatgcagatg ctgttgctgc acagatcctg tcactgctgc 420
cattgaagtt ttttccaatc atcgtcattg ggatcattgc attgatatta gcactggcca 480
ttggtctggg catccacttc gactgctcag ggaagtacag atgtcgctca tcctttaagt 540
gtatcgagct gatagctcga tgtgacggag tctcggattg caaagacggg gaggacgagt 600
accgctgtgt ccgggtgggt ggtcagaatg ccgtgctcca ggtgttcaca gctgcttcgt 660
ggaagaccat gtgctccgat gactggaagg gtcactacgc aaatgttgcc tgtgcccaac 720
tgggtttccc aagctatgtg agttcagata acctcagagt gagctcgctg gaggggcagt 780
tccgggagga gtttgtgtcc atcgatcacc tcttgccaga tgacaaggtg actgcattac 840
accactcagt atatgtgagg gagggatgtg cctctggcca cgtggttacc ttgcagtgca 900
cagcctgtgg tcatagaagg ggctacagct cacgcatcgt gggtggaaac atgtccttgc 960
tctcgcagtg gccctggcag gccagccttc agttccaggg ctaccacctg tgcgggggct 1020
ctgtcatcac gcccctgtgg atcatcactg ctgcacactg tgtttatgac ttgtacctcc 1080
ccaagtcatg gaccatccag gtgggtctag tttccctgtt ggacaatcca gccccatccc 1140
acttggtgga gaagattgtc taccacagca agtacaagcc aaagaggctg ggcaatgaca 1200
tcgcccttat gaagctggcc gggccactca cgttcaatga aatgatccag cctgtgtgcc 1260
tgcccaactc tgaagagaac ttccccgatg gaaaagtgtg ctggacgtca ggatgggggg 1320
ccacagagga tggagcaggt gacgcctccc ctgtcctgaa ccacgcggcc gtccctttga 1380
tttccaacaa gatctgcaac cacagggacg tgtacggtgg catcatctcc ccctccatgc 1440
tctgcgcggg ctacctgacg ggtggcgtgg acagctgcca gggggacagc ggggggcccc 1500
tggtgtgtca agagaggagg ctgtggaagt tagtgggagc gaccagcttt ggcatcggct 1560
gcgcagaggt gaacaagcct ggggtgtaca cccgtgtcac ctccttcctg gactggatcc 1620
acgagcagat ggagagagac ctaaaaacct gaagaggaag gggacaagta gccacctgag 1680
ttcctgaggt gatgaagaca gcccgatcct cccctggact cccgtgtagg aacctgcaca 1740
cgagcagaca cccttggagc tctgagttcc ggcaccagta gcaggcccga aagaggcacc 1800
cttccatctg attccagcac aaccttcaag ctgctttttg ttttttgttt ttttgaggtg 1860
gagtctcgct ctgttgccca ggctggagtg cagtggcgaa atccctgctc actgcagcct 1920
ccgcttccct ggttcaagcg attctcttgc ctcagcttcc ccagtagctg ggaccacagg 1980
tgcccgccac cacacccaac taatttttgt atttttagta gagacagggt ttcaccatgt 2040
tggccaggct gctctcaaac ccctgacctc aaatgatgtg cctgcttcag cctcccacag 2100
tgctgggatt acaggcatgg gccaccacgc ctagcctcac gctcctttct gatcttcact 2160
aagaacaaaa gaagcagcaa cttgcaaggg cggcctttcc cactggtcca tctggttttc 2220
tctccagggg tcttgcaaaa ttcctgacga gataagcagt tatgtgacct cacgtgcaaa 2280
gccaccaaca gccactcaga aaagacgcac cagcccagaa gtgcagaact gcagtcactg 2340
cacgttttca tctctaggga ccagaaccaa acccaccctt tctacttcca agacttattt 2400
tcacatgtgg ggaggttaat ctaggaatga ctcgtttaag gcctattttc atgatttctt 2460
tgtagcattt ggtgcttgac gtattattgt cctttgattc caaataatat gtttccttcc 2520
ctcaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 2554
<210>82
<211>327
<212>PRT
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 3 isoform 2
<400>82
Met Cys Ser Asp Asp Trp Lys Gly His Tyr Ala Asn Val Ala Cys Ala
1 5 10 15
Gln Leu Gly Phe Pro Ser Tyr Val Ser Ser Asp Asn Leu Arg Val Ser
20 25 30
Ser Leu Glu Gly Gln Phe Arg Glu Glu Phe Val Ser Ile Asp His Leu
35 40 45
Leu Pro Asp Asp Lys Val Thr Ala Leu His His Ser Val Tyr Val Arg
50 55 60
Glu Gly Cys Ala Ser Gly His Val Val Thr Leu Gln Cys Thr Ala Cys
65 70 75 80
Gly His Arg Arg Gly Tyr Ser Ser Arg Ile Val Gly Gly Asn Met Ser
85 90 95
Leu Leu Ser Gln Trp Pro Trp Gln Ala Ser Leu Gln Phe Gln Gly Tyr
100 105 110
His Leu Cys Gly Gly Ser Val Ile Thr Pro Leu Trp Ile Ile Thr Ala
115 120 125
Ala His Cys Val Tyr Asp Leu Tyr Leu Pro Lys Ser Trp Thr Ile Gln
130 135 140
Val Gly Leu Val Ser Leu Leu Asp Asn Pro Ala Pro Ser His Leu Val
145 150 155 160
Glu Lys Ile Val Tyr His Ser Lys Tyr Lys Pro Lys Arg Leu Gly Asn
165 170 175
Asp Ile Ala Leu Met Lys Leu Ala Gly Pro Leu Thr Phe Asn Glu Met
180 185 190
Ile Gln Pro Val Cys Leu Pro Asn Ser Glu Glu Asn Phe Pro Asp Gly
195 200 205
Lys Val Cys Trp Thr Ser Gly Trp Gly Ala Thr Glu Asp Gly Ala Gly
210 215 220
Asp Ala Ser Pro Val Leu Asn His Ala Ala Val Pro Leu Ile Ser Asn
225 230 235 240
Lys Ile Cys Asn His Arg Asp ValTyr Gly Gly Ile Ile Ser Pro Ser
245 250 255
Met Leu Cys Ala Gly Tyr Leu Thr Gly Gly Val Asp Ser Cys Gln Gly
260 265 270
Asp Ser Gly Gly Pro Leu Val Cys Gln Glu Arg Arg Leu Trp Lys Leu
275 280 285
Val Gly Ala Thr Ser Phe Gly Ile Gly Cys Ala Glu Val Asn Lys Pro
290 295 300
Gly Val Tyr Thr Arg Val Thr Ser Phe Leu Asp Trp Ile His Glu Gln
305 310 315 320
Met Glu Arg Asp Leu Lys Thr
325
<210>83
<211>2135
<212>DNA
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 3 transcript variant C
<400>83
gcagggggac agttgttagt gttgcactct gaaagagctg ttggttaatt cctgccttcc 60
tcccttttcc cagtccactt cgactgctca gggaagtaca gatgtcgctc atcctttaag 120
tgtatcgagc tgatagctcg atgtgacgga gtctcggatt gcaaagacgg ggaggacgag 180
taccgctgtg tccgggtggg tggtcagaat gccgtgctcc aggtgttcac agctgcttcg 240
tggaagacca tgtgctccga tgactggaag ggtcactacg caaatgttgc ctgtgcccaa 300
ctgggtttcc caagctatgt gagttcagat aacctcagag tgagctcgct ggaggggcag 360
ttccgggagg agtttgtgtc catcgatcac ctcttgccag atgacaaggt gactgcatta 420
caccactcag tatatgtgag ggagggatgt gcctctggcc acgtggttac cttgcagtgc 480
acagcctgtg gtcatagaag gggctacagc tcacgcatcg tgggtggaaa catgtccttg 540
ctctcgcagt ggccctggca ggccagcctt cagttccagg gctaccacct gtgcgggggc 600
tctgtcatca cgcccctgtg gatcatcact gctgcacact gtgtttatga cttgtacctc 660
cccaagtcat ggaccatcca ggtgggtcta gtttccctgt tggacaatcc agccccatcc 720
cacttggtgg agaagattgt ctaccacagc aagtacaagc caaagaggct gggcaatgac 780
atcgccctta tgaagctggc cgggccactc acgttcaatg aaatgatcca gcctgtgtgc 840
ctgcccaact ctgaagagaa cttccccgat ggaaaagtgt gctggacgtc aggatggggg 900
gccacagagg atggagcagg tgacgcctcc cctgtcctga accacgcggc cgtccctttg 960
atttccaaca agatctgcaa ccacagggac gtgtacggtg gcatcatctc cccctccatg 1020
ctctgcgcgg gctacctgac gggtggcgtg gacagctgcc agggggacag cggggggccc 1080
ctggtgtgtc aagagaggag gctgtggaag ttagtgggag cgaccagctt tggcatcggc 1140
tgcgcagagg tgaacaagcc tggggtgtac acccgtgtca cctccttcct ggactggatc 1200
cacgagcaga tggagagaga cctaaaaacc tgaagaggaa ggggacaagt agccacctga 1260
gttcctgagg tgatgaagac agcccgatcc tcccctggac tcccgtgtag gaacctgcac 1320
acgagcagac acccttggag ctctgagttc cggcaccagt agcaggcccg aaagaggcac 1380
ccttccatct gattccagca caaccttcaa gctgcttttt gttttttgtt tttttgaggt 1440
ggagtctcgc tctgttgccc aggctggagt gcagtggcga aatccctgct cactgcagcc 1500
tccgcttccc tggttcaagc gattctcttg cctcagcttc cccagtagct gggaccacag 1560
gtgcccgcca ccacacccaa ctaatttttg tatttttagt agagacaggg tttcaccatg 1620
ttggccaggc tgctctcaaa cccctgacct caaatgatgt gcctgcttca gcctcccaca 1680
gtgctgggat tacaggcatg ggccaccacg cctagcctca cgctcctttc tgatcttcac 1740
taagaacaaa agaagcagca acttgcaagg gcggcctttc ccactggtcc atctggtttt 1800
ctctccaggg gtcttgcaaa attcctgacg agataagcag ttatgtgacc tcacgtgcaa 1860
agccaccaac agccactcag aaaagacgca ccagcccaga agtgcagaac tgcagtcact 1920
gcacgttttc atctctaggg accagaacca aacccaccct ttctacttcc aagacttatt 1980
ttcacatgtg gggaggttaa tctaggaatg actcgtttaa ggcctatttt catgatttct 2040
ttgtagcatt tggtgcttga cgtattattg tcctttgatt ccaaataata tgtttccttc 2100
cctcaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaa 2135
<210>84
<211>327
<212>PRT
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 3 isoform 2
<400>84
Met Cys Ser Asp Asp Trp Lys Gly His Tyr Ala Asn Val Ala Cys Ala
1 5 10 15
Gln Leu Gly Phe Pro Ser Tyr Val Ser Ser Asp Asn Leu Arg Val Ser
20 25 30
Ser Leu Glu Gly Gln Phe Arg Glu Glu Phe Val Ser Ile Asp His Leu
35 40 45
Leu Pro Asp Asp Lys Val Thr Ala Leu His His Ser Val Tyr Val Arg
50 55 60
Glu Gly Cys Ala Ser Gly His Val Val Thr Leu Gln Cys Thr Ala Cys
65 70 75 80
Gly His Arg Arg Gly Tyr Ser Ser Arg Ile ValGly Gly Asn Met Ser
85 90 95
Leu Leu Ser Gln Trp Pro Trp Gln Ala Ser Leu Gln Phe Gln Gly Tyr
100 105 110
His Leu Cys Gly Gly Ser Val Ile Thr Pro Leu Trp Ile Ile Thr Ala
115 120 125
Ala His Cys Val Tyr Asp Leu Tyr Leu Pro Lys Ser Trp Thr Ile Gln
130 135 140
Val Gly Leu Val Ser Leu Leu Asp Asn Pro Ala Pro Ser His Leu Val
145 150 155 160
Glu Lys Ile Val Tyr His Ser Lys Tyr Lys Pro Lys Arg Leu Gly Asn
165 170 175
Asp Ile Ala Leu Met Lys Leu Ala Gly Pro Leu Thr Phe Asn Glu Met
180 185 190
Ile Gln Pro Val Cys Leu Pro Asn Ser Glu Glu Asn Phe Pro Asp Gly
195 200 205
Lys Val Cys Trp Thr Ser Gly Trp Gly Ala Thr Glu Asp Gly Ala Gly
210 215 220
Asp Ala Ser Pro Val Leu Asn His Ala Ala Val Pro Leu Ile Ser Asn
225 230 235 240
Lys Ile Cys Asn His Arg Asp Val Tyr Gly Gly Ile Ile Ser Pro Ser
245 250 255
Met Leu Cys Ala Gly Tyr Leu Thr Gly Gly Val Asp Ser Cys Gln Gly
260 265 270
Asp Ser Gly Gly Pro Leu Val Cys Gln Glu Arg Arg Leu Trp Lys Leu
275 280 285
Val Gly Ala Thr Ser Phe Gly Ile Gly Cys Ala Glu Val Asn Lys Pro
290 295 300
Gly Val Tyr Thr Arg Val Thr Ser Phe Leu Asp Trp Ile His Glu Gln
305 310 315 320
Met Glu Arg Asp Leu Lys Thr
325
<210>85
<211>1359
<212>DNA
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 3 transcript variant D
<400>85
accgggcacc ggacggctcg ggtactttcg ttcttaatta ggtcatgccc gtgtgagcca 60
ggaaagggct gtgtttatgg gaagccagta acactgtggc ctactatctc ttccgtggtg 120
ccatctacat ttttgggact cgggaattat gaggtagagg tggaggcgga gccggatgtc 180
agaggtcctg aaatagtcac catgggggaa aatgatccgc ctgctgttga agcccccttc 240
tcattccgat cgctttttgg ccttgatgat ttgaaaataa gtcctgttgc accagatgca 300
gatgctgttg ctgcacagat cctgtcactg ctgccattga agttttttcc aatcatcgtc 360
attgggatca ttgcattgat attagcactg gccattggtc tgggcatcca cttcgactgc 420
tcagggaagt acagatgtcg ctcatccttt aagtgtatcg agctgatagc tcgatgtgac 480
ggagtctcgg attgcaaaga cggggaggac gagtaccgct gtgtccgggt gggtggtcag 540
aatgccgtgc tccaggtgtt cacagctgct tcgtggaaga ccatgtgctc cgatgactgg 600
aagggtcact acgcaaatgt tgcctgtgcc caactgggtt tcccaagcta tgtgagttca 660
gataacctca gagtgagctc gctggagggg cagttccggg aggagtttgt gtccatcgat 720
cacctcttgc cagatgacaa ggtgactgca ttacaccact cagtatatgt gagggaggga 780
tgtgcctctg gccacgtggt taccttgcag tgcacagcct gtggtcatag aaggggctac 840
agctcacgca tcgtgggtgg aaacatgtcc ttgctctcgc agtggccctg gcaggccagc 900
cttcagttcc agggctacca cctgtgcggg ggctctgtca tcacgcccct gtggatcatc 960
actgctgcac actgtgttta tgacttgtac ctccccaagt catggaccat ccaggtgggt 1020
ctagtttccc tgttggacaa tccagcccca tcccacttgg tggagaagat tgtctaccac 1080
agcaagtaca agccaaagag gctgggcaat gacatcgccc ttatgaagct ggccgggcca 1140
ctcacgttca atggtacatc tgggtctcta tgtggttctg cagctcttcc tttgtttcaa 1200
gaggatttgc aattgctcat tgaagcattc ttatgatggc tgctttataa tccttgtcag 1260
atattaataa ttccaactcc tgattcatgt tggtgttggc atcagttgat tatcttttct 1320
cattaaaatt gtgatgctcc taaaaaaaaa aaaaaaaaa 1359
<210>86
<211>344
<212>PRT
<213>Homo sapiens
<220>
<223>transmembrane protease，serine 3 isoform 3
<400>86
Met Gly Glu Asn Asp Pro Pro Ala Val Glu Ala Pro Phe Ser Phe Arg
1 5 10 15
Ser Leu Phe Gly Leu Asp Asp Leu Lys Ile Ser Pro Val Ala Pro Asp
20 25 30
Ala Asp Ala Val Ala Ala Gln Ile Leu Ser Leu Leu Pro Leu Lys Phe
35 40 45
Phe Pro Ile Ile Val Ile Gly Ile Ile Ala Leu Ile Leu Ala Leu Ala
50 55 60
Ile Gly Leu Gly Ile His Phe Asp Cys Ser Gly Lys Tyr Arg Cys Arg
65 70 75 80
Ser Ser Phe Lys Cys Ile Glu Leu Ile Ala Arg Cys Asp Gly Val Ser
85 90 95
Asp Cys Lys Asp Gly Glu Asp Glu Tyr Arg Cys Val Arg Val Gly Gly
100 105 110
Gln Asn Ala Val Leu Gln Val Phe Thr Ala Ala Ser Trp Lys Thr Met
115 120 125
Cys Ser Asp Asp Trp Lys Gly His Tyr Ala Asn Val Ala Cys Ala Gln
130 135 140
Leu Gly Phe Pro Ser Tyr Val Ser Ser Asp Asn Leu Arg Val Ser Ser
145 150 155 160
Leu Glu Gly Gln Phe Arg Glu Glu Phe Val Ser Ile Asp His Leu Leu
165 170 175
Pro Asp Asp Lys Val Thr Ala Leu His His Ser Val Tyr Val Arg Glu
180 185 190
Gly Cys Ala Ser Gly His Val Val Thr Leu Gln Cys Thr Ala Cys Gly
195 200 205
His Arg Arg Gly Tyr Ser Ser Arg Ile Val Gly Gly Asn Met Ser Leu
210 215 220
Leu Ser Gln Trp Pro Trp Gln Ala Ser Leu Gln Phe Gln Gly Tyr His
225 230 235 240
Leu Cys Gly Gly Ser Val Ile Thr Pro Leu Trp Ile Ile Thr Ala Ala
245 250 255
His Cys Val Tyr Asp Leu Tyr Leu Pro Lys Ser Trp Thr Ile Gln Val
260 265 270
Gly Leu Val Ser Leu Leu Asp Asn Pro Ala Pro Ser His Leu Val Glu
275 280 285
Lys Ile Val Tyr His Ser Lys Tyr Lys Pro Lys Arg Leu Gly Asn Asp
290 295 300
Ile Ala Leu Met Lys Leu Ala Gly Pro Leu Thr Phe Asn Gly Thr Ser
305 310 315 320
Gly Ser Leu Cys Gly Ser Ala Ala Leu Pro Leu Phe Gln Glu Asp Leu
325 330 335
Gln Leu Leu Ile Glu Ala Phe Leu
340
<210>87
<211>1191
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK15 transcript variant 1
<400>87
actgcagccg aaccctggtc cctcctccac aatgtggctt ctcctcactc tctccttcct 60
gctggcatcc acagcagccc aggatggtga caagttgctg gaaggtgacg agtgtgcacc 120
ccactcccag ccatggcaag tggctctcta cgagcgtgga cgctttaact gtggcgcttc 180
cctcatctcc ccacactggg tgctgtctgc ggcccactgc caaagccgct tcatgagagt 240
gcgcctggga gagcacaacc tgcgcaagcg cgatggccca gagcaactac ggaccacgtc 300
tcgggtcatt ccacacccgc gctacgaagc gcgcagccac cgcaacgaca tcatgttgct 360
gcgcctagtc cagcccgcac gcctgaaccc ccagtgagtc tcccagatac gttgcattgt 420
gccaacatca gcattatctc ggacacatct tgtgacaaga gctacccagg gcgcctgaca 480
aacaccatgg tgtgtgcagg cgcggagggc agaggcgcag aatcctgtga gggtgactct 540
gggggacccc tggtctgtgg gggcatcctg cagggcattg tgtcctgggg tgacgtccct 600
tgtgacaaca ccaccaagcc tggtgtctat accaaagtct gccactactt ggagtggatc 660
agggaaacca tgaagaggaa ctgactattc tagcctatct cctgtgcccc tgactgagca 720
gaagccccca cagctggcca gcagccccgc ctgacatgga acagaacgga gccatccccc 780
aagaccctgt ccaaggccca gatgttagcc aaggacttgt cccacctgag gacaaagctg 840
gcgctcaagg tcacctgttt aatgccaaga taacaaagcg ctgatccaag ttgctctgta 900
ggaatttctg tgactttttt ctggggtcaa agagaaaccc cgagacactg tacactgttc 960
cttttcaccc accaccccga tccctaggtg aggagaagcg gcttgaagca gggctccatt 1020
cattcaacac acatgaccac ccgtgtgatc ttgaacaaga ggcccaatct cacttcgcct 1080
tggtttcctt atctgtaaaa tgagaccatc ttattgctga cttcaaaggg ctgttgtgag 1140
gattaaatga gatgattcgt ctgaactgat taaaatcgtg tctggcactg a 1191
<210>88
<211>121
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hK15 isoform 1
<400>88
Met Trp Leu Leu Leu Thr Leu Ser Phe Leu Leu Ala Ser Thr Ala Ala
1 5 10 15
Gln Asp Gly Asp Lys Leu Leu Glu Gly Asp Glu Cys Ala Pro His Ser
20 25 30
Gln Pro Trp Gln Val Ala Leu Tyr Glu Arg Gly Arg Phe Asn Cys Gly
35 40 45
Ala Ser Leu Ile Ser Pro His Trp Val Leu Ser Ala Ala His Cys Gln
50 55 60
Ser Arg Phe Met Arg ValArg Leu Gly Glu His Asn Leu Arg Lys Arg
65 70 75 80
Asp Gly Pro Glu Gln Leu Arg Thr Thr Ser Arg Val Ile Pro His Pro
85 90 95
Arg Tyr Glu Ala Arg Ser His Arg Asn Asp Ile Met Leu Leu Arg Leu
100 105 110
ValGln Pro Ala Arg Leu Asn Pro Gln
115 120
<210>89
<211>1172
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK15 transcript variant 2
<400>89
actgcagccg aaccctggtc cctcctccac aatgtggctt ctcctcactc tctccttcct 60
gctggcatcc acagcagccc aggatggtga caagttgctg gaaggtgacg agtgtgcacc 120
ccactcccag ccatggcaag tggctctcta cgagcgtgga cgctttaact gtggcgcttc 180
cctcatctcc ccacactggg tgctgtctgc ggcccactgc caaagccgct tcatgagagt 240
gcgcctggga gagcacaacc tgcgcaagcg cgatggccca gagcaactac ggaccacgtc 300
tcgggtcatt ccacacccgc gctacgaagc gcgcagccac cgcaacgaca tcatgttgct 360
gcgcctagtc cagcccgcac gcctgaaccc ccaggtgcgc cccgcggtgc tacccacgcg 420
ttgcccccac ccgggggagg cctgtgtggt gtctggctgg ggcctggtgt cccacaacga 480
gcctgggacc gctgggagcc cccggtcaca agggtgactc tgggggaccc ctggtctgtg 540
ggggcatcct gcagggcatt gtgtcctggg gtgacgtccc ttgtgacaac accaccaagc 600
ctggtgtcta taccaaagtc tgccactact tggagtggat cagggaaacc atgaagagga 660
actgactatt ctagcctatc tcctgtgccc ctgactgagc agaagccccc acagctggcc 720
agcagccccg cctgacatgg aacagaacgg agccatcccc caagaccctg tccaaggccc 780
agatgttagc caaggacttg tcccacctga ggacaaagct ggcgctcaag gtcacctgtt 840
taatgccaag ataaeaaagc getgatccaa gttgctctgt aggaatttct gtgacttttt 900
tctggggtca aagagaaacc ccgagacact gtacactgtt ccttttcacc caccaccccg 960
atccctaggt gaggagaagc ggcttgaage agggctccat tcattcaaca cacatgacca 1020
cccgtgtgat cttgaacaag aggcccaatc tcacttcgcc ttggtttcct tatctgtaaa 1080
atgagaccat cttattgctg acttcaaagg getgttgtga ggattaaatg agatgattcg 1140
tctgaactga ttaaaatcgt gtctggcact ga 1172
<210>90
<211>161
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hK15 isoform 2
<400>90
Met Trp Leu Leu Leu Thr Leu Ser Phe Leu Leu Ala Ser Thr Ala Ala
1 5 10 15
Gln Asp Gly Asp Lys Leu Leu Glu Gly Asp Glu Cys Ala Pro His Ser
20 25 30
Gln Pro Trp Gln Val Ala Leu Tyr Glu Arg Gly Arg Phe Asn Cys Gly
35 40 45
Ala Ser Leu Ile Ser Pro His Trp Val Leu Ser Ala Ala His Cys Gln
50 55 60
Ser Arg Phe Met Arg Val Arg Leu Gly Glu His Asn Leu Arg Lys Arg
65 70 75 80
Asp Gly Pro Glu Gln Leu Arg Thr Thr Ser Arg Val Ile Pro His Pro
85 90 95
Arg Tyr Glu Ala Arg Ser His Arg Asn Asp Ile Met Leu Leu Arg Leu
100 105 110
Val Gln Pro Ala Arg Leu Asn Pro Gln Val Arg Pro Ala Val Leu Pro
115 120 125
Thr Arg Cys Pro His Pro Gly Glu Ala Cys Val Val Ser Gly Trp Gly
130 135 140
Leu Val Ser His Asn Glu Pro Gly Thr Ala Gly Ser Pro Arg Ser Gln
145 150 155 160
Gly
<210>91
<211>1054
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK15 transcript variant 3
<400>91
actgcagccg aaccctggtc cctcctccac aatgtggctt ctcctcactc tctccttcct 60
gctggcatcc acagcagccc aggatggtga caagttgctg gaaggtgacg agtgtgcacc 120
ccactcccag ccatggcaag tggctctcta cgagcgtgga cgctttaact gtggcgcttc 180
cctcatctcc ccacactggg tgctgtctgc ggcccactgc caaagccgct tcatgagagt 240
gcgcctggga gagcacaacc tgcgcaagcg cgatggccca gagcaactac ggaccacgtc 300
tcgggtcatt ccacacccgc gctacgaagc gcgcagccac cgcaacgaca tcatgttgct 360
gcgcctagtc cagcccgcac gcctgaaccc ccagggtgac tctgggggac ccctggtctg 420
tgggggcatc ctgcagggca ttgtgtcctg gggtgacgtc ccttgtgaca acaccaccaa 480
gcctggtgtc tataccaaag tctgccacta cttggagtgg atcagggaaa ccatgaagag 540
gaactgacta ttctagccta tctcctgtgc ccctgactga gcagaagccc ccacagctgg 600
ccagcagccc cgcctgacat ggaacagaac ggagccatcc cccaagaccc tgtccaaggc 660
ccagatgtta gccaaggact tgtcccacct gaggacaaag ctggcgctca aggtcacctg 720
tttaatgcca agataacaaa gcgctgatcc aagttgctct gtaggaattt ctgtgacttt 780
tttctggggt caaagagaaa ccccgagaca ctgtacactg ttccttttca cccaccaccc 840
cgatccctag gtgaggagaa gcggcttgaa gcagggctcc attcattcaa cacacatgac 900
cacccgtgtg atcttgaaca agaggcccaa tctcacttcg ccttggtttc cttatctgta 960
aaatgagacc atcttattgc tgacttcaaa gggctgttgt gaggattaaa tgagatgatt 1020
cgtctgaact gattaaaatc gtgtctggca ctga 1054
<210>92
<211>171
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hK15 isoform 3
<400>92
Met Trp Leu Leu Leu Thr Leu Ser Phe Leu Leu Ala Ser Thr Ala Ala
1 5 10 15
Gln Asp Gly Asp Lys Leu Leu Glu Gly Asp Glu Cys Ala Pro His Ser
20 25 30
Gln Pro Trp Gln Val Ala Leu Tyr Glu Arg Gly Arg Phe Asn Cys Gly
35 40 45
Ala Ser Leu Ile Ser Pro His Trp Val Leu Ser Ala Ala His Cys Gln
50 55 60
Ser Arg Phe Met Arg Val Arg Leu Gly Glu His Asn Leu Arg Lys Arg
65 70 75 80
Asp Gly Pro Glu Gln Leu Arg Thr Thr Ser Arg Val Ile Pro His Pro
85 90 95
Arg Tyr Glu Ala Arg Ser His Arg Asn Asp Ile Met Leu Leu Arg Leu
100 105 110
Val Gln Pro Ala Arg Leu Asn Pro Gln Gly Asp Ser Gly Gly Pro Leu
115 120 125
Val Cys Gly Gly Ile Leu Gln Gly Ile Val Ser Trp Gly Asp Val Pro
130 135 140
Cys Asp Asn Thr Thr Lys Pro Gly Val Tyr Thr Lys Val Cys His Tyr
145 150 155 160
Leu Glu Trp Ile Arg Glu Thr Met Lys Arg Asn
165 170
<210>93
<211>1309
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK15 transcript variant 4
<400>93
actgcagccg aaccctggtc cctcctccac aatgtggctt ctcctcactc tctccttcct 60
gctggcatcc acagcagccc aggatggtga caagttgctg gaaggtgacg agtgtgcacc 120
ccactcccag ccatggcaag tggctctcta cgagcgtgga cgctttaact gtggcgcttc 180
cctcatctcc ccacactggg tgctgtctgc ggcccactgc caaagccgct tcatgagagt 240
gcgcctggga gagcacaacc tgcgcaagcg cgatggccca gagcaactac ggaccacgtc 300
tcgggtcatt ccacacccgc gctacgaagc gcgcagccac cgcaacgaca tcatgttgct 360
gcgcctagtc cagcccgcac gcctgaaccc ccaggtgcgc cccgcggtgc tacccacgcg 420
ttgcccccac ccgggggagg cctgtgtggt gtctggctgg ggcctggtgt cccacaacga 480
gcctgggacc gctgggagcc cccggtcaca agtgagtctc ccagatacgt tgcattgtgc 540
caacatcagc attatctcgg acacatcttg tgacaagagc tacccagggc gcctgacaaa 600
caccatggtg tgtgcaggcg cggagggcag aggcgcagaa tcctgtgagg gtgactctgg 660
gggacccctg gtctgtgggg gcatcctgca gggcattgtg tcctggggtg acgtcccttg 720
tgacaacacc accaagcctg gtgtctatac caaagtctgc cactacttgg agtggatcag 780
ggaaaccatg aagaggaact gactattcta gcctatctcc tgtgcccctg actgagcaga 840
agcccccaca gctggccagc agccccgcct gacatggaac agaacggagc catcccccaa 900
gaccctgtcc aaggcccaga tgttagccaa ggacttgtcc cacctgagga caaagctggc 960
gctcaaggtc acctgtttaa tgccaagata acaaagcgct gatccaagtt gctctgtagg 1020
aatttctgtg acttttttct ggggtcaaag agaaaccccg agacactgta cactgttcct 1080
tttcacccac caccccgatc cctaggtgag gagaagcggc ttgaagcagg gctccattca 1140
ttcaacacac atgaccaccc gtgtgatctt gaacaagagg cccaatctca cttcgccttg 1200
gtttccttat ctgtaaaatg agaccatctt attgctgact tcaaagggct gttgtgagga 1260
ttaaatgaga tgattcgtct gaactgatta aaatcgtgtc tggcactga 1309
<210>94
<211>256
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hK15 isoform 4
<400>94
Met Trp Leu Leu Leu Thr Leu Ser Phe Leu Leu Ala Ser Thr Ala Ala
1 5 10 15
Gln Asp Gly Asp Lys Leu Leu Glu Gly Asp Glu Cys Ala Pro His Ser
20 25 30
Gln Pro Trp Gln Val Ala Leu Tyr Glu Arg Gly Arg Phe Asn Cys Gly
35 40 45
Ala Ser Leu Ile Ser Pro His Trp Val Leu Ser Ala Ala His Cys Gln
50 55 60
Ser Arg Phe Met Arg Val Arg Leu Gly Glu His Asn Leu Arg Lys Arg
65 70 75 80
Asp Gly Pro Glu Gln Leu Arg Thr Thr Ser Arg Val Ile Pro His Pro
85 90 95
Arg Tyr Glu Ala Arg Ser His Arg Asn Asp Ile Met Leu Leu Arg Leu
100 105 110
Val Gln Pro Ala Arg Leu Asn Pro Gln Val Arg Pro Ala Val Leu Pro
115 120 125
Thr Arg Cys Pro His Pro Gly Glu Ala Cys Val Val Ser Gly Trp Gly
130 135 140
Leu Val Ser His Asn Glu Pro Gly Thr Ala Gly Ser Pro Arg Ser Gln
145 150 155 160
Val Ser Leu Pro Asp Thr Leu His Cys Ala Asn Ile Ser Ile Ile Ser
165 170 175
Asp Thr Ser Cys Asp Lys Ser Tyr Pro Gly Arg Leu Thr Asn Thr Met
180 185 190
Val Cys Ala Gly Ala Glu Gly Arg Gly Ala Glu Ser Cys Glu Gly Asp
195 200 205
Ser Gly Gly Pro Leu Val Cys Gly Gly Ile Leu Gln Gly Ile Val Ser
210 215 220
Trp Gly Asp Val Pro Cys Asp Asn Thr Thr Lys Pro Gly Val Tyr Thr
225 230 235 240
Lys Val Cys His Tyr Leu Glu Trp Ile Arg Glu Thr Met Lys Arg Asn
245 250 255
<210>95
<211>920
<212>DNA
<213>Homo sapiens
<220>
<223>Mername-AA031 peptidase
<400>95
cagacaggag gcgtctgtag agatatcatg aacttcaact tagctttgtt ttccagagac 60
tggagctaaa ctgggctttc aacatcatca tgaagtttat cctcctctgg gccctcttga 120
atctgactgt tgctttggcc tttaatccag attacacagt cagctccact cccccttact 180
tggtctattt gaaatctgac tacttgccct gcgctggagt cctgatccac ccgctttggg 240
tgatcacagc tgcacactgc aatttaccaa agcttcgggt gatattgggg gttacaatcc 300
cagcagactc taatgaaaag catctgcaag tgattggcta tgagaagatg attcatcatc 360
cacacttctc agtcacttct attgatcatg acatcatgct aatcaagctg aaaacagagg 420
ctgaactcaa tgactatgtg aaattagcca acctgcccta ccaaactatc tctgaaaata 480
ccatgtgctc tgtctctacc tggagctaca atgtgtgtga tatctacaaa gagcccgatt 540
cactgcaaac tgtgaacatc tctgtaatct ccaagcctca gtgtcgcgat gcctataaaa 600
cctacaacat cacggaaaat atgctgtgtg tgggcattgt gccaggaagg aggcagccct 660
gcaaggaagt ttctgctgcc ccggcaatct gcaatgggat gcttcaagga atcctgtctt 720
ttgcggatgg atgtgttttg agagccgatg ttggcatcta tgccaaaatt ttttactata 780
taccctggat tgaaaatgta atccaaaata actgagctgt ggcagttgtg gaccatatga 840
cacagcttgt ccccatcgtt cacctttaga attaaatata aattaactcc tcaaaaaaaa 900
aaaaaaaaaa aaaaaaaaaa 920
<210>96
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mername-AA031 peptidase
<400>96
Met Lys Phe Ile Leu Leu Trp Ala Leu Leu Asn Leu Thr Val Ala Leu
1 5 10 15
Ala Phe Asn Pro Asp Tyr Thr Val Ser Ser Thr Pro Pro Tyr Leu Val
20 25 30
Tyr Leu Lys Ser Asp Tyr Leu Pro Cys Ala Gly Val Leu Ile His Pro
35 40 45
Leu Trp Val Ile Thr Ala Ala His Cys Asn Leu Pro Lys Leu Arg Val
50 55 60
Ile Leu Gly ValThr Ile Pro Ala Asp Ser Asn Glu Lys His Leu Gln
65 70 75 80
Val Ile Gly Tyr Glu Lys Met Ile His His Pro His Phe Ser Val Thr
85 90 95
Ser Ile Asp His Asp Ile Met Leu Ile Lys Leu Lys Thr Glu Ala Glu
100 105 110
Leu Asn Asp Tyr Val Lys Leu Ala Asn Leu Pro Tyr Gln Thr Ile Ser
115 120 125
Glu Asn Thr Met Cys Ser Val Ser Thr Trp Ser Tyr Asn Val Cys Asp
130 135 140
Ile Tyr Lys Glu Pro Asp Ser Leu Gln Thr Val Asn Ile Ser Val Ile
145 150 155 160
Ser Lys Pro Gln Cys Arg Asp Ala Tyr Lys Thr Tyr Asn Ile Thr Glu
165 170 175
Asn Met Leu Cys Val Gly Ile Val Pro Gly Arg Arg Gln Pro Cys Lys
180 185 190
Glu Val Ser Ala Ala Pro Ala Ile Cys Asn Gly Met Leu Gln Gly Ile
195 200 205
Leu Ser Phe Ala Asp Gly Cys Val Leu Arg Ala Asp Val Gly Ile Tyr
210 215 220
Ala Lys Ile Phe Tyr Tyr Ile Pro Trp Ile Glu Asn Val Ile Gln Asn
225 230 235 240
Asn
<210>97
<211>2393
<212>DNA
<213>Homo sapiens
<220>
<223>membrane-type mosaic serine protease
<400>97
cgcccgggca ggttgagaag ccaggggcca agatggatct tctcctcgac atcagctaag 60
cctggaggac tcttcccctc agagaccatg gagagggaca gccacgggaa tgcatctcca 120
gcaagaacac cttcagctgg agcatctcca gcccaggcat ctccagctgg gacacctcca 180
ggccgggcat ctccagccca ggcatctcca gcccaggcat ctccagctgg gacacctccg 240
ggccgggcat ctccagccca ggcatctcca gctggtacac ctccaggccg ggcatctcca 300
ggccgggcat ctccagccca ggcatctcca gcccgggcat ctccggctct ggcatcactt 360
tccaggtcct catccggcag gtcatcatcc gccaggtcag cctcggtgac aacctcccca 420
accagagtgt accttgttag agcaacacca gtgggggctg tacccatccg atcatctcct 480
gccaggtcag caccagcaac cagggccacc agggagagcc caggtacgag cctgcccaag 540
ttcacctggc gggagggcca gaagcagcta ccgctcatcg ggtgcgtgct cctcctcatt 600
gccctggtgg tttcgctcat catcctcttc cagttctggc agggccacac agggatcagg 660
cacaaggagc agagggagag ctgtcccaag cacgctgttc gctgtgacgg ggtggtggac 720
tgcaagctga agagtgacga gctgggctgc gtgaggtttg actgggacaa gtctctgctt 780
aaaatctact ctgggtcctc ccatcagtgg cttcccatct gtagcagcaa ctggaatgac 840
tcctactcag agaagacctg ccagcagctg ggtttcgaga gtgctcaccg gacaaccgag 900
gttgcccaca gggattttgc caacagcttc tcaatcttga gatacaactc caccatccag 960
gaaagcctcc acaggtctga atgcccttcc cagcggtata tctccctcca gtgttcccac 1020
tgcggactga gggccatgac cgggcggatc gtgggagggg cgctggcctc ggatagcaag 1080
tggccttggc aagtgagtct gcacttcggc accacccaca tctgtggagg cacgctcatt 1140
gacgcccagt gggtgctcac tgccgcccac tgcttcttcg tgacccggga gaaggtcctg 1200
gagggctgga aggtgtacgc gggcaccagc aacctgcacc agttgcctga ggcagcctcc 1260
attgccgaga tcatcatcaa cagcaattac accgatgagg aggacgacta tgacatcgcc 1320
ctcatgcggc tgtccaagcc cctgaccctg tccgctcaca tccaccctgc ttgcctcccc 1380
atgcatggac agacctttag cctcaatgag acctgctgga tcacaggctt tggcaagacc 1440
agggagacag atgacaagac atcccccttc ctccgggagg tgcaggtcaa tctcatcgac 1500
ttcaagaaat gcaatgacta cttggtctat gacagttacc ttaccccaag gatgatgtgt 1560
gctggggacc ttcgtggggg cagagactcc tgccagggag acagcggggg gcctcttgtc 1620
tgtgagcaga acaaccgctg gtacctggca ggtgtcacca gctggggcac aggctgtggc 1680
cagagaaaca aacctggtgt gtacaccaaa gtgacagaag ttcttccctg gatttacagc 1740
aagatggagg taagatccct gcagcaggac actgcaccca gcaggctggg aacttcctca 1800
ggtggggacc ctggaggagc acccagggtg taggcagagg tcccctcagc gtccccatat 1860
tcggggggtg ttctggacag ggtcaaatgt gatgcctggg gtcaatccca gctgtctgtg 1920
tttctttccc tgcttttctt ccctcagaac agagctcagc gggttgaaaa agggtggacc 1980
tacaggccag gcaggcagtt gctgggcaga tgttctccca gaagtatttt tttgtgtaag 2040
gttgcaatgg actttgaaaa cgtttcagtt tctgcagagg attttgtgat agtctttgtt 2100
atcaagcatt tatgcatggg aatccgctct tcatggcctt tcccagctct gtttgtttta 2160
gtctttttga ttttcttttt gttgttgttg ttgtcttttt ttaaaaacac aagtgactcc 2220
attttaactc tgacaacttt cacagctgtc accagaatgc tccctgagaa ctaccattct 2280
ttccctttcc cacttaaaat atttcatcag aacctcacca ctatcataaa agagtataaa 2340
gtaataaaat aataaaaagc gaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaa 2393
<210>98
<211>581
<212>PRT
<213>Homo Sapiens
<220>
<223>membrane-type mosaic serine protease
<400>98
Met Glu Arg Asp Ser His Gly Asn Ala Ser Pro Ala Arg Thr Pro Ser
1 5 10 15
Ala Gly Ala Ser Pro Ala Gln Ala Ser Pro Ala Gly Thr Pro Pro Gly
20 25 30
Arg Ala Ser Pro Ala Gln Ala Ser Pro Ala Gln Ala Ser Pro Ala Gly
35 40 45
Thr Pro Pro Gly Arg Ala Ser Pro Ala Gln Ala Ser Pro Ala Gly Thr
50 55 60
Pro Pro Gly Arg Ala Ser Pro Gly Arg Ala Ser Pro Ala Gln Ala Ser
65 70 75 80
Pro Ala Arg Ala Ser Pro Ala Leu Ala Ser Leu Ser Arg Ser Ser Ser
85 90 95
Gly Arg Ser Ser Ser Ala Arg Ser Ala Ser Val Thr Thr Ser Pro Thr
100 105 110
Arg Val Tyr Leu Val Arg Ala Thr Pro Val Gly Ala Val Pro Ile Arg
115 120 125
Ser Ser Pro Ala Arg Ser Ala Pro Ala Thr Arg Ala Thr Arg Glu Ser
130 135 140
Pro Gly Thr Ser Leu Pro Lys Phe Thr Trp Arg Glu Gly Gln Lys Gln
145 150 155 160
Leu Pro Leu Ile Gly Cys Val Leu Leu Leu Ile Ala Leu Val Val Ser
165 170 175
Leu Ile Ile Leu Phe Gln Phe Trp Gln Gly His Thr Gly Ile Arg His
180 185 190
Lys Glu Gln Arg Glu Ser Cys Pro Lys His Ala Val Arg Cys Asp Gly
195 200 205
Val Val Asp Cys Lys Leu Lys Ser Asp Glu Leu Gly Cys Val Arg Phe
210 215 220
Asp Trp Asp Lys Ser Leu Leu Lys Ile Tyr Ser Gly Ser Ser His Gln
225 230 235 240
Trp Leu Pro Ile Cys Ser Ser Asn Trp Asn Asp Ser Tyr Ser Glu Lys
245 250 255
Thr Cys Arg Gln Leu Gly Phe Glu Ser Ala His Arg Thr Thr Glu Val
260 265 270
Ala His Arg Asp Phe Ala Asn Ser Phe Ser Ile Leu Arg Tyr Asn Ser
275 280 285
Thr Ile Gln Glu Ser Leu His Arg Ser His Cys Pro Ser Gln Arg Tyr
290 295 300
Ile Ser Leu Gln Cys Ser His Cys Gly Leu Arg Ala Met Thr Gly Arg
305 310 315 320
Ile Val Gly Gly Ala Leu Ala Ser Asp Ser Lys Trp Pro Trp Gln Val
325 330 335
Ser Leu His Phe Gly Thr Thr His Ile Cys Gly Gly Thr Leu Ile Asp
340 345 350
Ala Gln Trp Val Leu Thr Ala Ala His Cys Phe Phe Val Thr Arg Glu
355 360 365
Lys Val Leu Glu Gly Trp Lys Val Tyr Ala Gly Thr Ser Asn Leu His
370 375 380
Gln Leu Pro Glu Ala Ala Ser Ile Ala Glu Ile Ile Ile Asn Ser Asn
385 390 395 400
Tyr Thr Asp Glu Glu Asp Asp Tyr Asp Ile Ala Leu Met Arg Leu Ser
405 410 415
Lys Pro Leu Thr Leu Ser Ala His Ile His Pro Ala Cys Leu Pro Met
420 425 430
His Gly Gln Thr Phe Ser Leu Asn Glu Thr Cys Trp Ile Thr Gly Phe
435 440 445
Gly Lys Thr Arg Glu Thr Asp Asp Lys Thr Ser Pro Phe Leu Arg Glu
450 455 460
Val Gln Val Asn Leu Ile Asp Phe Lys Lys Cys Asn Asp Tyr Leu Val
465 470 475 480
Tyr Asp Ser Tyr Leu Thr Pro Arg Met Met Cys Ala Gly Asp Leu His
485 490 495
Gly Gly Arg Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys
500 505 510
Glu Gln Asn Asn Arg Trp Tyr Leu Ala Gly Val Thr Ser Trp Gly Thr
515 520 525
Gly Cys Gly Gln Arg Asn Lys Pro Gly Val Tyr Thr Lys Val Thr Glu
530 535 540
Val Leu Pro Trp Ile Tyr Ser Lys Met Glu Val Arg Ser Leu Gln Gln
545 550 555 560
Asp Thr Ala Pro Ser Arg Leu Gly Thr Ser Ser Gly Gly Asp Pro Gly
565 570 575
Gly Ala Pro Arg Val
580
<210>99
<211>148
<212>PRT
<213>Homo sapiens
<220>
<223>mername-AA038 peptidase
<400>99
Cys Gly Lys Pro Lys Val Val Gly Lys Ile Tyr Gly Gly Arg Asp Ala
1 5 10 15
Ala Ala Gly Gln Trp Pro Trp Gln Ala Ser Leu Leu Tyr Trp Gly Ser
20 25 30
His Leu Cys Gly Ala Val Leu Ile Asp Ser Cys Trp Leu Val Ser Thr
35 40 45
Thr His Cys Phe Leu Asn Lys Ser Gln Ala Pro Lys Asn Tyr Gln Val
50 55 60
Leu Leu Gly Asn Ile Gln Leu Tyr His Gln Thr Gln His Thr Gln Lys
65 70 75 80
Met Ser Val His Arg Ile Ile Thr His Pro Asp Phe Glu Lys Leu His
85 90 95
Pro Phe Gly Ser Asp Ile Ala Met Leu Gln Leu His Leu Pro Met Asn
100 105 110
Phe Thr Ser Tyr Ile Val Pro Val Cys Leu Pro Ser Arg Asp Met Gln
115 120 125
Leu Pro Ser Asn Val Ser Cys Trp Ile Thr Gly Trp Gly Met Ala Ile
130 135 140
Leu Gly Gly Leu
145
<210>100
<211>712
<212>DNA
<213>Homo sapiens
<220>
<223>mername-AA128 peptidase
<400>100
cacagagggc tccagcgtga ttgccactgg ttaatgacct gccacccccg cccccccaac 60
ctctggtgcc gtccaaggac tctgacccca gtgcagtgtc tgtagggaag aggagccatg 120
gggcttcggg caggccccat cctgcttctg ctgctgtggc tgctgccaga atgcggccac 180
tccaaggagg ccgggaggat tgtgggaggc caagacaccc aggaaggacg ctggccgtgg 240
caggttggcc tgtggttgac ctcagtgggg catgtatgtg ggggcttcct catccaccca 300
cgctgggtgc tcacagccgc ccactgcttc ctgagggtga ctccgggggg ccgctggtct 360
gccccatcaa tgatacgtgg atccaggccg gcattgtgag ctggggattc ggctgtgccc 420
ggcctttccg gcctggtgtc tacacccagg tgctaagcta cacagactgg attcagagaa 480
ccctggctga atctcactca ggcatgtctg gggcccgccc aggtgcccca ggatcccact 540
caggcacctc cagatcccac ccagtgctgc tgcttgagct gttgaccgta tgcttgcttg 600
ggtccctgtg aaccatgagc catggagtcc gggatcccct ttctggtagg attgatggaa 660
tctaataata aaaactgtag gttttttatg tgaaaaaaaa aaaaaaaaaa aa 712
<210>101
<211>146
<212>PRT
<213>Homo sapiens
<220>
<223>mername-AA128 peptidase
<400>101
Met Arg Pro Leu Gln Gly Gly Arg Glu Asp Cys Gly Arg Pro Arg His
1 5 10 15
Pro Gly Arg Thr Leu Ala Val Ala Gly Trp Pro Val Val Asp Leu Ser
20 25 30
Gly Ala Cys Met Trp Gly Leu Pro His Pro Pro Thr Leu Gly Ala His
35 40 45
Ser Arg Pro Leu Leu Pro Glu Gly Asp Ser Gly Gly Pro Leu ValCys
50 55 60
Pro Ile Asn Asp Thr Trp Ile Gln Ala Gly Ile Val Ser Trp Gly Phe
65 70 75 80
Gly Cys Ala Arg Pro Phe Arg Pro Gly Val Tyr Thr Gln Val Leu Ser
85 90 95
Tyr Thr Asp Trp Ile Gln Arg Thr Leu Ala Glu Ser His Ser Gly Met
100 105 110
Ser Gly Ala Arg Pro Gly Ala Pro Gly Ser His Ser Gly Thr Ser Arg
115 120 125
Ser His Pro Val Leu Leu Leu Glu Leu Leu Thr Val Cys Leu Leu Gly
130 135 140
Ser Leu
145
<210>102
<211>800
<212>DNA
<213>Homo sapiens
<220>
<223>cationic trypsin(Homo sapiens-type 1)(cationic)
<400>102
accaccatga atccactcct gatccttacc tttgtggcag ctgctcttgc tgcccccttt 60
gatgatgatg acaagatcgt tgggggctac aactgtgagg agaattctgt cccctaccag 120
gtgtccctga attctggcta ccacttctgt ggtggctccc tcatcaacga acagtgggtg 180
gtatcagcag gccactgcta caagtcccgc atccaggtga gactgggaga gcacaacatc 240
gaagtcctgg aggggaatga gcagttcatc aatgcagcca agatcatccg ccacccccaa 300
tacgacagga agactctgaa caatgacatc atgttaatca agctctcctc acgtgcagta 360
atcaacgccc gcgtgtccac catctctctg cccaccgccc ctccagccac tggcacgaag 420
tgcctcatct ctggctgggg caacactgcg agctctggcg ccgactaccc agacgagctg 480
cagtgcctgg acgctcctgt gctgagccag gctaagtgtg aagcctccta ccctggaaag 540
attaccagca acatgttctg tgtgggcttc cttgagggag gcaaggattc atgtcagggt 600
gattctggtg gccctgtggt ctgcaatgga cagctccaag gagttgtctc ctggggtgat 660
ggctgtgccc agaagaacaa gcctggagtc tacaccaagg tctacaacta tgtgaaatgg 720
attaagaaca ccatagctgc caatagctaa agcccccagt atctcttcag tctctatacc 780
aataaagtga ccctgttctc 800
<210>103
<211>247
<212>PRT
<213>Homo sapiens
<220>
<223>cationic trypsin(Homo sapiens-type 1)(cationic)
<400>103
Met Asn Pro Leu Leu Ile Leu Thr Phe Val Ala Ala Ala Leu Ala Ala
1 5 10 15
Pro Phe Asp Asp Asp Asp Lys Ile Val Gly Gly Tyr Asn Cys Glu Glu
20 25 30
Asn Ser Val Pro Tyr Gln Val Ser Leu Asn Ser Gly Tyr His Phe Cys
35 40 45
Gly Gly Ser Leu Ile Asn Glu Gln Trp Val Val Ser Ala Gly His Cys
50 55 60
Tyr Lys Ser Arg Ile Gln Val Arg Leu Gly Glu His Asn Ile Glu Val
65 70 75 80
Leu Glu Gly Asn Glu Gln Phe Ile Asn Ala Ala Lys Ile Ile Arg His
85 90 95
Pro Gln Tyr Asp Arg Lys Thr Leu Asn Asn Asp Ile Met Leu Ile Lys
100 105 110
Leu Ser Ser Arg Ala Val Ile Asn Ala Arg Val Ser Thr Ile Ser Leu
115 120 125
Pro Thr Ala Pro Pro Ala Thr Gly Thr Lys Cys Leu Ile Ser Gly Trp
130 135 140
Gly Asn Thr Ala Ser Ser Gly Ala Asp Tyr Pro Asp Glu Leu Gln Cys
145 150 155 160
Leu Asp Ala Pro Val Leu Ser Gln Ala Lys Cys Glu Ala Ser Tyr Pro
165 170 175
Gly Lys Ile Thr Ser Asn Met Phe Cys Val Gly Phe Leu Glu Gly Gly
180 185 190
Lys Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Val Val Cys Asn Gly
195 200 205
Gln Leu Gln Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln Lys Asn
210 215 220
Lys Pro Gly Val Tyr Thr Lys Val Tyr Asn Tyr Val Lys Trp Ile Lys
225 230 235 240
Asn Thr Ile Ala Ala Asn Ser
245
<210>104
<211>938
<212>DNA
<213>Homo sapiens
<220>
<223>Neutrophil elastase
<400>104
gcacggaggg gcagagaccc cggagcccca gccccaccat gaccctcggc cgccgactcg 60
cgtgtctttt cctcgcctgt gtcctgccgg ccttgctgct ggggggcacc gcgctggcct 120
cggagattgt ggggggccgg cgagcgcggc cccacgcgtg gcccttcatg gtgtccctgc 180
agctgcgcgg aggccacttc tgcggcgcca ccctgattgc gcccaacttc gtcatgtcgg 240
ccgcgcactg cgtggcgaat gtaaacgtcc gcgcggtgcg ggtggtcctg ggagcccata 300
acctctcgcg gcgggagccc acccggcagg tgttcgccgt gcagcgcatc ttcgaaaacg 360
gctacgaccc cgtaaacttg ctcaacgaca tcgtgattct ccagctcaac gggtcggcca 420
ccatcaacgc caacgtgcag gtggcccagc tgccggctca gggacgccgc ctgggcaacg 480
gggtgcagtg cctggccatg ggctggggcc ttctgggcag gaaccgtggg atcgccagcg 540
tcctgcagga gctcaacgtg acggtggtga cgtccctctg ccgtcgcagc aacgtctgca 600
ctctcgtgag gggccggcag gccggcgtct gtttcgggga ctccggcagc cccttggtct 660
gcaacgggct aatccacgga attgcctcct tcgtccgggg aggctgcgcc tcagggctct 720
accccgatgc ctttgccccg gtggcacagt ttgtaaactg gatcgactct atcatccaac 780
gctccgagga caacccctgt ccccaccccc gggacccgga cccggccagc aggacccact 840
gagaagggct gcccgggtca cctcagctgc ccacacccac actctccagc atctggcaca 900
ataaacattc tctgttttgt agaaaaaaaa aaaaaaaa 938
<210>105
<211>267
<212>PRT
<213>Homo sapiens
<220>
<223>Neutrophil elastase
<400>105
Met Thr Leu Gly Arg Arg Leu Ala Cys Leu Phe Leu Ala Cys Val Leu
1 5 10 15
Pro Ala Leu Leu Leu Gly Gly Thr Ala Leu Ala Ser Glu Ile Val Gly
20 25 30
Gly Arg Arg Ala Arg Pro His Ala Trp Pro Phe Met Val Ser Leu Gln
35 40 45
Leu Arg Gly Gly His Phe Cys Gly Ala Thr Leu Ile Ala Pro Asn Phe
50 55 60
Val Met Ser Ala Ala His Cys Val Ala Asn Val Asn Val Arg Ala Val
65 70 75 80
Arg Val Val Leu Gly Ala His Asn Leu Ser Arg Arg Glu Pro Thr Arg
85 90 95
Gln Val Phe Ala Val Gln Arg Ile Phe Glu Asn Gly Tyr Asp Pro Val
100 105 110
Asn Leu Leu Asn Asp Ile Val Ile Leu Gln Leu Asn Gly Ser Ala Thr
115 120 125
Ile Asn Ala Asn Val Gln Val Ala Gln Leu Pro Ala Gln Gly Arg Arg
130 135 140
Leu Gly Asn Gly Val Gln Cys Leu Ala Met Gly Trp Gly Leu Leu Gly
145 150 155 160
Arg Asn Arg Gly Ile Ala Ser Val Leu Gln Glu Leu Asn Val Thr Val
165 170 175
ValThr Ser Leu Cys Arg Arg Ser Asn Val Cys Thr Leu ValArg Gly
180 185 190
Arg Gln Ala Gly Val Cys Phe Gly Asp Ser Gly Ser Pro Leu Val Cys
195 200 205
Asn Gly Leu Ile His Gly Ile Ala Ser Phe Val Arg Gly Gly Cys Ala
210 215 220
Ser Gly Leu Tyr Pro Asp Ala Phe Ala Pro Val Ala Gln Phe Val Asn
225 230 235 240
Trp Ile Asp Ser Ile Ile Gln Arg Ser Glu Asp Asn Pro Cys Pro His
245 250 255
Pro Arg Asp Pro Asp Pro Ala Ser Arg Thr His
260 265
<210>106
<211>3895
<212>DNA
<213>Homo sapiens
<220>
<223>mannan-binding lectin-associated serine protease-3
<400>106
attccggcac agggacacaa acaagctcac ccaacaaagc caagctggga ggaccaaggc 60
cgggcagccg ggagcaccca aggcaggaaa atgaggtggc tgcttctcta ttatgctctg 120
tgcttctccc tgtcaaaggc ttcagcccac accgtggagc taaacaatat gtttggccag 180
atccagtcgc ctggttatcc agactcctat cccagtgatt cagaggtgac ttggaatatc 240
actgtcccag atgggtttcg gatcaagctt tacttcatgc acttcaactt ggaatcctcc 300
tacctttgtg aatatgacta tgtgaaggta gaaactgagg accaggtgct ggcaaccttc 360
tgtggcaggg agaccacaga cacagagcag actcccggcc aggaggtggt cctctcccct 420
ggctccttca tgtccatcac tttccggtca gatttctcca atgaggagcg tttcacaggc 480
tttgatgccc actacatggc tgtggatgtg gacgagtgca aggagaggga ggacgaggag 540
ctgtcctgtg accactactg ccacaactac attggcggct actactgctc ctgccgcttc 600
ggctacatcc tccacacaga caacaggacc tgccgagtgg agtgcagtga caacctcttc 660
actcaaagga ctggggtgat caccagccct gacttcccaa acccttaccc caagagctct 720
gaatgcctgt ataccatcga gctggaggag ggtttcatgg tcaacctgca gtttgaggac 780
atatttgaca ttgaggacca tcctgaggtg ccctgcccct atgactacat caagatcaaa 840
gttggtccaa aagttttggg gcctttctgt ggagagaaag ccccagaacc catcagcacc 900
cagagccaca gtgtcctgat cctgttccat agtgacaact cgggagagaa ccggggctgg 960
aggctctcat acagggctgc aggaaatgag tgcccagagc tacagcctcc tgtccatggg 1020
aaaatcgagc cctcccaagc caagtatttc ttcaaagacc aagtgctcgt cagctgtgac 1080
acaggctaca aagtgctgaa ggataatgtg gagatggaca cattccagat tgagtgtctg 1140
aaggatggga cgtggagtaa caagattccc acctgtaaaa ttgtagactg tagagcccca 1200
ggagagctgg aacacgggct gatcaccttc tctacaagga acaacctcac cacatacaag 1260
tctgagatca aatactcctg tcaggagccc tattacaaga tgctcaacaa taacacaggt 1320
atatatacct gttctgccca aggagtctgg atgaataaag tattggggag aagcctaccc 1380
acctgccttc cagagtgtgg tcagccctcc cgctccctgc caagcctggt caagaggatc 1440
attgggggcc gaaatgctga gcctggcctc ttcccgtggc aggccctgat agtggtggag 1500
gacacttcga gagtgccaaa tgacaagtgg tttgggagtg gggccctgct ctctgcgtcc 1560
tggatcctca cagcagctca tgtgctgcgc tcccagcgta gagacaccac ggtgatacca 1620
gtctccaagg agcatgtcac cgtctacctg ggcttgcatg atgtgcgaga caaatcgggg 1680
gcagtcaaca gctcagctgc ccgagtggtg ctccacccag acttcaacat ccaaaactac 1740
aaccacgata tagctctggt gcagctgcag gagcctgtgc ccctgggacc ccacgttatg 1800
cctgtctgcc tgccaaggct tgagcctgaa ggcccggccc cccacatgct gggcctggtg 1860
gccggctggg gcatctccaa tcccaatgtg acagtggatg agatcatcag cagtggcaca 1920
cggaccttgt cagatgtcct gcagtatgtc aagttacccg tggtgcctca cgctgagtgc 1980
aaaactagct atgagtcccg ctcgggcaat tacagcgtca cggagaacat gttctgtgct 2040
ggctactacg agggcggcaa agacacgtgc cttggagata gcggtggggc ctttgtcatc 2100
tttgatgact tgagccagcg ctgggtggtg caaggcctgg tgtcctgggg gggacctgaa 2160
gaatgcggca gcaagcaggt ctatggagtc tacacaaagg tctccaatta cgtggactgg 2220
gtgtgggagc agatgggctt accacaaagt gttgtggagc cccaggtgga acggtgagct 2280
gacttacttc ctcggggcct gcctcccctg agcgaagcta caccgcactt ccgacagcac 2340
actccacatt acttatcaga ccatatggaa tggaacacac tgacctagcg gtggcttctc 2400
ctaccgagac agcccccagg accctgagag gcagagtgtg gtatagggaa aaggctccag 2460
gcaggagacc tgtgttcctg agcttgtcca agtctctttc cctgtctggg cctcactcta 2520
ccgagtaata caatgcagga gctcaaccaa ggcctctgtg ccaatcccag cactcctttc 2580
caggccatgc ttcttacccc agtggccttt attcactcct gaccacttat caaacccatc 2640
ggtcctactg ttggtataac tgagcttgga cctgactatt agaaaatggt ttctaacatt 2700
gaactgaatg ccgcatctgt atattttcct gctctgcctt ctgggactag ccttggccta 2760
atccttcctc taggagaaga gcattcaggt tttgggagat ggctcatagc caagcccctc 2820
tctcttagtg tgatcccttg gagcaccttc atgcctgggg tttctctccc aaaagcttct 2880
tgcagtctaa gccttatccc ttatgttccc cattaaagga atttcaaaag acatggagaa 2940
agttgggaag gtttgtgctg actgctggga gcagaatagc cgtgggaggc ccaccaagcc 3000
cttaaattcc cattgtcaac tcagaacaca tttgggccca tatgccaccc tggaacacca 3060
gctgacacca tgggcgtcca cacctgctgc tccagacaag cacaaagcaa tctttcagcc 3120
ttgaaatgta ttatctgaaa ggctacctga agcccaggcc cgaatatggg gacttagtcg 3180
attacctgga aaaagaaaag acccacactg tgtcctgctg tgcttttggg caggaaaatg 3240
gaagaaagag tggggtgggc acattagaag tcacccaaat cctgccaggc tgcctggcat 3300
ccctggggca tgagctgggc ggagaatcca ccccgcagga tgttcagagg gacccactcc 3360
ttcatttttc agagtcaaag gaatcagagg ctcacccatg gcaggcagtg aaaagagcca 3420
ggagtcctgg gttctagtcc ctgctctgcc cccaactggc tgtataacct ttgaaaaatc 3480
attttctttg tctgagtctc tggttctccg tcagcaacag gctggcataa ggtcccctgc 3540
aggttccttc tagctggagc actcagagct tccctgactg ctagcagcct ctctggccct 3600
cacagggctg attgttctcc ttctccctgg agctctctct cctgaaaatc tccatcagag 3660
caaggcagcc agagaagccc ctgagaggga atgattggga agtgtccact ttctcaaccg 3720
gctcatcaaa cacactcctt tgtctatgaa tggcacatgt aaatgatgtt atattttgta 3780
tcttttatat catatgcttc accattctgt aaagggcctc tgcattgttg ctcccatcag 3840
gggtctcaag tggaaataaa ccctcgtgga taaccaaaaa aaaaaaaaaa aaaaa 3895
<210>107
<211>728
<212>PRT
<213>Homo sapiens
<220>
<223>mannan-binding lectin-associated serine protease-3
<400>107
Met Arg Trp Leu Leu Leu Tyr Tyr Ala Leu Cys Phe Ser Leu Ser Lys
1 5 10 15
Ala Ser Ala His Thr Val Glu Leu Asn Asn Met Phe Gly Gln Ile Gln
20 25 30
Ser Pro Gly Tyr Pro Asp Ser Tyr Pro Ser Asp Ser Glu Val Thr Trp
35 40 45
Asn Ile Thr Val Pro Asp Gly Phe Arg Ile Lys Leu Tyr Phe Met His
50 55 60
Phe Asn Leu Glu Ser Ser Tyr Leu Cys Glu Tyr Asp Tyr Val Lys Val
65 70 75 80
Glu Thr Glu Asp Gln Val Leu Ala Thr Phe Cys Gly Arg Glu Thr Thr
85 90 95
Asp Thr Glu Gln Thr Pro Gly Gln Glu Val Val Leu Ser Pro Gly Ser
100 105 110
Phe Met Ser Ile Thr Phe Arg Ser Asp Phe Ser Asn Glu Glu Arg Phe
115 120 125
Thr Gly Phe Asp Ala His Tyr Met Ala Val Asp Val Asp Glu Cys Lys
130 135 140
Glu Arg Glu Asp Glu Glu Leu Ser Cys Asp His Tyr Cys His Asn Tyr
145 150 155 160
Ile Gly Gly Tyr Tyr Cys Ser Cys Arg Phe Gly Tyr Ile Leu His Thr
165 170 175
Asp Asn Arg Thr Cys Arg Val Glu Cys Ser Asp Asn Leu Phe Thr Gln
180 185 190
Arg Thr Gly Val Ile Thr Ser Pro Asp Phe Pro Asn Pro Tyr Pro Lys
195 200 205
Ser Ser Glu Cys Leu Tyr Thr Ile Glu Leu Glu Glu Gly Phe Met Val
210 215 220
Asn Leu Gln Phe Glu Asp Ile Phe Asp Ile Glu Asp His Pro Glu Val
225 230 235 240
Pro Cys Pro Tyr Asp Tyr Ile Lys Ile Lys Val Gly Pro Lys Val Leu
245 250 255
Gly Pro Phe Cys Gly Glu Lys Ala Pro Glu Pro Ile Ser Thr Gln Ser
260 265 270
His Ser Val Leu Ile Leu Phe His Ser Asp Asn Ser Gly Glu Asn Arg
275 280 285
Gly Trp Arg Leu Ser Tyr Arg Ala Ala Gly Asn Glu Cys Pro Glu Leu
290 295 300
Gln Pro Pro Val His Gly Lys Ile Glu Pro Ser Gln Ala Lys Tyr Phe
305 310 315 320
Phe Lys Asp Gln Val Leu Val Ser Cys Asp Thr Gly Tyr Lys Val Leu
325 330 335
Lys Asp Asn Val Glu Met Asp Thr Phe Gln Ile Glu Cys Leu Lys Asp
340 345 350
Gly Thr Trp Ser Asn Lys Ile Pro Thr Cys Lys Ile Val Asp Cys Arg
355 360 365
Ala Pro Gly Glu Leu Glu His Gly Leu Ile Thr Phe Ser Thr Arg Asn
370 375 380
Asn Leu Thr Thr Tyr Lys Ser Glu Ile Lys Tyr Ser Cys Gln Glu Pro
385 390 395 400
Tyr Tyr Lys Met Leu Asn Asn Asn Thr Gly Ile Tyr Thr Cys Ser Ala
405 410 415
Gln Gly Val Trp Met Asn Lys Val Leu Gly Arg Ser Leu Pro Thr Cys
420 425 430
Leu Pro Glu Cys Gly Gln Pro Ser Arg Ser Leu Pro Ser Leu ValLys
435 440 445
Arg Ile Ile Gly Gly Arg Asn Ala Glu Pro Gly Leu Phe Pro Trp Gln
450 455 460
Ala Leu Ile Val Val Glu Asp Thr Ser Arg Val Pro Asn Asp Lys Trp
465 470 475 480
Phe Gly Ser Gly Ala Leu Leu Ser Ala Ser Trp Ile Leu Thr Ala Ala
485 490 495
His Val Leu Arg Ser Gln Arg Arg Asp Thr Thr Val Ile Pro Val Ser
500 505 510
Lys Glu His Val Thr Val Tyr Leu Gly Leu His Asp Val Arg Asp Lys
515 520 525
Ser Gly Ala Val Asn Ser Ser Ala Ala Arg Val Val Leu His Pro Asp
530 535 540
Phe Asn Ile Gln Asn Tyr Asn His Asp Ile Ala Leu Val Gln Leu Gln
545 550 555 560
Glu Pro Val Pro Leu Gly Pro His Val Met Pro Val Cys Leu Pro Arg
565 570 575
Leu Glu Pro Glu Gly Pro Ala Pro His Met Leu Gly Leu Val Ala Gly
580 585 590
Trp Gly Ile Ser Asn Pro Asn Val Thr Val Asp Glu Ile Ile Ser Ser
595 600 605
Gly Thr Arg Thr Leu Ser Asp Val Leu Gln Tyr Val Lys Leu Pro Val
610 615 620
Val Pro His Ala Glu Cys Lys Thr Ser Tyr Glu Ser Arg Ser Gly Asn
625 630 635 640
Tyr Ser Val Thr Glu Asn Met Phe Cys Ala Gly Tyr Tyr Glu Gly Gly
645 650 655
Lys Asp Thr Cys Leu Gly Asp Ser Gly Gly Ala Phe Val Ile Phe Asp
660 665 670
Asp Leu Ser Gln Arg Trp Val Val Gln Gly Leu Val Ser Trp Gly Gly
675 680 685
Pro Glu Glu Cys Gly Ser Lys Gln Val Tyr Gly Val Tyr Thr Lys Val
690 695 700
Ser Asn Tyr Val Asp Trp Val Trp Glu Gln Met Gly Leu Pro Gln Ser
705 710 715 720
Val Val Glu Pro Gln Val Glu Arg
725
<210>108
<211>924
<212>DNA
<213>Homo sapiens
<220>
<223>cathepsin G
<400>108
gcacagcagc aactgactgg gcagcctttc aggaaagatg cagccactcc tgcttctgct 60
ggcctttctc ctacccactg gggctgaggc aggggagatc atcggaggcc gggagagcag 120
gccccactcc cgcccctaca tggcgtatct tcagatccag agtccagcag gtcagagcag 180
atgtggaggg ttcctggtgc gagaagactt tgtgctgaca gcagctcatt gctggggaag 240
caatataaat gtcaccctgg gcgcccacaa tatccagaga cgggaaaaca cccagcaaca 300
catcactgcg cgcagagcca tccgccaccc tcaatataat cagcggacca tccagaatga 360
catcatgtta ttgcagctga gcagaagagt cagacggaat cgaaacgtga acccagtggc 420
tctgcctaga gcccaggagg gactgagacc cgggacgctg tgcactgtgg ccggctgggg 480
cagggtcagc atgaggaggg gaacagatac actccgagag gtgcagctga gagtgcagag 540
ggataggcag tgcctccgca tcttcggttc ctacgacccc cgaaggcaga tttgtgtggg 600
ggaccggcgg gaacggaagg ctgccttcaa gggggattcc ggaggccccc tgctgtgtaa 660
caatgtggcc cacggcatcg tctcctatgg aaagtcgtca ggggttcctc cagaagtctt 720
caccagggtc tcaagtttcc tgccctggat aaggacaaca atgagaagct tcaaactgct 780
ggatcagatg gagacccccc tgtgactgac tcttcttctc ggggacacag gccagctcca 840
cagtgttgcc agagccttaa taaacgtcca cagagtataa ataaccaatt cctcatttgt 900
tcattaaacg tcattcagta ctta 924
<210>109
<211>255
<212>PRT
<213>Homo sapiens
<220>
<223>cathepsin G
<400>109
Met Gln Pro Leu Leu Leu Leu Leu Ala Phe Leu Leu Pro Thr Gly Ala
1 5 10 15
Glu Ala Gly Glu Ile Ile Gly Gly Arg Glu Ser Arg Pro His Ser Arg
20 25 30
Pro Tyr Met Ala Tyr Leu Gln Ile Gln Ser Pro Ala Gly Gln Ser Arg
35 40 45
Cys Gly Gly Phe Leu Val Arg Glu Asp Phe Val Leu Thr Ala Ala His
50 55 60
Cys Trp Gly Ser Asn Ile Asn Val Thr Leu Gly Ala His Asn Ile Gln
65 70 75 80
Arg Arg Glu Asn Thr Gln Gln His Ile Thr Ala Arg Arg Ala Ile Arg
85 90 95
His Pro Gln Tyr Asn Gln Arg Thr Ile Gln Asn Asp Ile Met Leu Leu
100 105 110
Gln Leu Ser Arg Arg Val Arg Arg Asn Arg Asn Val Asn Pro ValAla
115 120 125
Leu Pro Arg Ala Gln Glu Gly Leu Arg Pro Gly Thr Leu Cys Thr Val
130 135 140
Ala Gly Trp Gly Arg Val Ser Met Arg Arg Gly Thr Asp Thr Leu Arg
145 150 155 160
Glu Val Gln Leu Arg Val Gln Arg Asp Arg Gln Cys Leu Arg Ile Phe
165 170 175
Gly Ser Tyr Asp Pro Arg Arg Gln Ile Cys Val Gly Asp Arg Arg Glu
180 185 190
Arg Lys Ala Ala Phe Lys Gly Asp Ser Gly Gly Pro Leu Leu Cys Asn
195 200 205
Asn Val Ala His Gly Ile Val Ser Tyr Gly Lys Ser Ser Gly Val Pro
210 215 220
Pro Glu Val Phe Thr Arg Val Ser Ser Phe Leu Pro Trp Ile Arg Thr
225 230 235 240
Thr Met Arg Ser Phe Lys Leu Leu Asp Gln Met Glu Thr Pro Leu
245 250 255
<210>110
<211>1001
<212>DNA
<213>Homo sapiens
<220>
<223>myeloblastin(proteinase 3)
<400>110
cgtgggtgca ccctggaccc caccatggct caccggcccc ccagccctgc cctggcgtcc 60
gtgctgctgg ccttgctgct gagcggtgct gcccgagctg cggagatcgt gggcgggcac 120
gaggcgcagc cacactcccg gccctacatg gcctccctgc agatgcgggg gaacccgggc 180
agccacttct gcggaggcac cttgatccac cccagcttcg tgctgacggc cgcgcactgc 240
ctgcgggaca taccccagcg cctggtgaac gtggtgctcg gagcccacaa cgtgcggacg 300
caggagccca cccagcagca cttctcggtg gctcaggtgt ttctgaacaa ctacgacgcg 360
gagaacaaac tgaacgacgt tctcctcatc cagctgagca gcccagccaa cctcagtgcc 420
tccgtcgcca cagtccagct gccacagcag gaccagccag tgccccacgg cacccagtgc 480
ctggccatgg gctggggccg cgtgggtgcc cacgaccccc cagcccaggt cctgcaggag 540
ctcaatgtca ccgtggtcac cttcttctgc cggccacata acatttgcac tttcgtccct 600
cgccgcaagg ccggcatctg cttcggagac tcaggtggcc ccctgatctg tgatggcatc 660
atccaaggaa tagactcctt cgtgatctgg ggatgtgcca cccgcctttt ccctgacttc 720
ttcacgcggg tagccctcta cgtggactgg atccgttcca cgctgcgccg tgtggaggcc 780
aagggccgcc cctgaaccgc ccctcccaca gcgctggccg ggaccccgag cctggctcca 840
aaccctcgag gcggatcttt ggacagaagc agctcttccc cgaacactgt ggcgtccggg 900
acggccccac ccgtcccccc acactccctc ccacggggct ccgggagaca ggccggccct 960
gcacctcacc ccaccgtgac ctcaataaac gttgaaactc c 1001
<210>111
<211>256
<212>PRT
<213>Homo sapiens
<220>
<223>myeloblastin (proteinase 3)
<400>111
Met Ala His Arg Pro Pro Ser Pro Ala Leu Ala Ser Val Leu Leu Ala
1 5 10 15
Leu Leu Leu Ser Gly Ala Ala Arg Ala Ala Glu Ile Val Gly Gly His
20 25 30
Glu Ala Gln Pro His Ser Arg Pro Tyr Met Ala Ser Leu Gln Met Arg
35 40 45
Gly Asn Pro Gly Ser His Phe Cys Gly Gly Thr Leu Ile His Pro Ser
50 55 60
Phe Val Leu Thr Ala Ala His Cys Leu Arg Asp Ile Pro Gln Arg Leu
65 70 75 80
Val Asn Val Val Leu Gly Ala His Asn Val Arg Thr Gln Glu Pro Thr
85 90 95
Gln Gln His Phe Ser Val Ala Gln Val Phe Leu Asn Asn Tyr Asp Ala
100 105 110
Glu Asn Lys Leu Asn Asp Val Leu Leu Ile Gln Leu Ser Ser Pro Ala
115 120 125
Asn Leu Ser Ala Ser Val Ala Thr Val Gln Leu Pro Gln Gln Asp Gln
130 135 140
Pro Val Pro His Gly Thr Gln Cys Leu Ala Met Gly Trp Gly Arg Val
145 150 155 160
Gly Ala His Asp Pro Pro Ala Gln Val Leu Gln Glu Leu Asn Val Thr
165 170 175
Val Val Thr Phe Phe Cys Arg Pro His Asn Ile Cys Thr Phe Val Pro
180 185 190
Arg Arg Lys Ala Gly Ile Cys Phe Gly Asp Ser Gly Gly Pro Leu Ile
195 200 205
Cys Asp Gly Ile Ile Gln Gly Ile Asp Ser Phe Val Ile Trp Gly Cys
210 215 220
Ala Thr Arg Leu Phe Pro Asp Phe Phe Thr Arg Val Ala Leu Tyr Val
225 230 235 240
Asp Trp Ile Arg Ser Thr Leu Arg Arg Val Glu Ala Lys Gly Arg Pro
245 250 255
<210>112
<211>878
<212>DNA
<213>Homo sapiens
<220>
<223>granzyme A
<400>112
cagattttca ggttgattga tgtgggacag cagccacaat gaggaactcc tatagatttc 60
tggcatcctc tctctcagtt gtcgtttctc tcctgctaat tcctgaagat gtctgtgaaa 120
aaattattgg aggaaatgaa gtaactcctc attcaagacc ctacatggtc ctacttagtc 180
ttgacagaaa aaccatctgt gctggggctt tgattgcaaa agactgggtg ttgactgcag 240
ctcactgtaa cttgaacaaa aggtcccagg tcattcttgg ggctcactca ataaccaggg 300
aagagccaac aaaacagata atgcttgtta agaaagagtt tccctatcca tgctatgacc 360
cagccacacg cgaaggtgac cttaaacttt tacagctgac ggaaaaagca aaaattaaca 420
aatatgtgac tatccttcat ctacctaaaa agggggatga tgtgaaacca ggaaccatgt 480
gccaagttgc agggtggggg aggactcaca atagtgcatc ttggtccgat actctgagag 540
aagtcaatat caccatcata gacagaaaag tctgcaatga tcgaaatcac tataatttta 600
accctgtgat tggaatgaat atggtttgtg ctggaagcct ccgaggtgga agagactcgt 660
gcaatggaga ttctggaagc cctttgttgt gcgagggtgt tttccgaggg gtcacttcct 720
ttggccttga aaataaatgc ggagaccctc gtgggcctgg tgtctatatt cttctctcaa 780
agaaacacct caactggata attatgacta tcaagggagc agtttaaata accgtttcct 840
ttcatttact gtggcttctt aatcttttca caaataaa 878
<210>113
<211>262
<212>PRT
<213>Homo sapiens
<220>
<223>granzyme A
<400>113
Met Arg Asn Ser Tyr Arg Phe Leu Ala Ser Ser Leu Ser Val Val Val
1 5 10 15
Ser Leu Leu Leu Ile Pro Glu Asp ValCys Glu Lys Ile Ile Gly Gly
20 25 30
Asn Glu Val Thr Pro His Ser Arg Pro Tyr Met Val Leu Leu Ser Leu
35 40 45
Asp Arg Lys Thr Ile Cys Ala Gly Ala Leu Ile Ala Lys Asp Trp Val
50 55 60
Leu Thr Ala Ala His Cys Asn Leu Asn Lys Arg Ser Gln Val Ile Leu
65 70 75 80
Gly Ala His Ser Ile Thr Arg Glu Glu Pro Thr Lys Gln Ile Met Leu
85 90 95
Val Lys Lys Glu Phe Pro Tyr Pro Cys Tyr Asp Pro Ala Thr Arg Glu
100 105 110
Gly Asp Leu Lys Leu Leu Gln Leu Thr Glu Lys Ala Lys Ile Asn Lys
115 120 125
Tyr Val Thr Ile Leu His Leu Pro Lys Lys Gly Asp Asp Val Lys Pro
130 135 140
Gly Thr Met Cys Gln Val Ala Gly Trp Gly Arg Thr His Asn Ser Ala
145 150 155 160
Ser Trp Ser Asp Thr Leu Arg Glu Val Asn Ile Thr Ile Ile Asp Arg
165 170 175
Lys Val Cys Asn Asp Arg Asn His Tyr Asn Phe Asn Pro Val Ile Gly
180 185 190
Met Asn Met Val Cys Ala Gly Ser Leu Arg Gly Gly Arg Asp Ser Cys
195 200 205
Ash Gly Asp Ser Gly Ser Pro Leu Leu Cys Glu Gly Val Phe Arg Gly
210 215 220
Val Thr Ser Phe Gly Leu Glu Asn Lys Cys Gly Asp Pro Arg Gly Pro
225 230 235 240
Gly Val Tyr Ile Leu Leu Ser Lys Lys His Leu Asn Trp Ile Ile Met
245 250 255
Thr Ile Lys Gly Ala Val
260
<210>114
<211>947
<212>DNA
<213>Homo sapiens
<220>
<223>granzyme M
<400>114
ggctcggggc cggggccagc acccacactg ggtctccaca gcggcatgga ggcctgcgtg 60
tcttcactgc tggtgctggc cctgggggcc ctgtcagtag gcagctcctt tgggacccag 120
atcatcgggg gccgggaggt gatcccccac tcgcgcccgt acatggcctc actgcagaga 180
aatggctccc acctgtgcgg gggtgtcctg gtgcacccaa agtgggtgct gacggctgcc 240
cactgcctgg cccagcggat ggcccagctg aggctggtgc tggggctcca caccctggac 300
agccccggtc tcaccttcca catcaaggca gccatccagc accctcgcta caagcccgtc 360
cctgccctgg agaacgacct cgcgctgctt cagctggacg ggaaagtgaa gcccagccgg 420
accatccggc cgttggccct gcccagtaag cgccaggtgg tggcagcagg gactcggtgc 480
agcatggccg gctgggggct gacccaccag ggcgggcgcc tgtcccgggt gctgcgggag 540
ctggacctcc aagtgctgga cacccgcatg tgtaacaaca gccgcttctg gaacggcagc 600
ctctccccca gcatggtctg cctggcggcc gactccaagg accaggctcc ctgcaagggt 660
gactcgggcg ggcccctggt gtgtggcaaa ggccgggtgt tggccggagt cctgtccttc 720
agctccaggg tctgcactga catcttcaag cctcccgtgg ccaccgctgt ggcgccttac 780
gtgtcctgga tcaggaaggt caccggccga tcggcctgat gccctggggt gatggggacc 840
ccctcgctgt ctccacagga cccttcccct ccaggggtgc agtggggtgg gtgaggacgg 900
gtgggaggga cagggaggga ccaataaatc ataatgaaga aacgctc 947
<210>115
<211>257
<212>PRT
<213>Homo sapiens
<220>
<223>granzyme M
<400>115
Met Glu Ala Cys Val Ser Ser Leu Leu Val Leu Ala Leu Gly Ala Leu
1 5 10 15
Ser Val Gly Ser Ser Phe Gly Thr Gln Ile Ile Gly Gly Arg Glu Val
20 25 30
Ile Pro His Ser Arg Pro Tyr Met Ala Ser Leu Gln Arg Asn Gly Ser
35 40 45
His Leu Cys Gly Gly Val Leu Val His Pro Lys Trp Val Leu Thr Ala
50 55 60
Ala His Cys Leu Ala Gln Arg Met Ala Gln Leu Arg Leu Val Leu Gly
65 70 75 80
Leu His Thr Leu Asp Ser Pro Gly Leu Thr Phe His Ile Lys Ala Ala
85 90 95
Ile Gln His Pro Arg Tyr Lys Pro Val Pro Ala Leu Glu Asn Asp Leu
100 105 110
Ala Leu Leu Gln Leu Asp Gly Lys Val Lys Pro Ser Arg Thr Ile Arg
115 120 125
Pro Leu Ala Leu Pro Ser Lys Arg Gln Val Val Ala Ala Gly Thr Arg
130 135 140
Cys Ser Met Ala Gly Trp Gly Leu Thr His Gln Gly Gly Arg Leu Ser
145 150 155 160
Arg Val Leu Arg Glu Leu Asp Leu Gln Val Leu Asp Thr Arg Met Cys
165 170 175
Asn Asn Ser Arg Phe Trp Asn Gly Ser Leu Ser Pro Ser Met Val Cys
180 185 190
Leu Ala Ala Asp Ser Lys Asp Gln Ala Pro Cys Lys Gly Asp Ser Gly
195 200 205
Gly Pro Leu Val Cys Gly Lys Gly Arg Val Leu Ala Gly Val Leu Ser
210 215 220
Phe Ser Ser Arg Val Cys Thr Asp Ile Phe Lys Pro Pro Val Ala Thr
225 230 235 240
Ala Val Ala Pro Tyr Val Ser Trp Ile Arg Lys Val Thr Gly Arg Ser
245 250 255
Ala
<210>116
<211>784
<212>DNA
<213>Homo sapiens
<220>
<223>chymase(human-type)
<400>116
agcatttgct caggcagcct ctctgggaag atgctgcttc ttcctctccc cctgctgctc 60
tttctcttgt gctccagagc tgaagctggg gagatcatcg ggggcacaga atgcaagcca 120
cattcccgcc cctacatggc ctacctggaa attgtaactt ccaacggtcc ctcaaaattt 180
tgtggtggtt tccttataag acggaacttt gtgctgacgg ctgctcattg tgcaggaagg 240
tctataacag tcacccttgg agcccataac ataacagagg aagaagacac atggcagaag 300
cttgaggtta taaagcaatt ccgtcatcca aaatataaca cttctactct tcaccacgat 360
atcatgttac taaagttgaa ggagaaagcc agcctgaccc tggctgtggg gacactcccc 420
ttcccatcac aattcaactt tgtcccacct gggagaatgt gccgggtggc tggctgggga 480
agaacaggtg tgttgaagcc gggctcagac actctgcaag aggtgaagct gagactcatg 540
gatccccagg cctgcagcca cttcagagac tttgaccaca atcttcagct gtgtgtgggc 600
aatcccagga agacaaaatc tgcatttaag ggagactctg ggggccctct tctgtgtgct 660
ggggtggccc agggcatcgt atcctatgga cggtcggatg caaagccccc tgctgtcttc 720
acccgaatct cccattaccg gccctggatc aaccagatcc tgcaggcaaa ttaatcctgg 780
atcc 784
<210>117
<211>247
<212>PRT
<213>Homo sapiens
<220>
<223>chymase(human-type)
<400>117
Met Leu Leu Leu Pro Leu Pro Leu Leu Leu Phe Leu Leu Cys Ser Arg
1 5 10 15
Ala Glu Ala Gly Glu Ile Ile Gly Gly Thr Glu Cys Lys Pro His Ser
20 25 30
Arg Pro Tyr Met Ala Tyr Leu Glu Ile Val Thr Ser Asn Gly Pro Ser
35 40 45
Lys Phe Cys Gly Gly Phe Leu Ile Arg Arg Asn Phe Val Leu Thr Ala
50 55 60
Ala His Cys Ala Gly Arg Ser Ile Thr Val Thr Leu Gly Ala His Asn
65 70 75 80
Ile Thr Glu Glu Glu Asp Thr Trp Gln Lys Leu Glu Val Ile Lys Gln
85 90 95
Phe Arg His Pro Lys Tyr Asn Thr Ser Thr Leu His His Asp Ile Met
100 105 110
Leu Leu Lys Leu Lys Glu Lys Ala Ser Leu Thr Leu Ala Val Gly Thr
115 120 125
Leu Pro Phe Pro Ser Gln Phe Asn Phe Val Pro Pro Gly Arg Met Cys
130 135 140
Arg Val Ala Gly Trp Gly Arg Thr Gly Val Leu Lys Pro Gly Ser Asp
145 150 155 160
Thr Leu Gln Glu Val Lys Leu Arg Leu Met Asp Pro Gln Ala Cys Ser
165 170 175
His Phe Arg Asp Phe Asp His Asn Leu Gln Leu Cys Val Gly Asn Pro
180 185 190
Arg Lys Thr Lys Ser Ala Phe Lys Gly Asp Ser Gly Gly Pro Leu Leu
195 200 205
Cys Ala Gly Val Ala Gln Gly Ile Val Ser Tyr Gly Arg Ser Asp Ala
210 215 220
Lys Pro Pro Ala Val Phe Thr Arg Ile Ser His Tyr Arg Pro Trp Ile
225 230 235 240
Asn Gln Ile Leu Gln Ala Asn
245
<210>118
<211>1194
<212>DNA
<213>Homo sapiens
<220>
<223>tryptase alpha (1)
<400>118
ataaatgggg aggggagagc ccactgggta gaaggaacag ggagcggcca ggatgctgaa 60
tctgctgctg ctggcgctgc ccgtcctggc gagccgcgcc tacgcggccc ctgccccagg 120
ccaggccctg cagcgagtgg gcatcgtcgg gggtcaggag gcccccagga gcaagtggcc 180
ctggcaggtg agcctgagag tccacggccc atactggatg cacttctgcg ggggctccct 240
catccacccc cagtgggtgc tgaccgcagc gcactgcgtg ggaccggacg tcaaggatct 300
ggccgccctc agggtgcaac tgcgggagca gcacctctac taccaggacc agctgctgcc 360
ggtcagcagg atcatcgtgc acccacagtt ctacaccgcc cagatcggag cggacatcgc 420
cctgctggag ctggaggagc cggtgaacgt ctccagccac gtccacacgg tcaccctgcc 480
ccctgcctca gagaccttcc ccccggggat gccgtgctgg gtcactggct ggggcgatgt 540
ggacaatgat gagcgcctcc caccgccatt tcctctgaag caggtgaagg tccccataat 600
ggaaaaccac atttgtgacg caaaatacca ccttggcgcc tacacgggag acgacgtccg 660
catcgtccgt gacgacatgc tgtgtgccgg gaacacccgg agggactcat gccagggcga 720
ctccggaggg cccctggtgt gcaaggtgaa tggcacctgg ctgcaggcgg gcgtggtcag 780
ctggggcgag ggctgtgccc agcccaaccg gcctggcatc tacacccgtg tcacctacta 840
cttggactgg atccaccact atgtccccaa aaagccgtga gtcaggcctg ggttggccac 900
ctgggtcact ggaggaccaa cccctgctgt ccaaaacacc actgcttcct acccaggtgg 960
cgactgcccc ccacaccttc cctgccccgt cctgagtgcc ccttcctgtc ctaagccccc 1020
tgctctcttc tgagcccctt cccctgtcct gaggaccctt ccctatcctg agcccccttc 1080
cctgtcctaa gcctgacgcc tgcaccgggc cctccagccc tcccctgccc agatagctgg 1140
tggtgggcgc taatcctcct gagtgctgga cctcattaaa gtgcatggaa atca 1194
<210>119
<211>275
<212>PRT
<213>Homo sapiens
<220>
<223>tryptase alpha (1)
<400>119
Met Leu Asn Leu Leu Leu Leu Ala Leu Pro Val Leu Ala Ser Arg Ala
1 5 10 15
Tyr Ala Ala Pro Ala Pro Gly Gln Ala Leu Gln Arg Val Gly Ile Val
20 25 30
Gly Gly Gln Glu Ala Pro Arg Ser Lys Trp Pro Trp Gln Val Ser Leu
35 40 45
Arg Val His Gly Pro Tyr Trp Met His Phe Cys Gly Gly Ser Leu Ile
50 55 60
His Pro Gln Trp Val Leu Thr Ala Ala His Cys Val Gly Pro Asp Val
65 70 75 80
Lys Asp Leu Ala Ala Leu Arg Val Gln Leu Arg Glu Gln His Leu Tyr
85 90 95
Tyr Gln Asp Gln Leu Leu Pro Val Ser Arg Ile Ile Val His Pro Gln
100 105 110
Phe Tyr Thr Ala Gln Ile Gly Ala Asp Ile Ala Leu Leu Glu Leu Glu
115 120 125
Glu Pro Val Asn Val Ser Ser His Val His Thr Val Thr Leu Pro Pro
130 135 140
Ala Ser Glu Thr Phe Pro Pro Gly Met Pro Cys Trp Val Thr Gly Trp
145 150 155 160
Gly Asp Val Asp Asn Asp Glu Arg Leu Pro Pro Pro Phe Pro Leu Lys
165 170 175
Gln ValLys Val Pro Ile Met Glu Asn His Ile Cys Asp Ala Lys Tyr
180 185 190
His Leu Gly Ala Tyr Thr Gly Asp Asp ValArg Ile Val Arg Asp Asp
195 200 205
Met Leu Cys Ala Gly Asn Thr Arg Arg Asp Ser Cys Gln Gly Asp Ser
210 215 220
Gly Gly Pro Leu Val Cys Lys Val Asn Gly Thr Trp Leu Gln Ala Gly
225 230 235 240
Val Val Ser Trp Gly Glu Gly Cys Ala Gln Pro Asn Arg Pro Gly Ile
245 250 255
Tyr Thr Arg Val Thr Tyr Tyr Leu Asp Trp Ile His His Tyr Val Pro
260 265 270
Lys Lys Pro
275
<210>120
<211>1074
<212>DNA
<213>Homo sapiens
<220>
<223>granzyme K
<400>120
gatcaacaca tttcatctgg gcttcttaaa tctaaatctt taaaatgact aagttttctt 60
ccttttctct gtttttccta atagttgggg cttatatgac tcatgtgtgt ttcaatatgg 120
aaattattgg agggaaagaa gtgtcacctc attccaggcc atttatggcc tccatccagt 180
atggcggaca tcacgtttgt ggaggtgttc tgattgatcc acagtgggtg ctgacagcag 240
cccactgcca atatcggttt accaaaggcc agtctcccac tgtggtttta ggcgcacact 300
ctctctcaaa gaatgaggcc tccaaacaaa cactggagat caaaaaattt ataccattct 360
caagagttac atcagatcct caatcaaatg atatcatgct ggttaagctt caaacagccg 420
caaaactcaa taaacatgtc aagatgctcc acataagatc caaaacctct cttagatctg 480
gaaccaaatg caaggttact ggctggggag ccaccgatcc agattcatta agaccttctg 540
acaccctgcg agaagtcact gttactgtcc taagtcgaaa actttgcaac agccaaagtt 600
actacaacgg cgaccctttt atcaccaaag acatggtctg tgcaggagat gccaaaggcc 660
agaaggattc ctgtaagggt gactcagggg gccccttgat ctgtaaaggt gtcttccacg 720
ctatagtctc tggaggtcat gaatgtggtg ttgccacaaa gcctggaatc tacaccctgt 780
taaccaagaa ataccagact tggatcaaaa gcaaccttgt cccgcctcat acaaattaag 840
ttacaaataa ttttattgga tgcacttgct tcttttttcc taatatgctc gcaggttaga 900
gttgggtgta agtaaagcag agcacatatg gggtccattt ttgcacttgt aagtcatttt 960
attaaggaat caagttcttt ttcacttgta tcactgatgt atttctacca tgctggtttt 1020
attctaaata aaatttagaa gactcaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 1074
<210>121
<211>264
<212>PRT
<213>Homo sapiens
<220>
<223>granzyme K
<400>121
Met Thr Lys Phe Ser Ser Phe Ser Leu Phe Phe Leu Ile Val Gly Ala
1 5 10 15
Tyr Met Thr His Val Cys Phe Asn Met Glu Ile Ile Gly Gly Lys Glu
20 25 30
Val Ser Pro His Ser Arg Pro Phe Met Ala Ser Ile Gln Tyr Gly Gly
35 40 45
His His Val Cys Gly Gly Val Leu Ile Asp Pro Gln Trp Val Leu Thr
50 55 60
Ala Ala His Cys Gln Tyr Arg Phe Thr Lys Gly Gln Ser Pro Thr Val
65 70 75 80
Val Leu Gly Ala His Ser Leu Ser Lys Asn Glu Ala Ser Lys Gln Thr
85 90 95
Leu Glu Ile Lys Lys Phe Ile Pro Phe Ser Arg Val Thr Ser Asp Pro
100 105 110
Gln Ser Asn Asp Ile Met Leu Val Lys Leu Gln Thr Ala Ala Lys Leu
115 120 125
Asn Lys His Val Lys Met Leu His Ile Arg Ser Lys Thr Ser Leu Arg
130 135 140
Ser Gly Thr Lys Cys Lys Val Thr Gly Trp Gly Ala Thr Asp Pro Asp
145 150 155 160
Ser Leu Arg Pro Ser Asp Thr Leu Arg Glu Val Thr Val Thr Val Leu
165 170 175
Ser Arg Lys Leu Cys Asn Ser Gln Ser Tyr Tyr Asn Gly Asp Pro Phe
180 185 190
Ile Thr Lys Asp Met ValCys Ala Gly Asp Ala Lys Gly Gln Lys Asp
195 200 205
Ser Cys Lys Gly Asp Ser Gly Gly Pro Leu Ile Cys Lys Gly Val Phe
210 215 220
His Ala Ile Val Ser Gly Gly His Glu Cys Gly Val Ala Thr Lys Pro
225 230 235 240
Gly Ile Tyr Thr Leu Leu Thr Lys Lys Tyr Gln Thr Trp Ile Lys Ser
245 250 255
Asn Leu Val Pro Pro His Thr Asn
260
<210>122
<211>1047
<212>DNA
<213>Homo sapiens
<220>
<223>granzyme H
<400>122
ggagtcaaca ccaacagctc tgacctgggc agccttcctg agaaaatgca gccattcctc 60
ctcctgttgg cctttcttct gacccctggg gctgggacag aggagatcat cgggggccat 120
gaggccaagc cccactcccg cccctacatg gcctttgttc agtttctgca agagaagagt 180
cggaagaggt gtggcggcat cctagtgaga aaggactttg tgctgacagc tgctcactgc 240
cagggaagct ccataaatgt caccttgggg gcccacaata tcaaggaaca ggagcggacc 300
cagcagttta tccctgtgaa aagacccatc ccccatccag cctataatcc taagaacttc 360
tccaacgaca tcatgctact gcagctggag agaaaggcca agtggaccac agctgtgcgg 420
cctctcaggc tacctagcag caaggcccag gtgaagccag ggcagctgtg cagtgtggct 480
ggctggggtt atgtctcaat gagcacttta gcaaccacac tgcaggaagt gttgctgaca 540
gtgcagaagg actgccagtg tgaacgtctc ttccatggca attacagcag agccactgag 600
atttgtgtgg gggatccaaa gaagacacag accggtttca agggggactc cggggggccc 660
ctcgtgtgta aggacgtagc ccaaggtatt ctctcctatg gaaacaaaaa agggacacct 720
ccaggagtct acatcaaggt ctcacacttc ctgccctgga taaagagaac aatgaagcgc 780
ctctaacagc aggcatgaga ctaaccttcc tctgggcctg accatctctg ggacagaggc 840
aagaatcccc aaggggtggg cagtcagggt tgcaggactg taataaatgg atctctggtg 900
tagaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 960
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1020
aaaaaaaaaa aaaaaaaaaa aaaaaaa 1047
<210>123
<211>246
<212>PRT
<213>Homo sapiens
<220>
<223>granzyme H
<400>123
Met Gln Pro Phe Leu Leu Leu Leu Ala Phe Leu Leu Thr Pro Gly Ala
1 5 10 15
Gly Thr Glu Glu Ile Ile Gly Gly His Glu Ala Lys Pro His Ser Arg
20 25 30
Pro Tyr Met Ala Phe Val Gln Phe Leu Gln Glu Lys Ser Arg Lys Arg
35 40 45
Cys Gly Gly Ile Leu Val Arg Lys Asp Phe Val Leu Thr Ala Ala His
50 55 60
Cys Gln Gly Ser Ser Ile Asn Val Thr Leu Gly Ala His Asn Ile Lys
65 70 75 80
Glu Gln Glu Arg Thr Gln Gln Phe Ile Pro Val Lys Arg Pro Ile Pro
85 90 95
His Pro Ala Tyr Asn Pro Lys Asn Phe Ser Asn Asp Ile Met Leu Leu
100 105 110
Gln Leu Glu Arg Lys Ala Lys Trp Thr Thr Ala Val Arg Pro Leu Arg
115 120 125
Leu Pro Ser Ser Lys Ala Gln Val Lys Pro Gly Gln Leu Cys Ser Val
130 135 140
Ala Gly Trp Gly Tyr Val Ser Met Ser Thr Leu Ala Thr Thr Leu Gln
145 150 155 160
Glu Val Leu Leu Thr Val Gln Lys Asp Cys Gln Cys Glu Arg Leu Phe
165 170 175
His Gly Asn Tyr Ser Arg Ala Thr Glu Ile Cys Val Gly Asp Pro Lys
180 185 190
Lys Thr Gln Thr Gly Phe Lys Gly Asp Ser Gly Gly Pro Leu Val Cys
195 200 205
Lys Asp Val Ala Gln Gly Ile Leu Ser Tyr Gly Asn Lys Lys Gly Thr
210 215 220
Pro Pro Gly Val Tyr Ile Lys Val Ser His Phe Leu Pro Trp Ile Lys
225 230 235 240
Arg Thr Met Lys Arg Leu
245
<210>124
<211>865
<212>DNA
<213>Homo sapiens
<220>
<223>chymotrypsin B
<400>124
ccctgccagc ggcaccatgg ctttcctctg gctcctctcc tgctgggccc tcctgggtac 60
caccttcggc tgcggggtcc ccgccatcca ccctgtgctc agcggcctgt ccaggatcgt 120
gaatggggag gacgccgtcc ccggctcctg gccctggcag gtgtccctgc aggacaaaac 180
cggcttccac ttctgcgggg gctccctcat cagcgaggac tgggtggtca ccgctgccca 240
ctgcggggtc aggacctccg acgtggtcgt ggctggggag tttgaccagg gctctgacga 300
ggagaacatc caggtcctga agatcgccaa ggtcttcaag aaccccaagt tcagcattct 360
gaccgtgaac aatgacatca ccctgctgaa gctggccaca cctgcccgct tctcccagac 420
agtgtccgcc gtgtgcctgc ccagcgccga cgacgacttc cccgcgggga cactgtgtgc 480
caccacaggc tggggcaaga ccaagtacaa cgccaacaag acccctgaca agctgcagca 540
ggcagccctg cccctcctgt ccaatgccga atgcaagaag tcctggggca ggaggatcac 600
cgacgtgatg atctgtgccg gggccagtgg cgtctcctcc tgcatgggcg actctggcgg 660
tcccctggtc tgccaaaagg atggagcctg gaccctggtg ggcattgtgt cctggggcag 720
cgacacctgc tccacctcca gccctggcgt gtacgcccgt gtcaccaagc tcataccttg 780
ggtgcagaag atcctggctg ccaactgagc ccgcggctcc ctccgaccct gctccccaca 840
gagcctcagt aaacccatgg aacac 865
<210>125
<211>263
<212>PRT
<213>Homo sapiens
<220>
<223>chymotrypsin B
<400>125
Met Ala Phe Leu Trp Leu Leu Ser Cys Trp Ala Leu Leu Gly Thr Thr
1 5 10 15
Phe Gly Cys Gly Val Pro Ala Ile His Pro Val Leu Ser Gly Leu Ser
20 25 30
Arg Ile Val Asn Gly Glu Asp Ala Val Pro Gly Ser Trp Pro Trp Gln
35 40 45
Val Ser Leu Gln Asp Lys Thr Gly Phe His Phe Cys Gly Gly Ser Leu
50 55 60
Ile Ser Glu Asp Trp Val Val Thr Ala Ala His Cys Gly Val Arg Thr
65 70 75 80
Ser Asp Val Val Val Ala Gly Glu Phe Asp Gln Gly Ser Asp Glu Glu
85 90 95
Asn Ile Gln Val Leu Lys Ile Ala Lys Val Phe Lys Asn Pro Lys Phe
100 105 110
Ser Ile Leu Thr Val Asn Asn Asp Ile Thr Leu Leu Lys Leu Ala Thr
115 120 125
Pro Ala Arg Phe Ser Gln Thr Val Ser Ala Val Cys Leu Pro Ser Ala
130 135 140
Asp Asp Asp Phe Pro Ala Gly Thr Leu Cys Ala Thr Thr Gly Trp Gly
145 150 155 160
Lys Thr Lys Tyr Asn Ala Asn Lys Thr Pro Asp Lys Leu Gln Gln Ala
165 170 175
Ala Leu Pro Leu Leu Ser Asn Ala Glu Cys Lys Lys Ser Trp Gly Arg
180 185 190
Arg Ile Thr Asp Val Met Ile Cys Ala Gly Ala Ser Gly ValSer Ser
195 200 205
Cys Met Gly Asp Ser Gly Gly Pro Leu Val Cys Gln Lys Asp Gly Ala
210 215 220
Trp Thr Leu Val Gly Ile Val Ser Trp Gly Ser Asp Thr Cys Ser Thr
225 230 235 240
Ser Ser Pro Gly Val Tyr Ala Arg Val Thr Lys Leu Ile Pro Trp Val
245 250 255
Gln Lys Ile Leu Ala Ala Asn
260
<210>126
<211>952
<212>DNA
<213>Homo sapiens
<220>
<223>pancreatic elastase
<400>126
ttggtccaag caagaaggca gtggtctact ccatcggcaa catgctggtc ctttatggac 60
acagcaccca ggaccttccg gaaaccaatg cccgcgtagt cggagggact gaggccggga 120
ggaattcctg gccctctcag atttccctcc agtaccggtc tggaggttcc cggtatcaca 180
cctgtggagg gacccttatc agacagaact gggtgatgac agctgctcac tgcgtggatt 240
accagaagac tttccgcgtg gtggctggag accataacct gagccagaat gatggcactg 300
agcagtacgt gagtgtgcag aagatcgtgg tgcatccata ctggaacagc gataacgtgg 360
ctgccggcta tgacatcgcc ctgctgcgcc tggcccagag cgttaccctc aatagctatg 420
tccagctggg tgttctgccc caggagggag ccatcctggc taacaacagt ccctgctaca 480
tcacaggctg gggcaagacc aagaccaatg ggcagctggc ccagaccctg cagcaggctt 540
acctgccctc tgtggactac gccatctgct ccagctcctc ctactggggc tccactgtga 600
agaacaccat ggtgtgtgct ggtggagatg gagttcgctc tggatgccag ggtgactctg 660
ggggccccct ccattgcttg gtgaatggca agtattctgt ccatggagtg accagctttg 720
tgtccagccg gggctgtaat gtctccagga agcctacagt cttcacccag gtctctgctt 780
acatctcctg gataaataat gtcatcgcct ccaactgaac attttcctga gtccaacgac 840
cttcccaaaa tggttcttag atctgcaata ggacttgcga tcaaaaagta aaacacattc 900
tgaaagacta ttgagccatt gatagaaaag caaataaaac tagatataca tt 952
<210>127
<211>258
<212>PRT
<213>Homo sapiens
<220>
<223>pancreatic elastase
<400>127
Met Leu Val Leu Tyr Gly His Ser Thr Gln Asp Leu Pro Glu Thr Asn
1 5 10 15
Ala Arg Val Val Gly Gly Thr Glu Ala Gly Arg Asn Ser Trp Pro Ser
20 25 30
Gln Ile Ser Leu Gln Tyr Arg Ser Gly Gly Ser Arg Tyr His Thr Cys
35 40 45
Gly Gly Thr Leu Ile Arg Gln Asn Trp Val Met Thr Ala Ala His Cys
50 55 60
Val Asp Tyr Gln Lys Thr Phe Arg Val Val Ala Gly Asp His Asn Leu
65 70 75 80
Ser Gln Asn Asp Gly Thr Glu Gln Tyr Val Ser Val Gln Lys Ile Val
85 90 95
Val His Pro Tyr Trp Asn Ser Asp Asn Val Ala Ala Gly Tyr Asp Ile
100 105 110
Ala Leu Leu Arg Leu Ala Gln Ser Val Thr Leu Asn Ser Tyr Val Gln
115 120 125
Leu Gly Val Leu Pro Gln Glu Gly Ala Ile Leu Ala Asn Asn Ser Pro
130 135 140
Cys Tyr Ile Thr Gly Trp Gly Lys Thr Lys Thr Asn Gly Gln Leu Ala
145 150 155 160
Gln Thr Leu Gln Gln Ala Tyr Leu Pro Ser Val Asp Tyr Ala Ile Cys
165 170 175
Ser Ser Ser Ser Tyr Trp Gly Ser Thr Val Lys Asn Thr Met Val Cys
180 185 190
Ala Gly Gly Asp Gly Val Arg Ser Gly Cys Gln Gly Asp Ser Gly Gly
195 200 205
Pro Leu His Cys Leu Val Asn Gly Lys Tyr Ser Val His Gly Val Thr
210 215 220
Ser Phe Val Ser Ser Arg Gly Cys Asn Val Ser Arg Lys Pro Thr Val
225 230 235 240
Phe Thr Gln Val Ser Ala Tyr Ile Ser Trp Ile Asn Asn Val Ile Ala
245 250 255
Ser Asn
<210>128
<211>963
<212>DNA
<213>Homo sapiens
<220>
<223>pancreatic endopeptidase E (A)
<400>128
tcctatcatc acaaaactca tgatgctccg gctgctcagt tccctcctcc ttgtggccgt 60
tgcctcaggc tatggcccac cttcctctca ctcttccagc cgcgttgtcc atggtgagga 120
tgcggtcccc tacagctggc cctggcaggt ttccctgcag tatgagaaaa gtggaagctt 180
ctaccacacg tgtggcggta gcctcatcgc ccccgattgg gttgtgactg ccggccactg 240
catctcgagg gatctgacct accaggtggt gttgggtgag tacaaccttg ctgtgaagga 300
gggccccgag caggtgatcc ccatcaactc tgaggagctg tttgtgcatc cactctggaa 360
ccgctcgtgt gtggcctgtg gcaatgacat cgccctcatc aagctctcac gcagcgccca 420
gctgggagat gccgtccagc tcgcctcact ccctcccgct ggtgacatcc ttcccaacaa 480
gacaccctgc tacatcaccg gctggggccg tctctatacc aatgggccac tcccagacaa 540
gctgcagcag gcccggctgc ccgtggtgga ctataagcac tgctccaggt ggaactggtg 600
gggttccacc gtgaagaaaa ccatggtgtg tgctggaggg tacatccgct ccggctgcaa 660
cggtgactct ggaggacccc tcaactgccc cacagaggat ggtggctggc aggtccacgg 720
tgtgaccagc tttgtttctg gctttggctg caacttcatc tggaagccca cggtgttcac 780
tcgagtctcc gccttcatcg actggattga ggagaccata gcaagccact agaaccaagg 840
cccagctggc agtgctgatc gatcccacat cctgaataaa gaataaagat ctctcagaaa 900
attccaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 960
aaa 963
<210>129
<211>270
<212>PRT
<213>Homo sapiens
<220>
<223>pancreatic endopeptidase E (A)
<400>129
Met Met Leu Arg Leu Leu Ser Ser Leu Leu Leu Val Ala Val Ala Ser
1 5 10 15
Gly Tyr Gly Pro Pro Ser Ser His Ser Ser Ser Arg Val Val His Gly
20 25 30
Glu Asp Ala Val Pro Tyr Ser Trp Pro Trp Gln Val Ser Leu Gln Tyr
35 40 45
Glu Lys Ser Gly Ser Phe Tyr His Thr Cys Gly Gly Ser Leu Ile Ala
50 55 60
Pro Asp Trp Val Val Thr Ala Gly His Cys Ile Ser Arg Asp Leu Thr
65 70 75 80
Tyr Gln Val Val Leu Gly Glu Tyr Asn Leu Ala Val Lys Glu Gly Pro
85 90 95
Glu Gln Val Ile Pro Ile Asn Ser Glu Glu Leu Phe Val His Pro Leu
100 105 110
Trp Asn Arg Ser Cys Val Ala Cys Gly Asn Asp Ile Ala Leu Ile Lys
115 120 125
Leu Ser Arg Ser Ala Gln Leu Gly Asp Ala Val Gln Leu Ala Ser Leu
130 135 140
Pro Pro Ala Gly Asp Ile Leu Pro Asn Lys Thr Pro Cys Tyr Ile Thr
145 150 155 160
Gly Trp Gly Arg Leu Tyr Thr Asn Gly Pro Leu Pro Asp Lys Leu Gln
165 170 175
Gln Ala Arg Leu Pro Val Val Asp Tyr Lys His Cys Ser Arg Trp Asn
180 185 190
Trp Trp Gly Ser Thr Val Lys Lys Thr Met Val Cys Ala Gly Gly Tyr
195 200 205
Ile Arg Ser Gly Cys Asn Gly Asp Ser Gly Gly Pro Leu Asn Cys Pro
210 215 220
Thr Glu Asp Gly Gly Trp Gln Val His Gly Val Thr Ser Phe Val Ser
225 230 235 240
Gly Phe Gly Cys Asn Phe Ile Trp Lys Pro Thr Val Phe Thr Arg Val
245 250 255
Ser Ala Phe Ile Asp Trp Ile Glu Glu Thr Ile Ala Ser His
260 265 270
<210>130
<211>914
<212>DNA
<213>Homo sapiens
<220>
<223>pancreatic elastase II (IIA)
<400>130
ttttacagaa ctcccacgga cacaccatga taaggacgct gctgctgtcc actttggtgg 60
ctggagccct cagttgtggg gaccccactt acccacctta tgtgactagg gtggttggcg 120
gtgaagaagc gaggcccaac agctggccct ggcaggtctc cctgcagtac agctccaatg 180
gcaagtggta ccacacctgc ggagggtccc tgatagccaa cagctgggtc ctgacggctg 240
cccactgcat cagctcctcc aggacctacc gcgtggggct gggccggcac aacctctacg 300
ttgcggagtc cggctcgctg gcagtcagtg tctctaagat tgtggtgcac aaggactgga 360
actccaacca aatctccaaa gggaacgaca ttgccctgct caaactggct aaccccgtct 420
ccctcaccga caagatccag ctggcctgcc tccctcctgc cggcaccatt ctacccaaca 480
actacccctg ctacgtcacg ggctggggaa ggctgcagac caacggggct gttcctgatg 540
tcctgcagca gggccggttg ctggttgtgg actatgccac ctgctccagc tctgcctggt 600
ggggcagcag cgtgaaaacc agtatgatct gtgctggggg tgatggcgtg atctccagct 660
gcaacggaga ctctggcggg ccactgaact gtcaggcgtc tgacggccgg tggcaggtgc 720
acggcatcgt cagcttcggg tctcgcctcg gctgcaacta ctaccacaag ccctccgtct 780
tcacgcgggt ctccaattac atcgactgga tcaattcggt gattgcaaat aactaaccaa 840
aagaagtccc tgggactgtt tcagacttgg aaaggtcaca gaaggaaaat aatataataa 900
agtgacaact atgc 914
<210>131
<211>269
<212>PRT
<213>Homo sapiens
<220>
<223>pancreatic elastase II (IIA)
<400>131
Met Ile Arg Thr Leu Leu Leu Ser Thr Leu Val Ala Gly Ala Leu Ser
1 5 10 15
Cys Gly Asp Pro Thr Tyr Pro Pro Tyr Val Thr Arg Val Val Gly Gly
20 25 30
Glu Glu Ala Arg Pro Asn Ser Trp Pro Trp Gln Val Ser Leu Gln Tyr
35 40 45
Ser Ser Asn Gly Lys Trp Tyr His Thr Cys Gly Gly Ser Leu Ile Ala
50 55 60
Asn Ser Trp Val Leu Thr Ala Ala His Cys Ile Ser Ser Ser Arg Thr
65 70 75 80
Tyr Arg Val Gly Leu Gly Arg His Asn Leu Tyr Val Ala Glu Ser Gly
85 90 95
Ser Leu Ala Val Ser Val Ser Lys Ile Val Val His Lys Asp Trp Asn
100 105 110
Ser Asn Gln Ile Ser Lys Gly Asn Asp Ile Ala Leu Leu Lys Leu Ala
115 120 125
Asn Pro Val Ser Leu Thr Asp Lys Ile Gln Leu Ala Cys Leu Pro Pro
130 135 140
Ala Gly Thr Ile Leu Pro Asn Asn Tyr Pro Cys Tyr Val Thr Gly Trp
145 150 155 160
Gly Arg Leu Gln Thr Asn Gly Ala Val Pro Asp Val Leu Gln Gln Gly
165 170 175
Arg Leu Leu Val Val Asp Tyr Ala Thr Cys Ser Ser Ser Ala Trp Trp
180 185 190
Gly Ser Ser Val Lys Thr Ser Met Ile Cys Ala Gly Gly Asp Gly Val
195 200 205
Ile Ser Ser Cys Asn Gly Asp Ser Gly Gly Pro Leu Asn Cys Gln Ala
210 215 220
Ser Asp Gly Arg Trp Gln Val His Gly Ile Val Ser Phe Gly Ser Arg
225 230 235 240
Leu Gly Cys Asn Tyr Tyr His Lys Pro Ser Val Phe Thr Arg Val Ser
245 250 255
Asn Tyr Ile Asp Trp Ile Asn Ser Val Ile Ala Asn Asn
260 265
<210>132
<211>3696
<212>DNA
<213>Homo sapiens
<220>
<223>Enteropeptidase
<400>132
accagacagt tcttaaatta gcaagccttc aaaaccaaaa atggggtcga aaagaggcat 60
atcttctagg catcattctc tcagctccta tgaaatcatg tttgcagctc tctttgccat 120
attggtagtg ctctgtgctg gattaattgc agtatcctgc ctgacaatca aggaatccca 180
acgaggtgca gcacttggac agagtcatga agccagagcg acatttaaaa taacatccgg 240
agttacatat aatcctaatt tgcaagacaa actctcagtg gatttcaaag ttcttgcttt 300
tgaccttcag caaatgatag atgagatctt tctatcaagc aatctgaaga atgaatataa 360
gaactcaaga gttttacaat ttgaaaatgg cagcattata gtcgtatttg accttttctt 420
tgcccagtgg gtgtcagatc aaaatgtaaa agaagaactg attcaaggcc ttgaagcaaa 480
taaatccagc caactggtca ctttccatat tgatttgaac agcgttgata tcctagacaa 540
gctaacaacc accagtcatc tggcaactcc aggaaatgtc tcaatagagt gcctgcctgg 600
ttcaagtcct tgtactgatg ctctaacgtg tataaaagct gatttatttt gtgatggaga 660
agtaaactgt ccagatggtt ctgacgaaga caataaaatg tgtgccacag tttgtgatgg 720
aagatttttg ttaactggat catctgggtc tttccaggct actcattatc caaaaccttc 780
tgaaacaagt gttgtctgcc agtggatcat acgtgtaaac caaggacttt ccattaaact 840
gagcttcgat gattttaata catattatac agatatatta gatatttatg aaggtgtagg 900
atcaagcaag attttaagag cttctatttg ggaaactaat cctggcacaa taagaatttt 960
ttccaaccaa gttactgcca cctttcttat agaatctgat gaaagtgatt atgttggctt 1020
taatgcaaca tatactgcat ttaacagcag tgagcttaat aattatgaga aaattaattg 1080
taactttgag gatggctttt gtttctgggt ccaggatcta aatgatgata atgaatggga 1140
aaggattcag ggaagcacct tttctccttt tactggaccc aattttgacc acacttttgg 1200
caatgcttca ggattttaca tttctacccc aactggacca ggagggagac aagaacgagt 1260
ggggctttta agcctccctt tggaccccac tttggagcca gcttgcctta gtttctggta 1320
tcatatgtat ggtgaaaatg tccataaatt aagcattaat atcagcaatg accaaaatat 1380
ggagaagaca gttttccaaa aggaaggaaa ttatggagac aattggaatt atggacaagt 1440
aaccctaaat gaaacagtta aatttaaggt tgcttttaat gcttttaaaa acaagatcct 1500
gagtgatatt gcgttggatg acattagcct aacatatggg atttgcaatg ggagtcttta 1560
tccagaacca actttggtgc caactcctcc accagaactt cctacggact gtggaggacc 1620
ttttgagctg tgggagccaa atacaacatt cagttctacg aactttccaa acagctaccc 1680
taatctggct ttctgtgttt ggattttaaa tgcacaaaaa ggaaagaata tacaacttca 1740
ttttcaagaa tttgacttag aaaatattaa cgatgtagtt gaaataagag atggtgaaga 1800
agctgattcc ttgctcttag ctgtgtacac agggcctggc ccagtaaagg atgtgttctc 1860
taccaccaac agaatgactg tgcttctcat cactaacgat gtgttggcaa gaggagggtt 1920
taaagcaaac tttactactg gctatcactt ggggattcca gagccatgca aggcagacca 1980
ttttcaatgt aaaaatggag agtgtgttcc actggtgaat ctctgtgacg gtcatctgca 2040
ctgtgaggat ggctcagatg aagcagattg tgtgcgtttt ttcaatggca caacgaacaa 2100
caatggttta gtgcggttca gaatccagag catatggcat acagcttgtg ctgagaactg 2160
gaccacccag atttcaaatg atgtttgtca actgctggga ctagggagtg gaaactcatc 2220
aaagccaatc ttctctaccg atggtggacc atttgtcaaa ttaaacacag cacctgatgg 2280
ccacttaata ctaacaccca gtcaacagtg tttacaggat tccttgattc ggttacagtg 2340
taaccataaa tcttgtggaa aaaaactggc agctcaagac atcaccccaa agattgttgg 2400
aggaagtaat gccaaagaag gggcctggcc ctgggttgtg ggtctgtatt atggcggccg 2460
actgctctgc ggcgcatctc tcgtcagcag tgactggctg gtgtccgccg cacactgcgt 2520
gtatgggaga aacttagagc catccaagtg gacagcaatc ctaggcctgc atatgaaatc 2580
aaatctgacc tctcctcaaa cagtccctcg attaatagat gaaattgtca taaaccctca 2640
ttacaatagg cgaagaaagg acaacgacat tgccatgatg catctggaat ttaaagtgaa 2700
ttacacagat tacatacaac ctatttgttt accggaagaa aatcaagttt ttcctccagg 2760
aagaaattgt tctattgctg gttgggggac ggttgtatat caaggtacta ctgcaaacat 2820
attgcaagaa gctgatgttc ctcttctatc aaatgagaga tgccaacagc agatgccaga 2880
atataacatt actgaaaata tgatatgtgc aggctatgaa gaaggaggaa tagattcttg 2940
tcagggggat tcaggaggac cattaatgtg ccaagaaaac aacaggtggt tccttgctgg 3000
tgtgacctca tttggataca agtgtgccct gcctaatcgc cccggagtgt atgccagggt 3060
ctcaaggttt accgaatgga tacaaagttt tctacattag cgcatttctt aaactaaaca 3120
ggaaagtcgc attattttcc cattctactc tagaaagcat ggaaattaag tgtttcgtac 3180
aaaaatttta aaaagttacc aaaggttttt attcttacct atgtcaatga aatgctaggg 3240
ggccagggaa acaaaatttt aaaaataata aaattcacca tagcaataca gaataacttt 3300
aaaataccat taaatacatt tgtatttcat tgtgaacagg tatttcttca cagatctcat 3360
ttttaaaatt cttaatgatt atttttatta cttactgttg tttaaaggga tgttatttta 3420
aagcatatac catacactta agaaatttga gcagaattta aaaaagaaag aaaataaatt 3480
gtttttccca aagtatgtca ctgttggaaa taaactgcca taaattttct agttccagtt 3540
tagtttgctg ctattagcag aaactcaatt gtttctctgt cttttctatc aaaattttca 3600
acatatgcat aaccttagta ttttcccaac caatagaaac tatttattgt aagcttatgt 3660
cacaggcctg gactaaattg attttacgtt cctctt 3696
<210>133
<211>1019
<212>PRT
<213>Homo sapiens
<220>
<223>Enteropeptidase
<400>133
Met Gly Ser Lys Arg Gly Ile Ser Ser Arg His His Ser Leu Ser Ser
1 5 10 15
Tyr Glu Ile Met Phe Ala Ala Leu Phe Ala Ile Leu Val Val Leu Cys
20 25 30
Ala Gly Leu Ile Ala Val Ser Cys Leu Thr Ile Lys Glu Ser Gln Arg
35 40 45
Gly Ala Ala Leu Gly Gln Ser His Glu Ala Arg Ala Thr Phe Lys Ile
50 55 60
Thr Ser Gly Val Thr Tyr Asn Pro Asn Leu Gln Asp Lys Leu Ser Val
65 70 75 80
Asp Phe Lys Val Leu Ala Phe Asp Leu Gln Gln Met Ile Asp Glu Ile
85 90 95
Phe Leu Ser Ser Asn Leu Lys Asn Glu Tyr Lys Asn Ser Arg Val Leu
100 105 110
Gln Phe Glu Asn Gly Ser Ile Ile Val Val Phe Asp Leu Phe Phe Ala
115 120 125
Gln Trp Val Ser Asp Gln Asn Val Lys Glu Glu Leu Ile Gln Gly Leu
130 135 140
Glu Ala Asn Lys Ser Ser Gln Leu Val Thr Phe His Ile Asp Leu Asn
145 150 155 160
Ser Val Asp Ile Leu Asp Lys Leu Thr Thr Thr Ser His Leu Ala Thr
165 170 175
Pro Gly Asn Val Ser Ile Glu Cys Leu Pro Gly Ser Ser Pro Cys Thr
180 185 190
Asp Ala Leu Thr Cys Ile Lys Ala Asp Leu Phe Cys Asp Gly Glu Val
195 200 205
Asn Cys Pro Asp Gly Ser Asp Glu Asp Asn Lys Met Cys Ala Thr Val
210 215 220
Cys Asp Gly Arg Phe Leu Leu Thr Gly Ser Ser Gly Ser Phe Gln Ala
225 230 235 240
Thr His Tyr Pro Lys Pro Ser Glu Thr Ser Val ValCys Gln Trp Ile
245 250 255
Ile Arg Val Asn Gln Gly Leu Ser Ile Lys Leu Ser Phe Asp Asp Phe
260 265 270
Asn Thr Tyr Tyr Thr Asp Ile Leu Asp Ile Tyr Glu Gly Val Gly Ser
275 280 285
Ser Lys Ile Leu Arg Ala Ser Ile Trp Glu Thr Asn Pro Gly Thr Ile
290 295 300
Arg Ile Phe Ser Asn Gln Val Thr Ala Thr Phe Leu Ile Glu Ser Asp
305 310 315 320
Glu Ser Asp Tyr Val Gly Phe Asn Ala Thr Tyr Thr Ala Phe Asn Ser
325 330 335
Ser Glu Leu Asn Asn Tyr Glu Lys Ile Asn Cys Asn Phe Glu Asp Gly
340 345 350
Phe Cys Phe Trp Val Gln Asp Leu Asn Asp Asp Asn Glu Trp Glu Arg
355 360 365
Ile Gln Gly Ser Thr Phe Ser Pro Phe Thr Gly Pro Asn Phe Asp His
370 375 380
Thr Phe Gly Asn Ala Ser Gly Phe Tyr Ile Ser Thr Pro Thr Gly Pro
385 390 395 400
Gly Gly Arg Gln Glu Arg Val Gly Leu Leu Ser Leu Pro Leu Asp Pro
405 410 415
Thr Leu Glu Pro Ala Cys Leu Ser Phe Trp Tyr His Met Tyr Gly Glu
420 425 430
Asn Val His Lys Leu Ser Ile Asn Ile Ser Asn Asp Gln Asn Met Glu
435 440 445
Lys Thr Val Phe Gln Lys Glu Gly Asn Tyr Gly Asp Asn Trp Asn Tyr
450 455 460
Gly Gln Val Thr Leu Asn Glu Thr Val Lys Phe Lys Val Ala Phe Asn
465 470 475 480
Ala Phe Lys Asn Lys Ile Leu Ser Asp Ile Ala Leu Asp Asp Ile Ser
485 490 495
Leu Thr Tyr Gly Ile Cys Asn Gly Ser Leu Tyr Pro Glu Pro Thr Leu
500 505 510
Val Pro Thr Pro Pro Pro Glu Leu Pro Thr Asp Cys Gly Gly Pro Phe
515 520 525
Glu Leu Trp Glu Pro Asn Thr Thr Phe Ser Ser Thr Asn Phe Pro Asn
530 535 540
Ser Tyr Pro Asn Leu Ala Phe Cys Val Trp Ile Leu Asn Ala Gln Lys
545 550 555 560
Gly Lys Asn Ile Gln Leu His Phe Gln Glu Phe Asp Leu Glu Asn Ile
565 570 575
Asn Asp Val Val Glu Ile Arg Asp Gly Glu Glu Ala Asp Ser Leu Leu
580 585 590
Leu Ala Val Tyr Thr Gly Pro Gly Pro Val Lys Asp Val Phe Ser Thr
595 600 605
Thr Asn Arg Met Thr Val Leu Leu Ile Thr Asn Asp Val Leu Ala Arg
610 615 620
Gly Gly Phe Lys Ala Asn Phe Thr Thr Gly Tyr His Leu Gly Ile Pro
625 630 635 640
Glu Pro Cys Lys Ala Asp His Phe Gln Cys Lys Asn Gly Glu Cys Val
645 650 655
Pro Leu Val Asn Leu Cys Asp Gly His Leu His Cys Glu Asp Gly Ser
660 665 670
Asp Glu Ala Asp Cys Val Arg Phe Phe Asn Gly Thr Thr Asn Asn Asn
675 680 685
Gly Leu Val Arg Phe Arg Ile Gln Ser Ile Trp His Thr Ala Cys Ala
690 695 700
Glu Asn Trp Thr Thr Gln Ile Ser Asn Asp Val Cys Gln Leu Leu Gly
705 710 715 720
Leu Gly Ser Gly Asn Ser Ser Lys Pro Ile Phe Ser Thr Asp Gly Gly
725 730 735
Pro Phe Val Lys Leu Asn Thr Ala Pro Asp Gly His Leu Ile Leu Thr
740 745 750
Pro Ser Gln Gln Cys Leu Gln Asp Ser Leu Ile Arg Leu Gln Cys Asn
755 760 765
His Lys Ser Cys Gly Lys Lys Leu Ala Ala Gln Asp Ile Thr Pro Lys
770 775 780
Ile Val Gly Gly Ser Asn Ala Lys Glu Gly Ala Trp Pro Trp Val Val
785 790 795 800
Gly Leu Tyr Tyr Gly Gly Arg Leu Leu Cys Gly Ala Ser Leu Val Ser
805 810 815
Ser Asp Trp Leu Val Ser Ala Ala His Cys Val Tyr Gly Arg Asn Leu
820 825 830
Glu Pro Ser Lys Trp Thr Ala Ile Leu Gly Leu His Met Lys Ser Asn
835 840 845
Leu Thr Ser Pro Gln Thr Val Pro Arg Leu Ile Asp Glu Ile Val Ile
850 855 860
Asn Pro His Tyr Asn Arg Arg Arg Lys Asp Asn Asp Ile Ala Met Met
865 870 875 880
His Leu Glu Phe Lys Val Asn Tyr Thr Asp Tyr Ile Gln Pro Ile Cys
885 890 895
Leu Pro Glu Glu Asn Gln Val Phe Pro Pro Gly Arg Asn Cys Ser Ile
900 905 910
Ala Gly Trp Gly Thr Val Val Tyr Gln Gly Thr Thr Ala Asn Ile Leu
915 920 925
Gln Glu Ala Asp Val Pro Leu Leu Ser Asn Glu Arg Cys Gln Gln Gln
930 935 940
Met Pro Glu Tyr Asn Ile Thr Glu Asn Met Ile Cys Ala Gly Tyr Glu
945 950 955 960
Glu Gly Gly Ile Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Met
965 970 975
Cys Gln Glu Asn Asn Arg Trp Phe Leu Ala Gly Val Thr Ser Phe Gly
980 985 990
Tyr Lys Cys Ala Leu Pro Asn Arg Pro Gly Val Tyr Ala Arg Val Ser
995 1000 1005
Arg Phe Thr Glu Trp Ile Gln Ser Phe Leu His
1010 1015
<210>134
<211>917
<212>DNA
<213>Homo sapiens
<220>
<223>chymotrypsin C
<400>134
cagccagtcc tgagcaccta accatgttgg gcatcactgt cctcgctgcg ctcttggcct 60
gtgcctccag ctgtggggtg cccagcttcc cgcccaacct atccgcccga gtggtgggag 120
gagaggatgc ccggccccac agctggccct ggcagatctc cctccagtac ctcaagaacg 180
acacgtggag gcatacgtgt ggcgggactt tgattgctag caacttcgtc ctcactgccg 240
cccactgcat cagcaacacc cggacctacc gtgtggccgt gggaaagaac aacctggagg 300
tggaagacga agaaggatcc ctgtttgtgg gtgtggacac catccacgtc cacaagagat 360
ggaatgccct cctgttgcgc aatgatattg ccctcatcaa gcttgcagag catgtggagc 420
tgagtgacac catccaggtg gcctgcctgc cagagaagga ctccctgctc cccaaggact 480
acccctgcta tgtcaccggc tggggccgcc tctggaccaa cggccccatt gctgataagc 540
tgcagcaggg cctgcagccc gtggtggatc acgccacgtg ctccaggatt gactggtggg 600
gcttcagggt gaagaaaacc atggtgtgcg ctgggggcga tggcgtcatc tcagcctgca 660
atggggactc cggtggccca ctgaactgcc agttggagaa cggttcctgg gaggtgtttg 720
gcatcgtcag ctttggctcc cggcggggct gcaacacccg caagaagccg gtagtctaca 780
cccgggtgtc cgcctacatc gactggatca acgagaaaat gcagctgtga tttgttgctg 840
ggagcggcgg cagcgagtcc ctgcaacagc aataaacttc cttctcctcg ggccacctga 900
aaaaaaaaaa aaaaaaa 917
<210>135
<211>268
<212>PRT
<213>Homo sapiens
<220>
<223>chymotrypsin C
<400>135
Met Leu Gly Ile Thr Val Leu Ala Ala Leu Leu Ala Cys Ala Ser Ser
1 5 10 15
Cys Gly Val Pro Ser Phe Pro Pro Asn Leu Ser Ala Arg Val Val Gly
20 25 30
Gly Glu Asp Ala Arg Pro His Ser Trp Pro Trp Gln Ile Ser Leu Gln
35 40 45
Tyr Leu Lys Asn Asp Thr Trp Arg His Thr Cys Gly Gly Thr Leu Ile
50 55 60
Ala Ser Asn Phe Val Leu Thr Ala Ala His Cys Ile Ser Asn Thr Arg
65 70 75 80
Thr Tyr Arg Val Ala Val Gly Lys Asn Asn Leu Glu Val Glu Asp Glu
85 90 95
Glu Gly Ser Leu Phe Val Gly Val Asp Thr Ile His Val His Lys Arg
100 105 110
Trp Asn Ala Leu Leu Leu Arg Asn Asp Ile Ala Leu Ile Lys Leu Ala
115 120 125
Glu His Val Glu Leu Ser Asp Thr Ile Gln Val Ala Cys Leu Pro Glu
130 135 140
Lys Asp Ser Leu Leu Pro Lys Asp Tyr Pro Cys Tyr Val Thr Gly Trp
145 150 155 160
Gly Arg Leu Trp Thr Asn Gly Pro Ile Ala Asp Lys Leu Gln Gln Gly
165 170 175
Leu Gln Pro Val Val Asp His Ala Thr Cys Ser Arg Ile Asp Trp Trp
180 185 190
Gly Phe Arg Val Lys Lys Thr Met Val Cys Ala Gly Gly Asp Gly Val
195 200 205
Ile Ser Ala Cys Asn Gly Asp Ser Gly Gly Pro Leu Asn Cys Gln Leu
210 215 220
Glu Asn Gly Ser Trp Glu Val Phe Gly Ile Val Ser Phe Gly Ser Arg
225 230 235 240
Arg Gly Cys Asn Thr Arg Lys Lys Pro Val Val Tyr Thr Arg Val Ser
245 250 255
Ala Tyr Ile Asp Trp Ile Asn Glu Lys Met Gln Leu
260 265
<210>136
<211>1938
<212>DNA
<213>Homo sapoens
<220>
<223>Prostasin
<400>136
gactttggtg gcaagaggag ctggcggagc ccagccagtg ggcggggcca ggggaggggc 60
gggcaggtag gtgcagccac tcctgggagg accctgcgtg gccagacggt gctggtgact 120
cgtccacact gctcgcttcg gatactccag gcgtctcccg ttgcggccgc tccctgcctt 180
agaggccagc cttggacact tgctgcccct ttccagcccg gattctggga tccttccctc 240
tgagccaaca tctgggtcct gccttcgaca ccaccccaag gcttcctacc ttgcgtgcct 300
ggagtctgcc ccaggggccc ttgtcctggg ccatggccca gaagggggtc ctggggcctg 360
ggcagctggg ggctgtggcc attctgctct atcttggatt actccggtcg gggacaggag 420
cggaaggggc agaagctccc tgcggtgtgg ccccccaagc acgcatcaca ggtggcagca 480
gtgcagtcgc cggtcagtgg ccctggcagg tcagcatcac ctatgaaggc gtccatgtgt 540
gtggtggctc tctcgtgtct gagcagtggg tgctgtcagc tgctcactgc ttccccagcg 600
agcaccacaa ggaagcctat gaggtcaagc tgggggccca ccagctagac tcctactccg 660
aggacgccaa ggtcagcacc ctgaaggaca tcatccccca ccccagctac ctccaggagg 720
gctcccaggg cgacattgca ctcctccaac tcagcagacc catcaccttc tcccgctaca 780
tccggcccat ctgcctccct gcagccaacg cctccttccc caacggcctc cactgcactg 840
tcactggctg gggtcatgtg gccccctcag tgagcctcct gacgcccaag ccactgcagc 900
aactcgaggt gcctctgatc agtcgtgaga cgtgtaactg cctgtacaac atcgacgcca 960
agcctgagga gccgcacttt gtccaagagg acatggtgtg tgctggctat gtggaggggg 1020
gcaaggacgc ctgccagggt gactctgggg gcccactctc ctgccctgtg gagggtctct 1080
ggtacctgac gggcattgtg agctggggag atgcctgtgg ggcccgcaac aggcctggtg 1140
tgtacactct ggcctccagc tatgcctcct ggatccaaag caaggtgaca gaactccagc 1200
ctcgtgtggt gccccaaacc caggagtccc agcccgacag caacctctgt ggcagccacc 1260
tggccttcag ctctgcccca gcccagggct tgctgaggcc catccttttc ctgcctctgg 1320
gcctggctct gggcctcctc tccccatggc tcagcgagca ctgagctggc cctacttcca 1380
ggatggatgc atcacactca aggacaggag cctggtcctt ccctgatggc ctttggaccc 1440
agggcctgac ttgagccact ccttccttca ggactctgcg ggaggctggg gccccatctt 1500
gatctttgag cccattcttc tgggtgtgct ttttgggacc atcactgaga gtcaggagtt 1560
ttactgcctg tagcaatggc cagagcctct ggcccctcac ccaccatgga ccagcccatt 1620
ggccgagctc ctggggagct cctgggaccc ttggctatga aaatgagccc tggctcccac 1680
ctgtttctgg aagactgctc ccggcccgcc tgcccagact gatgagcaca tctctctgcc 1740
ctctccctgt gttctgggct ggggccacct ttgtgcagct tcgaggacag gaaaggcccc 1800
aatcttgccc actggccgct gagcgccccc gagccctgac tcctggactc cggaggactg 1860
agcccccacc ggaactgggc tggcgcttgg atctggggtg ggagtaacag ggcagaaatg 1920
attaaaatgt ttgagcac 1938
<210>137
<211>343
<212>PRT
<213>Homo sapiens
<220>
<223>Prostasin
<400>137
Met Ala Gln Lys Gly Val Leu Gly Pro Gly Gln Leu Gly Ala Val Ala
1 5 10 15
Ile Leu Leu Tyr Leu Gly Leu Leu Arg Ser Gly Thr Gly Ala Glu Gly
20 25 30
Ala Glu Ala Pro Cys Gly Val Ala Pro Gln Ala Arg Ile Thr Gly Gly
35 40 45
Ser Ser Ala Val Ala Gly Gln Trp Pro Trp Gln Val Ser Ile Thr Tyr
50 55 60
Glu Gly Val His Val Cys Gly Gly Ser Leu Val Ser Glu Gln Trp Val
65 70 75 80
Leu Ser Ala Ala His Cys Phe Pro Ser Glu His His Lys Glu Ala Tyr
85 90 95
Glu Val Lys Leu Gly Ala His Gln Leu Asp Ser Tyr Ser Glu Asp Ala
100 105 110
Lys Val Ser Thr Leu Lys Asp Ile Ile Pro His Pro Ser Tyr Leu Gln
115 120 125
Glu Gly Ser Gln Gly Asp Ile Ala Leu Leu Gln Leu Ser Arg Pro Ile
130 135 140
Thr Phe Ser Arg Tyr Ile Arg Pro Ile Cys Leu Pro Ala Ala Asn Ala
145 150 155 160
Ser Phe Pro Asn Gly Leu His Cys Thr Val Thr Gly Trp Gly His Val
165 170 175
Ala Pro Ser Val Ser Leu Leu Thr Pro Lys Pro Leu Gln Gln Leu Glu
180 185 190
Val Pro Leu Ile Ser Arg Glu Thr Cys Asn Cys Leu Tyr Asn Ile Asp
195 200 205
Ala Lys Pro Glu Glu Pro His Phe Val Gln Glu Asp Met Val Cys Ala
210 215 220
Gly Tyr Val Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp Ser Gly Gly
225 230 235 240
Pro Leu Ser Cys Pro Val Glu Gly Leu Trp Tyr Leu Thr Gly Ile Val
245 250 255
Ser Trp Gly Asp Ala Cys Gly Ala Arg Asn Arg Pro Gly Val Tyr Thr
260 265 270
Leu Ala Ser Ser Tyr Ala Ser Trp Ile Gln Ser Lys Val Thr Glu Leu
275 280 285
Gln Pro Arg Val Val Pro Gln Thr Gln Glu Ser Gln Pro Asp Ser Asn
290 295 300
Leu Cys Gly Ser His Leu Ala Phe Ser Ser Ala Pro Ala Gln Gly Leu
305 310 315 320
Leu Arg Pro Ile Leu Phe Leu Pro Leu Gly Leu Ala Leu Gly Leu Leu
325 330 335
Ser Pro Trp Leu Ser Glu His
340
<210>138
<211>874
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein 1
<400>138
agttcctcca cctgctggcc cctggacacc tctgtcacca tgtggttcct ggttctgtgc 60
ctcgccctgt ccctgggggg gactggtgct gcgcccccga ttcagtcccg gattgtggga 120
ggctgggagt gtgagcagca ttcccagccc tggcaggcgg ctctgtacca tttcagcact 180
ttccagtgtg ggggcatcct ggtgcaccgc cagtgggtgc tcacagctgc tcattgcatc 240
agcgacaatt accagctctg gctgggtcgc cacaacttgt ttgacgacga aaacacagcc 300
cagtttgttc atgtcagtga gagcttccca caccctggct tcaacatgag cctcctggag 360
aaccacaccc gccaagcaga cgaggactac agccacgacc tcatgctgct ccgcctgaca 420
gagcctgctg ataccatcac agatgctgtg aaggtcgtgg agttgcccac cgaggaaccc 480
gaagtgggga gcacctgttt ggcttccggc tggggcagca tcgaaccaga gaatttctca 540
tttccagatg atctccagtg tgtggacctc aaaatcctgc ctaatgatga gtgcaaaaaa 600
gcccacgtcc agaaggtgac agacttcatg ctgtgtgtcg gacacctgga aggtggcaaa 660
gacacctgtg tgggtgattc agggggcccg ctgatgtgtg atggtgtgct ccaaggtgtc 720
acatcatggg gctacgtccc ttgtggcacc cccaataagc cttctgtcgc cgtcagagtg 780
ctgtcttatg tgaagtggat cgaggacacc atagcggaga actcctgaac gcccagccct 840
gtcccctacc cccagtaaaa tcaaatgtgc atcc 874
<210>139
<211>262
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein 1
<400>139
Met Trp Phe Leu Val Leu Cys Leu Ala Leu Ser Leu Gly Gly Thr Gly
1 5 10 15
Ala Ala Pro Pro Ile Gln Ser Arg Ile Val Gly Gly Trp Glu Cys Glu
20 25 30
Gln His Ser Gln Pro Trp Gln Ala Ala Leu Tyr His Phe Ser Thr Phe
35 40 45
Gln Cys Gly Gly Ile Leu Val His Arg Gln Trp Val Leu Thr Ala Ala
50 55 60
His Cys Ile Ser Asp Asn Tyr Gln Leu Trp Leu Gly Arg His Asn Leu
65 70 75 80
Phe Asp Asp Glu Asn Thr Ala Gln Phe Val His Val Ser Glu Ser Phe
85 90 95
Pro His Pro Gly Phe Asn Met Ser Leu Leu Glu Asn His Thr Arg Gln
100 105 110
Ala Asp Glu Asp Tyr Ser His Asp Leu Met Leu Leu Arg Leu Thr Glu
115 120 125
Pro Ala Asp Thr Ile Thr Asp Ala Val Lys Val Val Glu Leu Pro Thr
130 135 140
Glu Glu Pro Glu Val Gly Ser Thr Cys Leu Ala Ser Gly Trp Gly Ser
145 150 155 160
Ile Glu Pro Glu Asn Phe Ser Phe Pro Asp Asp Leu Gln Cys Val Asp
165 170 175
Leu Lys Ile Leu Pro Asn Asp Glu Cys Lys Lys Ala His Val Gln Lys
180 185 190
Val Thr Asp Phe Met Leu Cys Val Gly His Leu Glu Gly Gly Lys Asp
195 200 205
Thr Cys Val Gly Asp Ser Gly Gly Pro Leu Met Cys Asp Gly Val Leu
210 215 220
Gln Gly Val Thr Ser Trp Gly Tyr Val Pro Cys Gly Thr Pro Asn Lys
225 230 235 240
Pro Ser Val Ala Val Arg Val Leu Ser Tyr Val Lys Trp Ile Glu Asp
245 250 255
Thr Ile Ala Glu Asn Ser
260
<210>140
<211>2855
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK2 transcript variant 1
<400>140
agccccaaac tcaccacctg gccgtggaca cctgtgtcag catgtgggac ctggttctct 60
ccatcgcctt gtctgtgggg tgcactggtg ccgtgcccct catccagtct cggattgtgg 120
gaggctggga gtgtgagaag cattcccaac cctggcaggt ggctgtgtac agtcatggat 180
gggcacactg tgggggtgtc ctggtgcacc cccagtgggt gctcacagct gcccattgcc 240
taaagaagaa tagccaggtc tggctgggtc ggcacaacct gtttgagcct gaagacacag 300
gccagagggt ccctgtcagc cacagcttcc cacacccgct ctacaatatg agccttctga 360
agcatcaaag ccttagacca gatgaagact ccagccatga cctcatgctg ctccgcctgt 420
cagagcctgc caagatcaca gatgttgtga aggtcctggg cctgcccacc caggagccag 480
cactggggac cacctgctac gcctcaggct ggggcagcat cgaaccagag gagttcttgc 540
gccccaggag tcttcagtgt gtgagcctcc atctcctgtc caatgacatg tgtgctagag 600
cttactctga gaaggtgaca gagttcatgt tgtgtgctgg gctctggaca ggtggtaaag 660
acacttgtgg gggtgattct gggggtccac ttgtctgtaa tggtgtgctt caaggtatca 720
catcatgggg ccctgagcca tgtgccctgc ctgaaaagcc tgctgtgtac accaaggtgg 780
tgcattaccg gaagtggatc aaggacacca tcgcagccaa cccctgagtg cccctgtccc 840
acccctacct ctagtaaatt taagtccacc tcacgttctg gcatcacttg gcctttctgg 900
atgctggaca cctgaagctt ggaactcacc tggccgaagc tcgagcctcc tgagtcctac 960
tgacctgtgc tttctggtgt ggagtccagg gctgctagga aaaggaatgg gcagacacag 1020
gtgtatgcca atgtttctga aatgggtata atttcgtcct ctccttcgga acactggctg 1080
tctctgaaga cttctcgctc agtttcagtg aggacacaca caaagacgtg ggtgaccatg 1140
ttgtttgtgg ggtgcagaga tgggaggggt ggggcccacc ctggaagagt ggacagtgac 1200
acaaggtgga cactctctac agatcactga ggataagctg gagccacaat gcatgaggca 1260
cacacacagc aaggatgacg ctgtaaacat agcccacgct gtcctggggg cactgggaag 1320
cctagataag gccgtgagca gaaagaaggg gaggatcctc ctatgttgtt gaaggaggga 1380
ctagggggag aaactgaaag ctgattaatt acaggaggtt tgttcaggtc ccccaaacca 1440
ccgtcagatt tgatgatttc ctagcaggac ttacagaaat aaagagctat catgctgtgg 1500
tttattatgg tttgttacat tgatgggata catactgaaa tcagcaaaca aaacagatgt 1560
atagattaga gtgtggagaa aacagaggaa aacttgcagt tacgaagact ggcaacttgg 1620
ctttactaag ttttcagact ggcaggaagt caaacctatt aggctgagga ccttgtggag 1680
tgtagctgat ccagctgata gaggaactag ccaggtgggg gcctttccct ttggatgggg 1740
ggcatatctg acagttattc tctccaagtg gagacttacg gacagcatat aattctccct 1800
gcaaggatgt atgataatat gtacaaagta attccaactg aggaagctca cctgatcctt 1860
agtgtccaag gtttttactg ggggtctgta ggacgagtat ggagtacttg aataattgac 1920
ctgaagtcct cagacctgag gttccctaga gttcaaacag atacagcatg gtccagagtc 1980
ccagatgtac aaaaacaggg attcatcaca aatcccatct ttagcatgaa gggtctggca 2040
tggcccaagg ccccaagtat atcaaggcac ttgggcagaa catgccaagg aatcaaatgt 2100
catctcccag gagttattca agggtgagcc ctttacttgg gatgtacagg ctttgagcag 2160
tgcagggctg ctgagtcaac cttttattgt acaggggatg agggaaaggg agaggatgag 2220
gaagcccccc tggggatttg gtttggtctt gtgatcaggt ggtctatggg gctatcccta 2280
caaagaagaa tccagaaata ggggcacatt gaggaatgat actgagccca aagagcattc 2340
aatcattgtt ttatttgcct tcttttcaca ccattggtga gggagggatt accaccctgg 2400
ggttatgaag atggttgaac accccacaca tagcaccgga gatatgagat caacagtttc 2460
ttagccatag agattcacag cccagagcag gaggacgctg cacaccatgc aggatgacat 2520
gggggatgcg ctcgggattg gtgtgaagaa gcaaggactg ttagaggcag gctttatagt 2580
aacaagacgg tggggcaaac tctgatttcc gtgggggaat gtcatggtct tgctttacta 2640
agttttgaga ctggcaggta gtgaaactca ttaggctgag aaccttgtgg aatgcagctg 2700
acccagctga tagaggaagt agccaggtgg gagcctttcc cagtgggtgt gggacatatc 2760
tggcaagatt ttgtggcact cctggttaca gatactgggg cagcaaataa aactgaatct 2820
tgttttcaga ccttaaaaaa aaaaaaaaaa aaaaa 2855
<210>141
<211>261
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hK2 isoform 1
<400>141
Met Trp Asp Leu Val Leu Ser Ile Ala Leu Ser Val Gly Cys Thr Gly
1 5 10 15
Ala Val Pro Leu Ile Gln Ser Arg Ile Val Gly Gly Trp Glu Cys Glu
20 25 30
Lys His Ser Gln Pro Trp Gln Val Ala Val Tyr Ser His Gly Trp Ala
35 40 45
His Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala
50 55 60
His Cys Leu Lys Lys Asn Ser Gln Val Trp Leu Gly Arg His Asn Leu
65 70 75 80
Phe Glu Pro Glu Asp Thr Gly Gln Arg Val Pro Val Ser His Ser Phe
85 90 95
Pro His Pro Leu Tyr Asn Met Ser Leu Leu Lys His Gln Ser Leu Arg
100 105 110
Pro Asp Glu Asp Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser Glu
115 120 125
Pro Ala Lys Ile Thr Asp Val Val Lys Val Leu Gly Leu Pro Thr Gln
130 135 140
Glu Pro Ala Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser Ile
145 150 155 160
Glu Pro Glu Glu Phe Leu Arg Pro Arg Ser Leu Gln Cys Val Ser Leu
165 170 175
His Leu Leu Ser Asn Asp Met Cys Ala Arg Ala Tyr Ser Glu Lys Val
180 185 190
Thr Glu Phe Met Leu Cys Ala Gly Leu Trp Thr Gly Gly Lys Asp Thr
195 200 205
Cys Gly Gly Asp Ser Gly Gly Pro Leu Val Cys Asn Gly Val Leu Gln
210 215 220
Gly Ile Thr Ser Trp Gly Pro Glu Pro Cys Ala Leu Pro Glu Lys Pro
225 230 235 240
Ala Val Tyr Thr Lys Val Val His Tyr Arg Lys Trp Ile Lys Asp Thr
245 250 255
Ile Ala Ala Asn Pro
260
<210>142
<211>2892
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK2 transcript variant 2
<400>142
agccccaaac tcaccacctg gccgtggaca cctgtgtcag catgtgggac ctggttctct 60
ccatcgcctt gtctgtgggg tgcactggtg ccgtgcccct catccagtct cggattgtgg 120
gaggctggga gtgtgagaag cattcccaac cctggcaggt ggctgtgtac agtcatggat 180
gggcacactg tgggggtgtc ctggtgcacc cccagtgggt gctcacagct gcccattgcc 240
taaagaagaa tagccaggtc tggctgggtc ggcacaacct gtttgagcct gaagacacag 300
gccagagggt ccctgtcagc cacagcttcc cacacccgct ctacaatatg agccttctga 360
agcatcaaag ccttagacca gatgaagact ccagccatga cctcatgctg ctccgcctgt 420
cagagcctgc caagatcaca gatgttgtga aggtcctggg cctgcccacc caggagccag 480
cactggggac cacctgctac gcctcaggct ggggcagcat cgaaccagag gagttcttgc 540
gccccaggag tcttcagtgt gtgagcctcc atctcctgtc caatgacatg tgtgctagag 600
cttactctga gaaggtgaca gagttcatgt tgtgtgctgg gctctggaca ggtggtaaag 660
acacttgtgg ggtgagtcat ccctactccc aacatctgga ggggaaaggg tgattctggg 720
ggtccacttg tctgtaatgg tgtgcttcaa ggtatcacat catggggccc tgagccatgt 780
gccctgcctg aaaagcctgc tgtgtacacc aaggtggtgc attaccggaa gtggatcaag 840
gacaccatcg cagccaaccc ctgagtgccc ctgtcccacc cctacctcta gtaaatttaa 900
gtccacctca cgttctggca tcacttggcc tttctggatg ctggacacct gaagcttgga 960
actcacctgg ccgaagctcg agcctcctga gtcctactga cctgtgcttt ctggtgtgga 1020
gtccagggct gctaggaaaa ggaatgggca gacacaggtg tatgccaatg tttctgaaat 1080
gggtataatt tcgtcctctc cttcggaaca ctggctgtct ctgaagactt ctcgctcagt 1140
ttcagtgagg acacacacaa agacgtgggt gaccatgttg tttgtggggt gcagagatgg 1200
gaggggtggg gcccaccctg gaagagtgga cagtgacaca aggtggacac tctctacaga 1260
tcactgagga taagctggag ccacaatgca tgaggcacac acacagcaag gatgacgctg 1320
taaacatagc ccacgctgtc ctgggggcac tgggaagcct agataaggcc gtgagcagaa 1380
agaaggggag gatcctccta tgttgttgaa ggagggacta gggggagaaa ctgaaagctg 1440
attaattaca ggaggtttgt tcaggtcccc caaaccaccg tcagatttga tgatttccta 1500
gcaggactta cagaaataaa gagctatcat gctgtggttt attatggttt gttacattga 1560
tgggatacat actgaaatca gcaaacaaaa cagatgtata gattagagtg tggagaaaac 1620
agaggaaaac ttgcagttac gaagactggc aacttggctt tactaagttt tcagactggc 1680
aggaagtcaa acctattagg ctgaggacct tgtggagtgt agctgatcca gctgatagag 1740
gaactagcca ggtgggggcc tttccctttg gatggggggc atatctgaca gttattctct 1800
ccaagtggag acttacggac agcatataat tctccctgca aggatgtatg ataatatgta 1860
caaagtaatt ccaactgagg aagctcacct gatccttagt gtccaaggtt tttactgggg 1920
gtctgtagga cgagtatgga gtacttgaat aattgacctg aagtcctcag acctgaggtt 1980
ccctagagtt caaacagata cagcatggtc cagagtccca gatgtacaaa aacagggatt 2040
catcacaaat cccatcttta gcatgaaggg tctggcatgg cccaaggccc caagtatatc 2100
aaggcacttg ggcagaacat gccaaggaat caaatgtcat ctcccaggag ttattcaagg 2160
gtgagccctt tacttgggat gtacaggctt tgagcagtgc agggctgctg agtcaacctt 2220
ttattgtaca ggggatgagg gaaagggaga ggatgaggaa gcccccctgg ggatttggtt 2280
tggtcttgtg atcaggtggt ctatggggct atccctacaa agaagaatcc agaaataggg 2340
gcacattgag gaatgatact gagcccaaag agcattcaat cattgtttta tttgccttct 2400
tttcacacca ttggtgaggg agggattacc accctggggt tatgaagatg gttgaacacc 2460
ccacacatag caccggagat atgagatcaa cagtttctta gccatagaga ttcacagccc 2520
agagcaggag gacgctgcac accatgcagg atgacatggg ggatgcgctc gggattggtg 2580
tgaagaagca aggactgtta gaggcaggct ttatagtaac aagacggtgg ggcaaactct 2640
gatttccgtg ggggaatgtc atggtcttgc tttactaagt tttgagactg gcaggtagtg 2700
aaactcatta ggctgagaac cttgtggaat gcagctgacc cagctgatag aggaagtagc 2760
caggtgggag cctttcccag tgggtgtggg acatatctgg caagattttg tggcactcct 2820
ggttacagat actggggcag caaataaaac tgaatcttgt tttcagacct taaaaaaaaa 2880
aaaaaaaaaa aa 2892
<210>143
<211>223
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hK2 isoform 2
<400>143
Met Trp Asp Leu Val Leu Ser Ile Ala Leu Ser Val Gly Cys Thr Gly
1 5 10 15
Ala Val Pro Leu Ile Gln Ser Arg Ile Val Gly Gly Trp Glu Cys Glu
20 25 30
Lys His Ser Gln Pro Trp Gln Val Ala Val Tyr Ser His Gly Trp Ala
35 40 45
His Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala
50 55 60
His Cys Leu Lys Lys Asn Ser Gln Val Trp Leu Gly Arg His Asn Leu
65 70 75 80
Phe Glu Pro Glu Asp Thr Gly Gln Arg Val Pro Val Ser His Ser Phe
85 90 95
Pro His Pro Leu Tyr Asn Met Ser Leu Leu Lys His Gln Ser Leu Arg
100 105 110
Pro Asp Glu Asp Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser Glu
115 120 125
Pro Ala Lys lle Thr Asp Val Val Lys Val Leu Gly Leu Pro Thr Gln
130 135 140
Glu Pro Ala Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser Ile
145 150 155 160
Glu Pro Glu Glu Phe Leu Arg Pro Arg Ser Leu Gln Cys Val Ser Leu
165 170 175
His Leu Leu Ser Asn Asp Met Cys Ala Arg Ala Tyr Ser Glu Lys Val
180 185 190
Thr Glu Phe Met Leu Cys Ala Gly Leu Trp Thr Gly Gly Lys Asp Thr
195 200 205
Cys Gly Val Ser His Pro Tyr Ser Gln His Leu Glu Gly Lys Gly
210 215 220
<210>144
<211>2879
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK2 transcript variant 3
<400>144
agccccaaac tcaccacctg gccgtggaca cctgtgtcag catgtgggac ctggttctct 60
ccatcgcctt gtctgtgggg tgcactggtg ccgtgcccct catccagtct cggattgtgg 120
gaggctggga gtgtgagaag cattcccaac cctggcaggt ggctgtgtac agtcatggat 180
gggcacactg tgggggtgtc ctggtgcacc cccagtgggt gctcacagct gcccattgcc 240
taaagaagaa tagccaggtc tggctgggtc ggcacaacct gtttgagcct gaagacacag 300
gccagagggt ccctgtcagc cacagcttcc cacacccgct ctacaatatg agccttctga 360
agcatcaaag ccttagacca gatgaagact ccagccatga cctcatgctg ctccgcctgt 420
cagagcctgc caagatcaca gatgttgtga aggtcctggg cctgcccacc caggagccag 480
cactggggac cacctgctac gcctcaggct ggggcagcat cgaaccagag gagtgagtct 540
tcagtgtgtg agcctccatc tcctgtccaa tgacatgtgt gctagagctt actctgagaa 600
ggtgacagag ttcatgttgt gtgctgggct ctggacaggt ggtaaagaca cttgtggggt 660
gagtcatccc tactcccaac atctggaggg gaaagggtga ttctgggggt ccacttgtct 720
gtaatggtgt gcttcaaggt atcacatcat ggggccctga gccatgtgcc ctgcctgaaa 780
agcctgctgt gtacaccaag gtggtgcatt accggaagtg gatcaaggac accatcgcag 840
ccaacccctg agtgcccctg tcccacccct acctctagta aatttaagtc cacctcacgt 900
tctggcatca cttggccttt ctggatgctg gacacctgaa gcttggaact cacctggccg 960
aagctcgagc ctcctgagtc ctactgacct gtgctttctg gtgtggagtc cagggctgct 1020
aggaaaagga atgggcagac acaggtgtat gccaatgttt ctgaaatggg tataatttcg 1080
tcctctcctt cggaacactg gctgtctctg aagacttctc gctcagtttc agtgaggaca 1140
cacacaaaga cgtgggtgac catgttgttt gtggggtgca gagatgggag gggtggggcc 1200
caccctggaa gagtggacag tgacacaagg tggacactct ctacagatca ctgaggataa 1260
gctggagcca caatgcatga ggcacacaca cagcaaggat gacgctgtaa acatagccca 1320
cgctgtcctg ggggcactgg gaagcctaga taaggccgtg agcagaaaga aggggaggat 1380
cctcctatgt tgttgaagga gggactaggg ggagaaactg aaagctgatt aattacagga 1440
ggtttgttca ggtcccccaa accaccgtca gatttgatga tttcctagca ggacttacag 1500
aaataaagag ctatcatgct gtggtttatt atggtttgtt acattgatgg gatacatact 1560
gaaatcagca aacaaaacag atgtatagat tagagtgtgg agaaaacaga ggaaaacttg 1620
cagttacgaa gactggcaac ttggctttac taagttttca gactggcagg aagtcaaacc 1680
tattaggctg aggaccttgt ggagtgtagc tgatccagct gatagaggaa ctagccaggt 1740
gggggccttt ccctttggat ggggggcata tctgacagtt attctctcca agtggagact 1800
tacggacagc atataattct ccctgcaagg atgtatgata atatgtacaa agtaattcca 1860
actgaggaag ctcacctgat ccttagtgtc caaggttttt actgggggtc tgtaggacga 1920
gtatggagta cttgaataat tgacctgaag tcctcagacc tgaggttccc tagagttcaa 1980
acagatacag catggtccag agtcccagat gtacaaaaac agggattcat cacaaatccc 2040
atctttagca tgaagggtct ggcatggccc aaggccccaa gtatatcaag gcacttgggc 2100
agaacatgcc aaggaatcaa atgtcatctc ccaggagtta ttcaagggtg agccctttac 2160
ttgggatgta caggctttga gcagtgcagg gctgctgagt caacctttta ttgtacaggg 2220
gatgagggaa agggagagga tgaggaagcc cccctgggga tttggtttgg tcttgtgatc 2280
aggtggtcta tggggctatc cctacaaaga agaatccaga aataggggca cattgaggaa 2340
tgatactgag cccaaagagc attcaatcat tgttttattt gccttctttt cacaccattg 2400
gtgagggagg gattaccacc ctggggttat gaagatggtt gaacacccca cacatagcac 2460
cggagatatg agatcaacag tttcttagcc atagagattc acagcccaga gcaggaggac 2520
gctgcacacc atgcaggatg acatggggga tgcgctcggg attggtgtga agaagcaagg 2580
actgttagag gcaggcttta tagtaacaag acggtggggc aaactctgat ttccgtgggg 2640
gaatgtcatg gtcttgcttt actaagtttt gagactggca ggtagtgaaa ctcattaggc 2700
tgagaacctt gtggaatgca gctgacccag ctgatagagg aagtagccag gtgggagcct 2760
ttcccagtgg gtgtgggaca tatctggcaa gattttgtgg cactcctggt tacagatact 2820
ggggcagcaa ataaaactga atcttgtttt cagaccttaa aaaaaaaaaa aaaaaaaaa 2879
<210>145
<211>164
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hK2 isoform 3
<400>145
Met Trp Asp Leu Val Leu Ser Ile Ala Leu Ser Val Gly Cys Thr Gly
1 5 10 15
Ala Val Pro Leu Ile Gln Ser Arg Ile Val Gly Gly Trp Glu Cys Glu
20 25 30
Lys His Ser Gln Pro Trp Gln Val Ala Val Tyr Ser His Gly Trp Ala
35 40 45
His Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala
50 55 60
His Cys Leu Lys Lys Asn Ser Gln Val Trp Leu Gly Arg His Asn Leu
65 70 75 80
Phe Glu Pro Glu Asp Thr Gly Gln Arg Val Pro Val Ser His Ser Phe
85 90 95
Pro His Pro Leu Tyr Asn Met Ser Leu Leu Lys His Gln Ser Leu Arg
100 105 110
Pro Asp Glu Asp Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser Glu
115 120 125
Pro Ala Lys Ile Thr Asp Val Val Lys Val Leu Gly Leu Pro Thr Gln
130 135 140
Glu Pro Ala Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser Ile
145 150 155 160
Glu Pro Glu Glu
<210>146
<211>1464
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein 3 transcript variant 1
<400>146
agccccaagc ttaccacctg cacccggaga gctgtgtcac catgtgggtc ccggttgtct 60
tcctcaccct gtccgtgacg tggattggtg ctgcacccct catcctgtct cggattgtgg 120
gaggctggga gtgcgagaag cattcccaac cctggcaggt gcttgtggcc tctcgtggca 180
gggcagtctg cggcggtgtt ctggtgcacc cccagtgggt cctcacagct gcccactgca 240
tcaggaacaa aagcgtgatc ttgctgggtc ggcacagcct gtttcatcct gaagacacag 300
gccaggtatt tcaggtcagc cacagcttcc cacacccgct ctacgatatg agcctcctga 360
agaatcgatt cctcaggcca ggtgatgact ccagccacga cctcatgctg ctccgcctgt 420
cagagcctgc cgagctcacg gatgctgtga aggtcatgga cctgcccacc caggagccag 480
cactggggac cacctgctac gcctcaggct ggggcagcat tgaaccagag gagttcttga 540
ccccaaagaa acttcagtgt gtggacctcc atgttatttc caatgacgtg tgtgcgcaag 600
ttcaccctca gaaggtgacc aagttcatgc tgtgtgctgg acgctggaca gggggcaaaa 660
gcacctgctc gggtgattct gggggcccac ttgtctgtaa tggtgtgctt caaggtatca 720
cgtcatgggg cagtgaacca tgtgccctgc ccgaaaggcc ttccctgtac accaaggtgg 780
tgcattaccg gaagtggatc aaggacacca tcgtggccaa cccctgagca cccctatcaa 840
ccccctattg tagtaaactt ggaaccttgg aaatgaccag gccaagactc aagcctcccc 900
agttctactg acctttgtcc ttaggtgtga ggtccagggt tgctaggaaa agaaatcagc 960
agacacaggt gtagaccaga gtgtttctta aatggtgtaa ttttgtcctc tctgtgtcct 1020
ggggaatact ggccatgcct ggagacatat cactcaattt ctctgaggac acagatagga 1080
tggggtgtct gtgttatttg tggggtacag agatgaaaga ggggtgggat ccacactgag 1140
agagtggaga gtgacatgtg ctggacactg tccatgaagc actgagcaga agctggaggc 1200
acaacgcacc agacactcac agcaaggatg gagctgaaaa cataacccac tctgtcctgg 1260
aggcactggg aagcctagag aaggctgtga gccaaggagg gagggtcttc ctttggcatg 1320
ggatggggat gaagtaagga gagggactgg accccctgga agctgattca ctatgggggg 1380
aggtgtattg aagtcctcca gacaaccctc agatttgatg atttcctagt agaactcaca 1440
gaaataaaga gctgttatac tgtg 1464
<210>147
<211>261
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein 3 isoform 1
<400>147
Met Trp Val Pro Val Val Phe Leu Thr Leu Ser Val Thr Trp Ile Gly
1 5 10 15
Ala Ala Pro Leu Ile Leu Ser Arg Ile Val Gly Gly Trp Glu Cys Glu
20 25 30
Lys His Ser Gln Pro Trp Gln Val Leu Val Ala Ser Arg Gly Arg Ala
35 40 45
Val Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala
50 55 60
His Cys Ile Arg Asn Lys Ser Val Ile Leu Leu Gly Arg His Ser Leu
65 70 75 80
Phe His Pro Glu Asp Thr Gly Gln Val Phe Gln Val Ser His Ser Phe
85 90 95
Pro His Pro Leu Tyr Asp Met Ser Leu Leu Lys Asn Arg Phe Leu Arg
100 105 110
Pro Gly Asp Asp Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser Glu
115 120 125
Pro Ala Glu Leu Thr Asp Ala Val Lys Val Met Asp Leu Pro Thr Gln
130 135 140
Glu Pro Ala Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser Ile
145 150 155 160
Glu Pro Glu Glu Phe Leu Thr Pro Lys Lys Leu Gln Cys Val Asp Leu
165 170 175
His Val Ile Ser Asn Asp Val Cys Ala Gln Val His Pro Gln Lys Val
180 185 190
Thr Lys Phe Met Leu Cys Ala Gly Arg Trp Thr Gly Gly Lys Ser Thr
195 200 205
Cys Ser Gly Asp Ser Gly Gly Pro Leu Val Cys Asn Gly Val Leu Gln
210 215 220
Gly Ile Thr Ser Trp Gly Ser Glu Pro Cys Ala Leu Pro Glu Arg Pro
225 230 235 240
Ser Leu Tyr Thr Lys Val Val His Tyr Arg Lys Trp Ile Lys Asp Thr
245 250 255
Ile Val Ala Asn Pro
260
<210>148
<211>1906
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein 3 transcript variant 3
<400>148
agccccaagc ttaccacctg cacccggaga gctgtgtcac catgtgggtc ccggttgtct 60
tcctcaccct gtccgtgacg tggattggtg ctgcacccct catcctgtct cggattgtgg 120
gaggctggga gtgcgagaag cattcccaac cctggcaggt gcttgtggcc tctcgtggca 180
gggcagtctg cggcggtgtt ctggtgcacc cccagtgggt cctcacagct gcccactgca 240
tcaggaacaa aagcgtgatc ttgctgggtc ggcacagcct gtttcatcct gaagacacag 300
gccaggtatt tcaggtcagc cacagcttcc cacacccgct ctacgatatg agcctcctga 360
agaatcgatt cctcaggcca ggtgatgact ccagccacga cctcatgctg ctccgcctgt 420
cagagcctgc cgagctcacg gatgctgtga aggtcatgga cctgcccacc caggagccag 480
cactggggac cacctgctac gcctcaggct ggggcagcat tgaaccagag gagttcttga 540
ccccaaagaa acttcagtgt gtggacctcc atgttatttc caatgacgtg tgtgcgcaag 600
ttcaccctca gaaggtgacc aagttcatgc tgtgtgctgg acgctggaca gggggcaaaa 660
gcacctgctc gtgggtcatt ctgatcaccg aactgaccat gccagccctg ccgatggtcc 720
tccatggctc cctagtgccc tggagaggag gtgtctagtc agagagtagt cctggaaggt 780
ggcctctgtg aggagccacg gggacagcat cctgcagatg gtcctggccc ttgtcccacc 840
gacctgtcta caaggactgt cctcgtggac cctcccctct gcacaggagc tggaccctga 900
agtcccttcc ccaccggcca ggactggagc ccctacccct ctgttggaat ccctgcccac 960
cttcttctgg aagtcggctc tggagacatt tctctcttct tccaaagctg ggaactgcta 1020
tctgttatct gcctgtccag gtctgaaaga taggattgcc caggcagaaa ctgggactga 1080
cctatctcac tctctccctg cttttaccct tagggtgatt ctgggggccc acttgtctgt 1140
aatggtgtgc ttcaaggtat cacgtcatgg ggcagtgaac catgtgccct gcccgaaagg 1200
ccttccctgt acaccaaggt ggtgcattac cggaagtgga tcaaggacac catcgtggcc 1260
aacccctgag cacccctatc aaccccctat tgtagtaaac ttggaacctt ggaaatgacc 1320
aggccaagac tcaagcctcc ccagttctac tgacctttgt ccttaggtgt gaggtccagg 1380
gttgctagga aaagaaatca gcagacacag gtgtagacca gagtgtttct taaatggtgt 1440
aattttgtcc tctctgtgtc ctggggaata ctggccatgc ctggagacat atcactcaat 1500
ttctctgagg acacagatag gatggggtgt ctgtgttatt tgtggggtac agagatgaaa 1560
gaggggtggg atccacactg agagagtgga gagtgacatg tgctggacac tgtccatgaa 1620
gcactgagca gaagctggag gcacaacgca ccagacactc acagcaagga tggagctgaa 1680
aacataaccc actctgtcct ggaggcactg ggaagcctag agaaggctgt gagccaagga 1740
gggagggtct tcctttggca tgggatgggg atgaagtaag gagagggact ggaccccctg 1800
gaagctgatt cactatgggg ggaggtgtat tgaagtcctc cagacaaccc tcagatttga 1860
tgatttccta gtagaactca cagaaataaa gagctgttat actgtg 1906
<210>149
<211>238
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein 3 isoform 3
<400>149
Met Trp Val Pro Val Val Phe Leu Thr Leu Ser Val Thr Trp Ile Gly
1 5 10 15
Ala Ala Pro Leu Ile Leu Ser Arg Ile Val Gly Gly Trp Glu Cys Glu
20 25 30
Lys His Ser Gln Pro Trp Gln Val Leu Val Ala Ser Arg Gly Arg Ala
35 40 45
Val Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala
50 55 60
His Cys Ile Arg Asn Lys Ser Val Ile Leu Leu Gly Arg His Ser Leu
65 70 75 80
Phe His Pro Glu Asp Thr Gly Gln Val Phe Gln Val Ser His Ser Phe
85 90 95
Pro His Pro Leu Tyr Asp Met Ser Leu Leu Lys Asn Arg Phe Leu Arg
100 105 110
Pro Gly Asp Asp Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser Glu
115 120 125
Pro Ala Glu Leu Thr Asp Ala Val Lys Val Met Asp Leu Pro Thr Gln
130 135 140
Glu Pro Ala Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser Ile
145 150 155 160
Glu Pro Glu Glu Phe Leu Thr Pro Lys Lys Leu Gln Cys Val Asp Leu
165 170 175
His Val Ile Ser Asn Asp Val Cys Ala Gln Val His Pro Gln Lys Val
180 185 190
Thr Lys Phe Met Leu Cys Ala Gly Arg Trp Thr Gly Gly Lys Ser Thr
195 200 205
Cys Ser Trp Val Ile Leu Ile Thr Glu Leu Thr Met Pro Ala Leu Pro
210 215 220
Met Val Leu His Gly Ser Leu Val Pro Trp Arg Gly Gly Val
225 230 235
<210>150
<211>1335
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein 3 transcript variant 4
<400>150
agccccaagc ttaccacctg cacccggaga gctgtgtcac catgtgggtc ccggttgtct 60
tcctcaccct gtccgtgacg tggattggtg ctgcacccct catcctgtct cggattgtgg 120
gaggctggga gtgcgagaag cattcccaac cctggcaggt gcttgtggcc tctcgtggca 180
gggcagtctg cggcggtgtt ctggtgcacc cccagtgggt cctcacagct gcccactgca 240
tcaggaagcc aggtgatgac tccagccacg acctcatgct gctccgcctg tcagagcctg 300
ccgagctcac ggatgctgtg aaggtcatgg acctgcccac ccaggagcca gcactgggga 360
ccacctgcta cgcctcaggc tggggcagca ttgaaccaga ggagttcttg accccaaaga 420
aacttcagtg tgtggacctc catgttattt ccaatgacgt gtgtgcgcaa gttcaccctc 480
agaaggtgac caagttcatg ctgtgtgctg gacgctggac agggggcaaa agcacctgct 540
cgggtgattc tgggggccca cttgtctgta atggtgtgct tcaaggtatc acgtcatggg 600
gcagtgaacc atgtgccctg cccgaaaggc cttccctgta caccaaggtg gtgcattacc 660
ggaagtggat caaggacacc atcgtggcca acccctgagc acccctatca accccctatt 720
gtagtaaact tggaaccttg gaaatgacca ggccaagact caagcctccc cagttctact 780
gacctttgtc cttaggtgtg aggtccaggg ttgctaggaa aagaaatcag cagacacagg 840
tgtagaccag agtgtttctt aaatggtgta attttgtcct ctctgtgtcc tggggaatac 900
tggccatgcc tggagacata tcactcaatt tctctgagga cacagatagg atggggtgtc 960
tgtgttattt gtggggtaca gagatgaaag aggggtggga tccacactga gagagtggag 1020
agtgacatgt gctggacact gtccatgaag cactgagcag aagctggagg cacaacgcac 1080
cagacactca cagcaaggat ggagctgaaa acataaccca ctctgtcctg gaggcactgg 1140
gaagcctaga gaaggctgtg agccaaggag ggagggtctt cctttggcat gggatgggga 1200
tgaagtaagg agagggactg gaccccctgg aagctgattc actatggggg gaggtgtatt 1260
gaagtcctcc agacaaccct cagatttgat gatttcctag tagaactcac agaaataaag 1320
agctgttata ctgtg 1335
<210>151
<211>218
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein 3 isoform 4
<400>151
Met Trp Val Pro Val Val Phe Leu Thr Leu Ser Val Thr Trp Ile Gly
1 5 10 15
Ala Ala Pro Leu Ile Leu Ser Arg Ile Val Gly Gly Trp Glu Cys Glu
20 25 30
Lys His Ser Gln Pro Trp Gln Val Leu Val Ala Ser Arg Gly Arg Ala
35 40 45
Val Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala
50 55 60
His Cys Ile Arg Lys Pro Gly Asp Asp Ser Ser His Asp Leu Met Leu
65 70 75 80
Leu Arg Leu Ser Glu Pro Ala Glu Leu Thr Asp Ala Val Lys Val Met
85 90 95
Asp Leu Pro Thr Gln Glu Pro Ala Leu Gly Thr Thr Cys Tyr Ala Ser
100 105 110
Gly Trp Gly Ser Ile Glu Pro Glu Glu Phe Leu Thr Pro Lys Lys Leu
115 120 125
Gln Cys Val Asp Leu His Val Ile Ser Asn Asp Val Cys Ala Gln Val
130 135 140
His Pro Gln Lys Val Thr Lys Phe Met Leu Cys Ala Gly Arg Trp Thr
145 150 155 160
Gly Gly Lys Ser Thr Cys Ser Gly Asp Ser Gly Gly Pro Leu ValCys
165 170 175
Asn Gly Val Leu Gln Gly Ile Thr Ser Trp Gly Ser Glu Pro Cys Ala
180 185 190
Leu Pro Glu Arg Pro Ser Leu Tyr Thr Lys Val Val His Tyr Arg Lys
195 200 205
Trp Ile Lys Asp Thr Ile Val Ala Asn Pro
210 215
<210>152
<211>1341
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein 3 transcript variant 5
<400>152
agccccaagc ttaccacctg cacccggaga gctgtgtcac catgtgggtc ccggttgtct 60
tcctcaccct gtccgtgacg tggattggtg ctgcacccct catcctgtct cggattgtgg 120
gaggctggga gtgcgagaag cattcccaac cctggcaggt gcttgtggcc tctcgtggca 180
gggcagtctg cggcggtgtt ctggtgcacc cccagtgggt cctcacagct gcccactgca 240
tcaggaacaa aagcgtgatc ttgctgggtc ggcacagcct gtttcatcct gaagacacag 300
gccaggtatt tcaggtcagc cacagcttcc cacacccgct ctacgatatg agcctcctga 360
agaatcgatt cctcaggcca ggtgatgact ccagcattga accagaggag ttcttgaccc 420
caaagaaact tcagtgtgtg gacctccatg ttatttccaa tgacgtgtgt gcgcaagttc 480
accctcagaa ggtgaccaag ttcatgctgt gtgctggacg ctggacaggg ggcaaaagca 540
cctgctcggg tgattctggg ggcccacttg tctgtaatgg tgtgcttcaa ggtatcacgt 600
catggggcag tgaaccatgt gccctgcccg aaaggccttc cctgtacacc aaggtggtgc 660
attaccggaa gtggatcaag gacaccatcg tggccaaccc ctgagcaccc ctatcaaccc 720
cctattgtag taaacttgga accttggaaa tgaccaggcc aagactcaag cctccccagt 780
tctactgacc tttgtcctta ggtgtgaggt ccagggttgc taggaaaaga aatcagcaga 840
cacaggtgta gaccagagtg tttcttaaat ggtgtaattt tgtcctctct gtgtcctggg 900
gaatactggc catgcctgga gacatatcac tcaatttctc tgaggacaca gataggatgg 960
ggtgtctgtg ttatttgtgg ggtacagaga tgaaagaggg gtgggatcca cactgagaga 1020
gtggagagtg acatgtgctg gacactgtcc atgaagcact gagcagaagc tggaggcaca 1080
acgcaccaga cactcacagc aaggatggag ctgaaaacat aacccactct gtcctggagg 1140
cactgggaag cctagagaag gctgtgagcc aaggagggag ggtcttcctt tggcatggga 1200
tggggatgaa gtaaggagag ggactggacc ccctggaagc tgattcacta tggggggagg 1260
tgtattgaag tcctccagac aaccctcaga tttgatgatt tcctagtaga actcacagaa 1320
ataaagagct gttatactgt g 1341
<210>153
<211>220
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein 3 isoform 5
<400>153
Met Trp Val Pro Val Val Phe Leu Thr Leu Ser Val Thr Trp Ile Gly
1 5 10 15
Ala Ala Pro Leu Ile Leu Ser Arg Ile Val Gly Gly Trp Glu Cys Glu
20 25 30
Lys His Ser Gln Pro Trp Gln Val Leu Val Ala Ser Arg Gly Arg Ala
35 40 45
Val Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala
50 55 60
His Cys Ile Arg Asn Lys Ser Val Ile Leu Leu Gly Arg His Ser Leu
65 70 75 80
Phe His Pro Glu Asp Thr Gly Gln Val Phe Gln Val Ser His Ser Phe
85 90 95
Pro His Pro Leu Tyr Asp Met Ser Leu Leu Lys Asn Arg Phe Leu Arg
100 105 110
Pro Gly Asp Asp Ser Ser Ile Glu Pro Glu Glu Phe Leu Thr Pro Lys
115 120 125
Lys Leu Gln Cys Val Asp Leu His Val Ile Ser Asn Asp Val Cys Ala
130 135 140
Gln Val His Pro Gln Lys Val Thr Lys Phe Met Leu Cys Ala Gly Arg
145 150 155 160
Trp Thr Gly Gly Lys Ser Thr Cys Ser Gly Asp Ser Gly Gly Pro Leu
165 170 175
Val Cys Asn Gly Val Leu Gln Gly Ile Thr Ser Trp Gly Ser Glu Pro
180 185 190
Cys Ala Leu Pro Glu Arg Pro Ser Leu Tyr Thr Lys Val Val His Tyr
195 200 205
Arg Lys Trp Ile Lys Asp Thr Ile Val Ala Asn Pro
210 215 220
<210>154
<211>555
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein 3 transcript variant 6
<400>154
agccccaagc ttaccacctg cacccggaga gctgtgtcac catgtgggtc ccggttgtct 60
tcctcaccct gtccgtgacg tggattggtg ctgcacccct catcctgtct cggattgtgg 120
gaggctggga gtgcgagaag cattcccaac cctggcaggt gcttgtggcc tctcgtggca 180
gggcagtctg cggcggtgtt ctggtgcace cccagtgggt cctcacagct gcccactgca 240
tcaggaagtg agtaggggcc tggggtctgg ggagcaggtg tctgtgtccc agaggaataa 300
cagctgggca ttttccccag gataacctct aaggccagcc ttgggactgg gggagagagg 360
gaaagttctg gttcaggtca catggggagg cagggttggg gctggaccac cctccccatg 420
gctgcctggg tctccatctg tgttcctcta tgtctctttg tgtcgctttc attatgtctc 480
ttggtaactg gcttcggttg tgtctctccg tgtgactatt ttgttctctc tctccctctc 540
ttctctgtct tcagt 555
<210>155
<211>69
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein 3 isoform 6
<400>155
Met Trp Val Pro Val Val Phe Leu Thr Leu Ser Val Thr Trp Ile Gly
1 5 10 15
Ala Ala Pro Leu Ile Leu Ser Arg Ile Val Gly Gly Trp Glu Cys Glu
20 25 30
Lys His Ser Gln Pro Trp Gln Val Leu Val Ala Ser Arg Gly Arg Ala
35 40 45
Val Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala
50 55 60
His Cys Ile Arg Lys
65
<210>156
<211>807
<212>DNA
<213>Homo sapiens
<220>
<223>Mesotrypsin
<400>156
acactctacc accatgaatc cattcctgat ccttgccttt gtgggagctg ctgttgctgt 60
cccctttgac gatgatgaca agattgttgg gggctacacc tgtgaggaga attctctccc 120
ctaccaggtg tccctgaatt ctggctccca cttctgcggt ggctccctca tcagcgaaca 180
gtgggtggta tcagcagctc actgctacaa gacccgcatc caggtgagac tgggagagca 240
caacatcaaa gtcctggagg ggaatgagca gttcatcaat gcggccaaga tcatccgcca 300
ccctaaatac aacagggaca ctctggacaa tgacatcatg ctgatcaaac tctcctcacc 360
tgccgtcatc aatgcccgcg tgtccaccat ctctctgccc accgcccctc cagctgctgg 420
cactgagtgc ctcatctccg gctggggcaa cactctgagc tttggtgctg actacccaga 480
cgagctgaag tgcctggatg ctccggtgct gacccaggct gagtgtaaag cctcctaccc 540
tggaaagatt accaacagca tgttctgtgt gggcttcctt gagggaggca aggattcctg 600
ccagcgtgac tctggtggcc ctgtggtctg caacggacag ctccaaggag ttgtctcctg 660
gggccatggc tgtgcctgga agaacaggcc tggagtctac accaaggtct acaactatgt 720
ggactggatt aaggacacca tcgctgccaa cagctaaagc ccccggtccc tctgcagtct 780
ctataccaat aaagtggccc tgctctc 807
<210>157
<211>247
<212>PRT
<213>Homo sapiens
<220>
<223>Mesotrypsin
<400>157
Met Asn Pro Phe Leu Ile Leu Ala Phe Val Gly Ala Ala Val Ala Val
1 5 10 15
Pro Phe Asp Asp Asp Asp Lys Ile Val Gly Gly Tyr Thr Cys Glu Glu
20 25 30
Asn Ser Leu Pro Tyr Gln Val Ser Leu Asn Ser Gly Ser His Phe Cys
35 40 45
Gly Gly Ser Leu Ile Ser Glu Gln Trp Val Val Ser Ala Ala His Cys
50 55 60
Tyr Lys Thr Arg Ile Gln Val Arg Leu Gly Glu His Asn Ile Lys Val
65 70 75 80
Leu Glu Gly Asn Glu Gln Phe Ile Asn Ala Ala Lys Ile Ile Arg His
85 90 95
Pro Lys Tyr Asn Arg Asp Thr Leu Asp Asn Asp Ile Met Leu Ile Lys
100 105 110
Leu Ser Ser Pro Ala Val Ile Asn Ala Arg Val Ser Thr Ile Ser Leu
115 120 125
Pro Thr Ala Pro Pro Ala Ala Gly Thr Glu Cys Leu Ile Ser Gly Trp
130 135 140
Gly Asn Thr Leu Ser Phe Gly Ala Asp Tyr Pro Asp Glu Leu Lys Cys
145 150 155 160
Leu Asp Ala Pro Val Leu Thr Gln Ala Glu Cys Lys Ala Ser Tyr Pro
165 170 175
Gly Lys Ile Thr Asn Ser Met Phe Cys Val Gly Phe Leu Glu Gly Gly
180 185 190
Lys Asp Ser Cys Gln Arg Asp Ser Gly Gly Pro Val Val Cys Asn Gly
195 200 205
Gln Leu Gln Gly Val Val Ser Trp Gly His Gly Cys Ala Trp Lys Asn
210 215 220
Arg Pro Gly Val Tyr Thr Lys Val Tyr Asn Tyr Val Asp Trp Ile Lys
225 230 235 240
Asp Thr Ile Ala Ala Asn Ser
245
<210>158
<211>1055
<212>DNA
<213>Homo sapiens
<220>
<223>pancreatic endopeptidase E form B (B)
<400>158
acctgcattt ccaccacctg ttctgggagt ggggcccagt cagccccctt tgcttacctt 60
ctgccctata ttagagcccc aggttctgtg cccttttcct atcatcgcaa aactcatgat 120
gctccggctg ctcagttccc tcctccttgt ggccgttgcc tcaggctatg gcccaccttc 180
ctctcgccct tccagccgcg ttgtcaatgg tgaggatgcg gtcccctaca gctggccctg 240
gcaggtttcc ctgcagtatg agaaaagcgg aagcttctac cacacctgtg gcggtagcct 300
catcgccccc gactgggttg tgactgccgg ccactgcatc tcgagctccc ggacctacca 360
ggtggtgttg ggcgagtacg accgtgctgt gaaggagggc cccgagcagg tgatccccat 420
caactctggg gacctctttg tgcatccact ctggaaccgc tcgtgtgtgg cctgtggcaa 480
tgacatcgcc ctcatcaagc tctcacgcag cgcccagctg ggagacgccg tccagctcgc 540
ctcactccct ccggctggtg acatccttcc caacgagaca ccctgctaca tcaccggctg 600
gggccgtctc tataccaacg ggccactccc agacaagctg caggaggccc tgctgccggt 660
ggtggactat gaacactgct ccaggtggaa ctggtggggt tcctccgtga agaagaccat 720
ggtgtgtgct ggaggggaca tccgctccgg ctgcaatggt gactctggag gacccctcaa 780
ctgccccaca gaggatggtg gctggcaggt ccatggcgtg accagctttg tttctgcctt 840
tggctgcaac acccgcagga agcccacggt gttcactcga gtctccgcct tcattgactg 900
gattgaggag accatagcaa gccactagaa ccaaggccca gctggcagtg ctgatcgatc 960
ccacatcctg aataaagaat aaagatctct cagaaaattc caaaaaaaaa aaaaaaaaaa 1020
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaa 1055
<210>159
<211>270
<212>PRT
<213>Homo sapiens
<220>
<223>pancreatic endopeptidase E form B (B)
<400>159
Met Met Leu Arg Leu Leu Ser Ser Leu Leu Leu Val Ala Val Ala Ser
1 5 10 15
Gly Tyr Gly Pro Pro Ser Ser Arg Pro Ser Ser Arg Val Val Asn Gly
20 25 30
Glu Asp Ala Val Pro Tyr Ser Trp Pro Trp Gln Val Ser Leu Gln Tyr
35 40 45
Glu Lys Ser Gly Ser Phe Tyr His Thr Cys Gly Gly Ser Leu Ile Ala
50 55 60
Pro Asp Trp Val Val Thr Ala Gly His Cys Ile Ser Ser Ser Arg Thr
65 70 75 80
Tyr Gln Val Val Leu Gly Glu Tyr Asp Arg Ala Val Lys Glu Gly Pro
85 90 95
Glu Gln Val Ile Pro Ile Asn Ser Gly Asp Leu Phe Val His Pro Leu
100 105 110
Trp Asn Arg Ser Cys Val nAla Cys Gly Asn Asp Ile Ala Leu Ile Lys
115 120 125
Leu Ser Arg Ser Ala Gln Leu Gly Asp Ala Val Gln Leu Ala Ser Leu
130 135 140
Pro Pro Ala Gly Asp Ile Leu Pro Asn Glu Thr Pro Cys Tyr Ile Thr
145 150 155 160
Gly Trp Gly Arg Leu Tyr Thr Asn Gly Pro Leu Pro Asp Lys Leu Gln
165 170 175
Glu Ala Leu Leu Pro Val Val Asp Tyr Glu His Cys Ser Arg Trp Asn
180 185 190
Trp Trp Gly Ser Ser Val nLys Lys Thr Met Val Cys Ala Gly Gly Asp
195 200 205
Ile Arg Ser Gly Cys Asn Gly Asp Ser Gly Gly Pro Leu Asn Cys Pro
210 215 220
Thr Glu Asp Gly Gly Trp Gln Val His Gly Val Thr Ser Phe Val Ser
225 230 235 240
Ala Phe Gly Cys Asn Thr Arg Arg Lys Pro Thr Val Phe Thr Arg Val
245 250 255
Ser Ala Phe Ile Asp Trp Ile Glu Glu Thr Ile Ala Ser His
260 265 270
<210>160
<211>941
<212>DNA
<213>Homo sapiens
<220>
<223>pancreatic elastase II form B (Homos sapiens)(IIB)
<400>160
cttacagaac tcccacggac acaccatgat taggaccctg ctgctgtcca ctttggtggc 60
tggagccctc agttgtgggg tctccactta cgcgcctgat atgtctagga tgcttggagg 120
tgaagaagcg aggcccaaca gctggccctg gcaggtctcc ctgcagtaca gctccaatgg 180
ccagtggtac cacacctgcg gagggtccct gatagccaac agctgggtcc tgacggctgc 240
ccactgcatc agctcctccg ggatctaccg cgtgatgctg ggccagcata acctctacgt 300
tgcagagtcc ggctcgctgg ccgtcagtgt ctctaagatt gtggtgcaca aggactggaa 360
ctccgaccag gtctccaaag ggaacgacat tgccctgctc aaactggcta accccgtctc 420
cctcaccgac aagatccagc tggcctgcct ccctcctgcc ggcaccattc tacccaacaa 480
ctacccctgc tacgtcacgg gctggggaag gctgcagacc aacggggctc tccctgatga 540
cctgaagcag ggccggttgc tggttgtgga ctatgccacc tgctccagct ctggctggtg 600
gggcagcacc gtgaagacga atatgatctg tgctgggggt gatggcgtga tatgcacctg 660
caacggagac tccggtgggc cgctgaactg tcaggcatct gacggccggt gggaggtgca 720
tggcatcggc agcctcacgt cggtccttgg ttgcaactac tactacaagc cctccatctt 780
cacgcgggtc tccaactaca acgactggat caattcggtg attgcaaata actaaccaaa 840
agaagtccct gggactgttt cagacttgga aaggtcacag aaggaaaata atattatata 900
aagtgacaac tatgcaaatc acaaaaaaaa aaaaaaaaaa a 941
<210>161
<211>269
<212>PRT
<213>Homo sapiens
<220>
<223>pancreatic elastase II form B (Homos sapiens)(IIB)
<400>161
Met Ile Arg Thr Leu Leu Leu Ser Thr Leu Val Ala Gly Ala Leu Ser
1 5 10 15
Cys Gly Val Ser Thr Tyr Ala Pro Asp Met Ser Arg Met Leu Gly Gly
20 25 30
Glu Glu Ala Arg Pro Asn Ser Trp Pro Trp Gln Val Ser Leu Gln Tyr
35 40 45
Ser Ser Asn Gly Gln Trp Tyr His Thr Cys Gly Gly Ser Leu Ile Ala
50 55 60
Asn Ser Trp Val Leu Thr Ala Ala His Cys Ile Ser Ser Ser Gly Ile
65 70 75 80
Tyr Arg Val Met Leu Gly Gln His Asn Leu Tyr Val Ala Glu Ser Gly
85 90 95
Ser Leu Ala Val Ser Val Ser Lys Ile Val Val His Lys Asp Trp Asn
100 105 110
Ser Asp Gln Val Ser Lys Gly Asn Asp Ile Ala Leu Leu Lys Leu Ala
115 120 125
Asn Pro Val Ser Leu Thr Asp Lys Ile Gln Leu Ala Cys Leu Pro Pro
130 135 140
Ala Gly Thr Ile Leu Pro Asn Asn Tyr Pro Cys Tyr Val Thr Gly Trp
145 150 155 160
Gly Arg Leu Gln Thr Asn Gly Ala Leu Pro Asp Asp Leu Lys Gln Gly
165 170 175
Arg Leu Leu Val Val Asp Tyr Ala Thr Cys Ser Ser Ser Gly Trp Trp
180 185 190
Gly Ser Thr Val Lys Thr Asn Met Ile Cys Ala Gly Gly Asp Gly Val
195 200 205
Ile Cys Thr Cys Asn Gly Asp Ser Gly Gly Pro Leu Asn Cys Gln Ala
210 215 220
Ser Asp Gly Arg Trp Glu Val His Gly Ile Gly Ser Leu Thr Ser Val
225 230 235 240
Leu Gly Cys Asn Tyr Tyr Tyr Lys Pro Ser Ile Phe Thr Arg Val Ser
245 250 255
Asn Tyr Asn Asp Trp Ile Asn Ser Val Ile Ala Asn Asn
260 265
<210>162
<211>2048
<212>DNA
<213>Homo sapiens
<220>
<223>coagulation factor XIIa
<400>162
ctattgatct ggactcctgg ataggcagct ggaccaacgg acggacgcca tgagggctct 60
gctgctcctg gggttcctgc tggtgagctt ggagtcaaca ctttcgattc caccttggga 120
agcccccaag gagcataagt acaaagctga agagcacaca gtcgttctca ctgtcaccgg 180
ggagccctgc cacttcccct tccagtacca ccggcagctg taccacaaat gtacccacaa 240
gggccggcca ggccctcagc cctggtgtgc taccaccccc aactttgatc aggaccagcg 300
atggggatac tgtttggagc ccaagaaagt gaaagaccac tgcagcaaac acagcccctg 360
ccagaaagga gggacctgtg tgaacatgcc aagcggcccc cactgtctct gtccacaaca 420
cctcactgga aaccactgcc agaaagagaa gtgctttgag cctcagcttc tccggttttt 480
ccacaagaat gagatatggt atagaactga gcaagcagct gtggccagat gccagtgcaa 540
gggtcctgat gcccactgcc agcggctggc cagccaggcc tgccgcacca acccgtgcct 600
ccatgggggt cgctgcctag aggtggaggg ccaccgcctg tgccactgcc cggtgggcta 660
caccggaccc ttctgcgacg tggacaccaa ggcaagctgc tatgatggcc gcgggctcag 720
ctaccgcggc ctggccagga ccacgctctc gggtgcgccc tgtcagccgt gggcctcgga 780
ggccacctac cggaacgtga ctgccgagca agcgcggaac tggggactgg gcggccacgc 840
cttctgccgg aacccggaca acgacatccg cccgtggtgc ttcgtgctga accgcgaccg 900
gctgagctgg gagtactgcg acctggcaca gtgccagacc ccaacccagg cggcgcctcc 960
gaccccggtg tcccctaggc ttcatgtccc actcatgccc gcgcagccgg caccgccgaa 1020
gcctcagccc acgacccgga ccccgcctca gtcccagacc ccgggagcct tgccggcgaa 1080
gcgggagcag ccgccttccc tgaccaggaa cggcccactg agctgcgggc agcggctccg 1140
caagagtctg tcttcgatga cccgcgtcgt tggcgggctg gtggcgctac gcggggcgca 1200
cccctacatc gccgcgctgt actggggcca cagtttctgc gccggcagcc tcatcgcccc 1260
ctgctgggtg ctgacggccg ctcactgcct gcaggaccgg cccgcacccg aggatctgac 1320
ggtggtgctc ggccaggaac gccgtaacca cagctgtgag ccgtgccaga cgttggccgt 1380
gcgctcctac cgcttgcacg aggccttctc gcccgtcagc taccagcacg acctggctct 1440
gttgcgcctt caggaggatg cggacggcag ctgcgcgctc ctgtcgcctt acgttcagcc 1500
ggtgtgcctg ccaagcggcg ccgcgcgacc ctccgagacc acgctctgcc aggtggccgg 1560
ctggggccac cagttcgagg gggcggagga atatgccagc ttcctgcagg aggcgcaggt 1620
accgttcctc tccctggagc gctgctcagc cccggacgtg cacggatcct ccatcctccc 1680
cggcatgctc tgcgcagggt tcctcgaggg cggcaccgat gcgtgccagg gtgattccgg 1740
aggcccgctg gtgtgtgagg accaagctgc agagcgccgg ctcaccctgc aaggcatcat 1800
cagctgggga tcgggctgtg gtgaccgcaa caagccaggc gtctacaccg atgtggccta 1860
ctacctggcc tggatccggg agcacaccgt ttcctgattg ctcagggact catctttccc 1920
tccttggtga ttccgcagtg agagagtggc tggggcatgg aaggcaagat tgtgtcccat 1980
tcccccagtg cggccagctc cgcgccagga tggcgaggaa ctcaataaag tgctttgaaa 2040
atgctgag 2048
<210>163
<211>615
<212>PRT
<213>Homo sapiens
<220>
<223>coagulation factor XIIa
<400>163
Met Arg Ala Leu Leu Leu Leu Gly Phe Leu Leu Val Ser Leu Glu Ser
1 5 10 15
Thr Leu Ser Ile Pro Pro Trp Glu Ala Pro Lys Glu His Lys Tyr Lys
20 25 30
Ala Glu Glu His Thr Val Val Leu Thr Val Thr Gly Glu Pro Cys His
35 40 45
Phe Pro Phe Gln Tyr His Arg Gln Leu Tyr His Lys Cys Thr His Lys
50 55 60
Gly Arg Pro Gly Pro Gln Pro Trp Cys Ala Thr Thr Pro Asn Phe Asp
65 70 75 80
Gln Asp Gln Arg Trp Gly Tyr Cys Leu Glu Pro Lys Lys Val Lys Asp
85 90 95
His Cys Ser Lys His Ser Pro Cys Gln Lys Gly Gly Thr Cys Val Asn
100 105 110
Met Pro Ser Gly Pro His Cys Leu Cys Pro Gln His Leu Thr Gly Asn
115 120 125
His Cys Gln Lys Glu Lys Cys Phe Glu Pro Gln Leu Leu Arg Phe Phe
130 135 140
His Lys Asn Glu Ile Trp Tyr Arg Thr Glu Gln Ala Ala Val Ala Arg
145 150 155 160
Cys Gln Cys Lys Gly Pro Asp Ala His Cys Gln Arg Leu Ala Ser Gln
165 170 175
Ala Cys Arg Thr Asn Pro Cys Leu His Gly Gly Arg Cys Leu Glu Val
180 185 190
Glu Gly His Arg Leu Cys His Cys Pro Val Gly Tyr Thr Gly Pro Phe
195 200 205
Cys Asp Val Asp Thr Lys Ala Ser Cys Tyr Asp Gly Arg Gly Leu Ser
210 215 220
Tyr Arg Gly Leu Ala Arg Thr Thr Leu Ser Gly Ala Pro Cys Gln Pro
225 230 235 240
Trp Ala Ser Glu Ala Thr Tyr Arg Asn Val Thr Ala Glu Gln Ala Arg
245 250 255
Asn Trp Gly Leu Gly Gly His Ala Phe Cys Arg Asn Pro Asp Asn Asp
260 265 270
Ile Arg Pro Trp Cys Phe Val Leu Asn Arg Asp Arg Leu Ser Trp Glu
275 280 285
Tyr Cys Asp Leu Ala Gln Cys Gln Thr Pro Thr Gln Ala Ala Pro Pro
290 295 300
Thr Pro Val Ser Pro Arg Leu His Val Pro Leu Met Pro Ala Gln Pro
305 310 315 320
Ala Pro Pro Lys Pro Gln Pro Thr Thr Arg Thr Pro Pro Gln Ser Gln
325 330 335
Thr Pro Gly Ala Leu Pro Ala Lys Arg Glu Gln Pro Pro Ser Leu Thr
340 345 350
Arg Asn Gly Pro Leu Ser Cys Gly Gln Arg Leu Arg Lys Ser Leu Ser
355 360 365
Ser Met Thr Arg Val Val Gly Gly Leu Val Ala Leu Arg Gly Ala His
370 375 380
Pro Tyr Ile Ala Ala Leu Tyr Trp Gly His Ser Phe Cys Ala Gly Ser
385 390 395 400
Leu Ile Ala Pro Cys Trp Val Leu Thr Ala Ala His Cys Leu Gln Asp
405 410 415
Arg Pro Ala Pro Glu Asp Leu Thr Val Val Leu Gly Gln Glu Arg Arg
420 425 430
Asn His Ser Cys Glu Pro Cys Gln Thr Leu Ala Val Arg Ser Tyr Arg
435 440 445
Leu His Glu Ala Phe Ser Pro Val Ser Tyr Gln His Asp Leu Ala Leu
450 455 460
Leu Arg Leu Gln Glu Asp Ala Asp Gly Ser Cys Ala Leu Leu Ser Pro
465 470 475 480
Tyr Val Gln Pro Val Cys Leu Pro Ser Gly Ala Ala Arg Pro Ser Glu
485 490 495
Thr Thr Leu Cys Gln Val Ala Gly Trp Gly His Gln Phe Glu Gly Ala
500 505 510
Glu Glu Tyr Ala Ser Phe Leu Gln Glu Ala Gln Val Pro Phe Leu Ser
515 520 525
Leu Glu Arg Cys Ser Ala Pro Asp Val His Gly Ser Ser Ile Leu Pro
530 535 540
Gly Met Leu Cys Ala Gly Phe Leu Glu Gly Gly Thr Asp Ala Cys Gln
545 550 555 560
Gly Asp Ser Gly Gly Pro Leu Val Cys Glu Asp Gln Ala Ala Glu Arg
565 570 575
Arg Leu Thr Leu Gln Gly Ile Ile Ser Trp Gly Ser Gly Cys Gly Asp
580 585 590
Arg Asn Lys Pro Gly Val Tyr Thr Asp Val Ala Tyr Tyr Leu Ala Trp
595 600 605
Ile Arg Glu His Thr Val Ser
610 615
<210>164
<211>2245
<212>DNA
<213>Homo sapiens
<220>
<223>plasma kallikrein
<400>164
agaacagctt gaagaccgtt catttttaag tgacaagaga ctcacctcca agaagcaatt 60
gtgttttcag aatgatttta ttcaagcaag caacttattt catttccttg tttgctacag 120
tttcctgtgg atgtctgact caactctatg aaaacgcctt cttcagaggt ggggatgtag 180
cttccatgta caccccaaat gcccaatact gccagatgag gtgcacattc cacccaaggt 240
gtttgctatt cagttttctt ccagcaagtt caatcaatga catggagaaa aggtttggtt 300
gcttcttgaa agatagtgtt acaggaaccc tgccaaaagt acatcgaaca ggtgcagttt 360
ctggacattc cttgaagcaa tgtggtcatc aaataagtgc ttgccatcga gacatttata 420
aaggagttga tatgagagga gtcaatttta atgtgtctaa ggttagcagt gttgaagaat 480
gccaaaaaag gtgcaccaat aacattcgct gccagttttt ttcatatgcc acgcaaacat 540
ttcacaaggc agagtaccgg aacaattgcc tattaaagta cagtcccgga ggaacaccta 600
ccgctataaa ggtgctgagt aacgtggaat ctggattctc actgaagccc tgtgcccttt 660
cagaaattgg ttgccacatg aacatcttcc agcatcttgc gttctcagat gtggatgttg 720
ccagggttct cactccagat gcttttgtgt gtcggaccat ctgcacctat caccccaact 780
gcctcttctt tacattctat acaaatgtat ggaaaatcga gtcacaaaga aatgtttgtc 840
ttcttaaaac atctgaaagt ggcacaccaa gttcctctac tcctcaagaa aacaccatat 900
ctggatatag ccttttaacc tgcaaaagaa ctttacctga accctgccat tctaaaattt 960
acccgggagt tgactttgga ggagaagaat tgaatgtgac ttttgttaaa ggagtgaatg 1020
tttgccaaga gacttgcaca aagatgattc gctgtcagtt tttcacttat tctttactcc 1080
cagaagactg taaggaagag aagtgtaagt gtttcttaag attatctatg gatggttctc 1140
caactaggat tgcgtatggg acacaaggga gctctggtta ctctttgaga ttgtgtaaca 1200
ctggggacaa ctctgtctgc acaacaaaaa caagcacacg cattgttgga ggaacaaact 1260
cttcttgggg agagtggccc tggcaggtga gcctgcaggt gaagctgaca gctcagaggc 1320
acctgtgtgg agggtcactc ataggacacc agtgggtcct cactgctgcc cactgctttg 1380
atgggcttcc cctgcaggat gtttggcgca tctatagtgg cattttaaat ctgtcagaca 1440
ttacaaaaga tacacctttc tcacaaataa aagagattat tattcaccaa aactataaag 1500
tctcagaagg gaatcatgat atcgccttga taaaactcca ggctcctttg aattacactg 1560
aattccaaaa accaatatgc ctaccttcca aaggtgacac aagcacaatt tataccaact 1620
gttgggtaac cggatggggc ttctcgaagg agaaaggtga aatccaaaat attctacaaa 1680
aggtaaatat tcctttggta acaaatgaag aatgccagaa aagatatcaa gattataaaa 1740
taacccaacg gatggtctgt gctggctata aagaaggggg aaaagatgct tgtaagggag 1800
attcaggtgg tcccttagtt tgcaaacaca acggaatgtg gcgtttggtg ggcatcacaa 1860
gctggggtga aggctgtgcc cgcagggagc aacctggtgt ctacaccaaa gtcgctgagt 1920
acatggactg gattttagag aaaacacaga gcagtgatgg aaaagctcag atgcagtcac 1980
cagcatgaga agcagtccag agtctaggca atttttacaa cctgagttca agtcaaattc 2040
tgagcctggg gggtcctcat ctgcaaagca tggagagtgg catcttcttt gcatcctaag 2100
gacgaaagac acagtgcact cagagctgct gaggacaatg tctgctgaag cccgctttca 2160
gcacgccgta accaggggct gacaatgcga ggtcgcaact gagatctcca tgactgtgtg 2220
ttgtgaaata aaatggtgaa agatc 2245
<210>165
<211>638
<212>PRT
<213>Homo sapiens
<220>
<223>plasma kallikrein
<400>165
Met Ile Leu Phe Lys Gln Ala Thr Tyr Phe Ile Ser Leu Phe Ala Thr
1 5 10 15
Val Ser Cys Gly Cys Leu Thr Gln Leu Tyr Glu Asn Ala Phe Phe Arg
20 25 30
Gly Gly Asp Val Ala Ser Met Tyr Thr Pro Asn Ala Gln Tyr Cys Gln
35 40 45
Met Arg Cys Thr Phe His Pro Arg Cys Leu Leu Phe Ser Phe Leu Pro
50 55 60
Ala Ser Ser Ile Asn Asp Met Glu Lys Arg Phe Gly Cys Phe Leu Lys
65 70 75 80
Asp Ser Val Thr Gly Thr Leu Pro Lys Val His Arg Thr Gly Ala Val
85 90 95
Ser Gly His Ser Leu Lys Gln Cys Gly His Gln Ile Ser Ala Cys His
100 105 110
Arg Asp Ile Tyr Lys Gly Val Asp Met Arg Gly Val Asn Phe Asn Val
115 120 125
Ser Lys Val Ser Ser Val Glu Glu Cys Gln Lys Arg Cys Thr Asn Asn
130 135 140
Ile Arg Cys Gln Phe Phe Ser Tyr Ala Thr Gln Thr Phe His Lys Ala
145 150 155 160
Glu Tyr Arg Asn Asn Cys Leu Leu Lys Tyr Ser Pro Gly Gly Thr Pro
165 170 175
Thr Ala Ile Lys Val Leu Ser Asn Val Glu Ser Gly Phe Ser Leu Lys
180 185 190
Pro Cys Ala Leu Ser Glu Ile Gly Cys His Met Asn Ile Phe Gln His
195 200 205
Leu Ala Phe Ser Asp Val Asp Val Ala Arg Val Leu Thr Pro Asp Ala
210 215 220
Phe Val Cys Arg Thr Ile Cys Thr Tyr His Pro Asn Cys Leu Phe Phe
225 230 235 240
Thr Phe Tyr Thr Asn Val Trp Lys Ile Glu Ser Gln Arg Asn Val Cys
245 250 255
Leu Leu Lys Thr Ser Glu Ser Gly Thr Pro Ser Ser Ser Thr Pro Gln
260 265 270
Glu Asn Thr Ile Ser Gly Tyr Ser Leu Leu Thr Cys Lys Arg Thr Leu
275 280 285
Pro Glu Pro Cys His Ser Lys Ile Tyr Pro Gly Val Asp Phe Gly Gly
290 295 300
Glu Glu Leu Asn Val Thr Phe Val Lys Gly Val Asn Val Cys Gln Glu
305 310 315 320
Thr Cys Thr Lys Met Ile Arg Cys Gln Phe Phe Thr Tyr Ser Leu Leu
325 330 335
Pro Glu Asp Cys Lys Glu Glu Lys Cys Lys Cys Phe Leu Ara Leu Ser
340 345 350
Met Asp Gly Ser Pro Thr Arg Ile Ala Tyr Gly Thr Gln Gly Ser Ser
355 360 365
Gly Tyr Ser Leu Arg Leu Cys Asn Thr Gly Asp Asn Ser Val Cys Thr
370 375 380
Thr Lys Thr Ser Thr Arg Ile Val Gly Gly Thr Asn Ser Ser Trp Gly
385 390 395 400
Glu Trp Pro Trp Gln Val Ser Leu Gln Val Lys Leu Thr Ala Gln Arg
405 410 415
His Leu Cys Gly Gly Ser Leu Ile Gly His Gln Trp Val Leu Thr Ala
420 425 430
Ala His Cys Phe Asp Gly Leu Pro Leu Gln Asp Val Trp Arg Ile Tyr
435 440 445
Ser Gly Ile Leu Asn Leu Ser Asp Ile Thr Lys Asp Thr Pro Phe Ser
450 455 460
Gln Ile Lys Glu Ile Ile Ile His Gln Asn Tyr Lys Val Ser Glu Gly
465 470 475 480
Asn His Asp Ile Ala Leu Ile Lys Leu Gln Ala Pro Leu Asn Tyr Thr
485 490 495
Glu Phe Gln Lys Pro Ile Cys Leu Pro Ser Lys Gly Asp Thr Ser Thr
500 505 510
Ile Tyr Thr Asn Cys Trp Val Thr Gly Trp Gly Phe Ser Lys Glu Lys
515 520 525
Gly Glu Ile Gln Asn Ile Leu Gln Lys Val Asn Ile Pro Leu Val Thr
530 535 540
Asn Glu Glu Cys Gln Lys Arg Tyr Gln Asp Tyr Lys Ile Thr Gln Arg
545 550 555 560
Met Val Cys Ala Gly Tyr Lys Glu Gly Gly Lys Asp Ala Cys Lys Gly
565 570 575
Asp Ser Gly Gly Pro Leu Val Cys Lys His Asn Gly Met Trp Arg Leu
580 585 590
Val Gly Ile Thr Ser Trp Gly Glu Gly Cys Ala Arg Arg Glu Gln Pro
595 600 605
Gly Val Tyr Thr Lys Val Ala Glu Tyr Met Asp Trp Ile Leu Glu Lys
610 615 620
Thr Gln Ser Ser Asp Gly Lys Ala Gln Met Gln Ser Pro Ala
625 630 635
<210>166
<211>2217
<212>DNA
<213>Homo sapiens
<220>
<223>coagulation factor XIa transcript variant 1
<400>166
tgctgtactg agagagaatg ttacataaca ttgagaacta gtacaagtaa atattaaagt 60
gaagtgacca tttcctacac aagctcattc agaggaggat gaagaccatt ttggaggaag 120
aaaagcaccc ttattaagaa ttgcagcaag taagccaaca aggtcttttc aggatgattt 180
tcttatatca agtggtacat ttcattttat ttacttcagt ttctggtgaa tgtgtgactc 240
agttgttgaa ggacacctgc tttgaaggag gggacattac tacggtcttc acaccaagcg 300
ccaagtactg ccaggtagtc tgcacttacc acccaagatg tttactcttc actttcacgg 360
cggaatcacc atctgaggat cccacccgat ggtttacttg tgtcctgaaa gacagtgtta 420
cagaaacact gccaagagtg aataggacag cagcgatttc tgggtattct ttcaagcaat 480
gctcacacca aataagcgct tgcaacaaag acatttatgt ggacctagac atgaagggca 540
taaactataa cagctcagtt gccaagagtg ctcaagaatg ccaagaaaga tgcacggatg 600
acgtccactg ccactttttc acgtacgcca caaggcagtt tcccagcctg gagcatcgta 660
acatttgtct actgaagcac acccaaacag ggacaccaac cagaataacg aagctcgata 720
aagtggtgtc tggattttca ctgaaatcct gtgcactttc taatctggct tgtattaggg 780
acattttccc taatacggtg tttgcagaca gcaacatcga cagtgtcatg gctcccgatg 840
cttttgtctg tggccgaatc tgcactcatc atcccggttg cttgtttttt accttctttt 900
cccaggaatg gcccaaagaa tctcaaagaa atctttgtct ccttaaaaca tctgagagtg 960
gattgcccag tacacgcatt aaaaagagca aagctctttc tggtttcagt ctacaaagct 1020
gcaggcacag catcccagtg ttctgccatt cttcatttta ccatgacact gatttcttgg 1080
gagaagaact ggatattgtt gctgcaaaaa gtcacgaggc ctgccagaaa ctgtgcacca 1140
atgccgtccg ctgccagttt tttacctata ccccagccca agcatcctgc aacgaaggga 1200
agggcaagtg ttacttaaag ctttcttcaa acggatctcc aactaaaata cttcacggga 1260
gaggaggcat ctctggatac acattaaggt tgtgtaaaat ggataatgag tgtaccacca 1320
aaatcaagcc caggatcgtt ggaggaactg cgtctgttcg tggtgagtgg ccgtggcagg 1380
tgaccctgca cacaacctca cccactcaga gacacctgtg tggaggctcc atcattggaa 1440
accagtggat attaacagcc gctcactgtt tctatggggt agagtcacct aagattttgc 1500
gtgtctacag tggcatttta aatcaatctg aaataaaaga ggacacatct ttctttgggg 1560
ttcaagaaat aataatccat gatcagtata aaatggcaga aagcgggtat gatattgcct 1620
tgttgaaact ggaaaccaca gtgaattaca cagattctca acgacccata tgcctgcctt 1680
ccaaaggaga tagaaatgta atatacactg attgctgggt gactggatgg gggtacagaa 1740
aactaagaga caaaatacaa aatactctcc agaaagccaa gataccctta gtgaccaacg 1800
aagagtgcca gaagagatac agaggacata aaataaccca taagatgatc tgtgccggct 1860
acagggaagg agggaaggac gcttgcaagg gagattcggg aggccctctg tcctgcaaac 1920
acaatgaggt ctggcatctg gtaggcatca cgagctgggg cgaaggctgt gctcaaaggg 1980
agcggccagg tgtttacacc aacgtggtcg agtacgtgga ctggattctg gagaaaactc 2040
aagcagtgtg aatgggttcc caggggccat tggagtccct gaaggaccca ggatttgctg 2100
ggagagggtg ttgagttcac tgtgccagca tgcttcctcc acagtaacac gctgaagggg 2160
cttggtgttt gtaagaaaat gctagaagaa aacaaactgt cacaagttgt tatgtcc 2217
<210>167
<211>625
<212>PRT
<213>Homo sapiens
<220>
<223>coagulation factor XIa isoform 1
<400>167
Met Ile Phe Leu Tyr Gln Val Val His Phe Ile Leu Phe Thr Ser Val
1 5 10 15
Ser Gly Glu Cys Val Thr Gln Leu Leu Lys Asp Thr Cys Phe Glu Gly
20 25 30
Gly Asp Ile Thr Thr Val Phe Thr Pro Ser Ala Lys Tyr Cys Gln Val
35 40 45
Val Cys Thr Tyr His Pro Arg Cys Leu Leu Phe Thr Phe Thr Ala Glu
50 55 60
Ser Pro Ser Glu Asp Pro Thr Arg Trp Phe Thr Cys Val Leu Lys Asp
65 70 75 80
Ser ValThr Glu Thr Leu Pro Arg Val Asn Arg Thr Ala Ala Ile Ser
85 90 95
Gly Tyr Ser Phe Lys Gln Cys Ser His Gln Ile Ser Ala Cys Asn Lys
100 105 110
Asp Ile Tyr Val Asp Leu Asp Met Lys Gly Ile Asn Tyr Asn Ser Ser
115 120 125
Val Ala Lys Ser Ala Gln Glu Cys Gln Glu Arg Cys Thr Asp Asp Val
130 135 140
His Cys His Phe Phe Thr Tyr Ala Thr Arg Gln Phe Pro Ser Leu Glu
145 150 155 160
His Arg Asn Ile Cys Leu Leu Lys His Thr Gln Thr Gly Thr Pro Thr
165 170 175
Arg Ile Thr Lys Leu Asp Lys Val Val Ser Gly Phe Ser Leu Lys Ser
180 185 190
Cys Ala Leu Ser Asn Leu Ala Cys Ile Arg Asp Ile Phe Pro Asn Thr
195 200 205
Val Phe Ala Asp Ser Asn Ile Asp Ser Val Met Ala Pro Asp Ala Phe
210 215 220
Val Cys Gly Arg Ile Cys Thr His His Pro Gly Cys Leu Phe Phe Thr
225 230 235 240
Phe Phe Ser Gln Glu Trp Pro Lys Glu Ser Gln Arg Asn Leu Cys Leu
245 250 255
Leu Lys Thr Ser Glu Ser Gly Leu Pro Ser Thr Arg Ile Lys Lys Ser
260 265 270
Lys Ala Leu Ser Gly Phe Ser Leu Gln Ser Cys Arg His Ser Ile Pro
275 280 285
Val Phe Cys His Ser Ser Phe Tyr His Asp Thr Asp Phe Leu Gly Glu
290 295 300
Glu Leu Asp Ile Val Ala Ala Lys Ser His Glu Ala Cys Gln Lys Leu
305 310 315 320
Cys Thr Asn Ala Val Arg Cys Gln Phe Phe Thr Tyr Thr Pro Ala Gln
325 330 335
Ala Ser Cys Asn Glu Gly Lys Gly Lys Cys Tyr Leu Lys Leu Ser Ser
340 345 350
Asn Gly Ser Pro Thr Lys Ile Leu His Gly Arg Gly Gly Ile Ser Gly
355 360 365
Tyr Thr Leu Arg Leu Cys Lys Met Asp Asn Glu Cys Thr Thr Lys Ile
370 375 380
Lys Pro Arg Ile Val Gly Gly Thr Ala Ser Val Arg Gly Glu Trp Pro
385 390 395 400
Trp Gln Val Thr Leu His Thr Thr Ser Pro Thr Gln Arg His Leu Cys
405 410 415
Gly Gly Ser Ile Ile Gly Asn Gln Trp Ile Leu Thr Ala Ala His Cys
420 425 430
Phe Tyr Gly Val Glu Ser Pro Lys Ile Leu Arg Val Tyr Ser Gly Ile
435 440 445
Leu Asn Gln Ser Glu Ile Lys Glu Asp Thr Ser Phe Phe Gly Val Gln
450 455 460
Glu Ile Ile Ile His Asp Gln Tyr Lys Met Ala Glu Ser Gly Tyr Asp
465 470 475 480
Ile Ala Leu Leu Lys Leu Glu Thr Thr Val Asn Tyr Thr Asp Ser Gln
485 490 495
Arg Pro Ile Cys Leu Pro Ser Lys Gly Asp Arg Asn Val Ile Tyr Thr
500 505 510
Asp Cys Trp Val Thr Gly Trp Gly Tyr Arg Lys Leu Arg Asp Lys Ile
515 520 525
Gln Asn Thr Leu Gln Lys Ala Lys Ile Pro Leu Val Thr Asn Glu Glu
530 535 540
Cys Gln Lys Arg Tyr Arg Gly His Lys Ile Thr His Lys Met Ile Cys
545 550 555 560
Ala Gly Tyr Arg Glu Gly Gly Lys Asp Ala Cys Lys Gly Asp Ser Gly
565 570 575
Gly Pro Leu Ser Cys Lys His Asn Glu Val Trp His Leu Val Gly Ile
580 585 590
Thr Ser Trp Gly Glu Gly Cys Ala Gln Arg Glu Arg Pro Gly Val Tyr
595 600 605
Thr Asn Val Val Glu Tyr Val Asp Trp Ile Leu Glu Lys Thr Gln Ala
610 615 620
Val
625
<210>168
<211>2055
<212>DNA
<213>Homo sapiens
<220>
<223>coagulation factor XIa transcript variant 2
<400>168
tgctgtactg agagagaatg ttacataaca ttgagaacta gtacaagtaa atattaaagt 60
gaagtgacca tttcctacac aagctcattc agaggaggat gaagaccatt ttggaggaag 120
aaaagcaccc ttattaagaa ttgcagcaag taagccaaca aggtcttttc aggatgattt 180
tcttatatca agtggtacat ttcattttat ttacttcagt ttctggtgaa tgtgtgactc 240
agttgttgaa ggacacctgc tttgaaggag gggacattac tacggtcttc acaccaagcg 300
ccaagtactg ccaggtagtc tgcacttacc acccaagatg tttactcttc actttcacgg 360
cggaatcacc atctgaggat cccacccgat ggtttacttg tgtcctgaaa gacagtgtta 420
cagaaacact gccaagagtg aataggacag cagcgatttc tgggtattct ttcaagcaat 480
gctcacacca aataagtaac atttgtctac tgaagcacac ccaaacaggg acaccaacca 540
gaataacgaa gctcgataaa gtggtgtctg gattttcact gaaatcctgt gcactttcta 600
atctggcttg tattagggac attttcccta atacggtgtt tgcagacagc aacatcgaca 660
gtgtcatggc tcccgatgct tttgtctgtg gccgaatctg cactcatcat cccggttgct 720
tgttttttac cttcttttcc caggaatggc ccaaagaatc tcaaagaaat ctttgtctcc 780
ttaaaacatc tgagagtgga ttgcccagta cacgcattaa aaagagcaaa gctctttctg 840
gtttcagtct acaaagctgc aggcacagca tcccagtgtt ctgccattct tcattttacc 900
atgacactga tttcttggga gaagaactgg atattgttgc tgcaaaaagt cacgaggcct 960
gccagaaact gtgcaccaat gccgtccgct gccagttttt tacctatacc ccagcccaag 1020
catcctgcaa cgaagggaag ggcaagtgtt acttaaagct ttcttcaaac ggatctccaa 1080
ctaaaatact tcacgggaga ggaggcatct ctggatacac attaaggttg tgtaaaatgg 1140
ataatgagtg taccaccaaa atcaagccca ggatcgttgg aggaactgcg tctgttcgtg 1200
gtgagtggcc gtggcaggtg accctgcaca caacctcacc cactcagaga cacctgtgtg 1260
gaggctccat cattggaaac cagtggatat taacagccgc tcactgtttc tatggggtag 1320
agtcacctaa gattttgcgt gtctacagtg gcattttaaa tcaatctgaa ataaaagagg 1380
acacatcttt ctttggggtt caagaaataa taatccatga tcagtataaa atggcagaaa 1440
gcgggtatga tattgccttg ttgaaactgg aaaccacagt gaattacaca gattctcaac 1500
gacccatatg cctgccttcc aaaggagata gaaatgtaat atacactgat tgctgggtga 1560
ctggatgggg gtacagaaaa ctaagagaca aaatacaaaa tactctccag aaagccaaga 1620
tacccttagt gaccaacgaa gagtgccaga agagatacag aggacataaa ataacccata 1680
agatgatctg tgccggctac agggaaggag ggaaggacgc ttgcaaggga gattcgggag 1740
gccctctgtc ctgcaaacac aatgaggtct ggcatctggt aggcatcacg agctggggcg 1800
aaggctgtgc tcaaagggag cggccaggtg tttacaccaa cgtggtcgag tacgtggact 1860
ggattctgga gaaaactcaa gcagtgtgaa tgggttccca ggggccattg gagtccctga 1920
aggacccagg atttgctggg agagggtgtt gagttcactg tgccagcatg cttcctccac 1980
agtaacacgc tgaaggggct tggtgtttgt aagaaaatgc tagaagaaaa caaactgtca 2040
caagttgtta tgtcc 2055
<210>169
<211>571
<212>PRT
<213>Homo sapiens
<220>
<223>coagulation factor XIa isoform 2
<400>169
Met Ile Phe Leu Tyr Gln Val Val His Phe Ile Leu Phe Thr Ser Val
1 5 10 15
Ser Gly Glu Cys Val Thr Gln Leu Leu Lys Asp Thr Cys Phe Glu Gly
20 25 30
Gly Asp Ile Thr Thr Val Phe Thr Pro Ser Ala Lys Tyr Cys Gln Val
35 40 45
Val Cys Thr Tyr His Pro Arg Cys Leu Leu Phe Thr Phe Thr Ala Glu
50 55 60
Ser Pro Ser Glu Asp Pro Thr Arg Trp Phe Thr Cys Val Leu Lys Asp
65 70 75 80
Ser Val Thr Glu Thr Leu Pro Arg Val Asn Arg Thr Ala Ala Ile Ser
85 90 95
Gly Tyr Ser Phe Lys Gln Cys Ser His Gln Ile Ser Asn Ile Cys Leu
100 105 110
Leu Lys His Thr Gln Thr Gly Thr Pro Thr Arg Ile Thr Lys Leu Asp
115 120 125
Lys Val Val Ser Gly Phe Ser Leu Lys Ser Cys Ala Leu Ser Asn Leu
130 135 140
Ala Cys Ile Arg Asp Ile Phe Pro Asn Thr Val Phe Ala Asp Ser Asn
145 150 155 160
Ile Asp Ser Val Met Ala Pro Asp Ala Phe Val Cys Gly Arg Ile Cys
165 170 175
Thr His His Pro Gly Cys Leu Phe Phe Thr Phe Phe Ser Gln Glu Trp
180 185 190
Pro Lys Glu Ser Gln Arg Asn Leu Cys Leu Leu Lys Thr Ser Glu Ser
195 200 205
Gly Leu Pro Ser Thr Arg Ile Lys Lys Ser Lys Ala Leu Ser Gly Phe
210 215 220
Ser Leu Gln Ser Cys Arg His Ser Ile Pro Val Phe Cys His Ser Ser
225 230 235 240
Phe Tyr His Asp Thr Asp Phe Leu Gly Glu Glu Leu Asp Ile ValAla
245 250 255
Ala Lys Ser His Glu Ala Cys Gln Lys Leu Cys Thr Asn Ala Val Arg
260 265 270
Cys Gln Phe Phe Thr Tyr Thr Pro Ala Gln Ala Ser Cys Asn Glu Gly
275 280 285
Lys Gly Lys Cys Tyr Leu Lys Leu Ser Ser Asn Gly Ser Pro Thr Lys
290 295 300
Ile Leu His Gly Arg Gly Gly Ile Ser Gly Tyr Thr Leu Arg Leu Cys
305 310 315 320
Lys Met Asp Asn Glu Cys Thr Thr Lys Ile Lys Pro Arg Ile Val Gly
325 330 335
Gly Thr Ala Ser Val Arg Gly Glu Trp Pro Trp Gln Val Thr Leu His
340 345 350
Thr Thr Ser Pro Thr Gln Arg His Leu Cys Gly Gly Ser Ile Ile Gly
355 360 365
Asn Gln Trp Ile Leu Thr Ala Ala His Cys Phe Tyr Gly Val Glu Ser
370 375 380
Pro Lys Ile Leu Arg Val Tyr Ser Gly Ile Leu Asn Gln Ser Glu Ile
385 390 395 400
Lys Glu Asp Thr Ser Phe Phe Gly Val Gln Glu Ile Ile Ile His Asp
405 410 415
Gln Tyr Lys Met Ala Glu Ser Gly Tyr Asp Ile Ala Leu Leu Lys Leu
420 425 430
Glu Thr Thr Val Asn Tyr Thr Asp Ser Gln Arg Pro Ile Cys Leu Pro
435 440 445
Ser Lys Gly Asp Arg Asn Val Ile Tyr Thr Asp Cys Trp Val Thr Gly
450 455 460
Trp Gly Tyr Arg Lys Leu Arg Asp Lys Ile Gln Asn Thr Leu Gln Lys
465 470 475 480
Ala Lys Ile Pro Leu Val Thr Asn Glu Glu Cys Gln Lys Arg Tyr Arg
485 490 495
Gly His Lys Ile Thr His Lys Met Ile Cys Ala Gly Tyr Arg Glu Gly
500 505 510
Gly Lys Asp Ala Cys Lys Gly Asp Ser Gly Gly Pro Leu Ser Cys Lys
515 520 525
His Asn Glu Val Trp His Leu Val Gly Ile Thr Ser Trp Gly Glu Gly
530 535 540
Cys Ala Gln Arg Glu Arg Pro Gly Val Tyr Thr Asn Val Val Glu Tyr
545 550 555 560
Val Asp Trp Ile Leu Glu Lys Thr Gln Ala Val
565 570
<210>170
<211>2804
<212>DNA
<213>Homo sapiens
<220>
<223>coagulation factor IXa
<400>170
accactttca caatctgcta gcaaaggtta tgcagcgcgt gaacatgatc atggcagaat 60
caccaggcct catcaccatc tgccttttag gatatctact cagtgctgaa tgtacagttt 120
ttcttgatca tgaaaacgcc aacaaaattc tgaatcggcc aaagaggtat aattcaggta 180
aattggaaga gtttgttcaa gggaaccttg agagagaatg tatggaagaa aagtgtagtt 240
ttgaagaagc acgagaagtt tttgaaaaca ctgaaagaac aactgaattt tggaagcagt 300
atgttgatgg agatcagtgt gagtccaatc catgtttaaa tggcggcagt tgcaaggatg 360
acattaattc ctatgaatgt tggtgtccct ttggatttga aggaaagaac tgtgaattag 420
atgtaacatg taacattaag aatggcagat gcgagcagtt ttgtaaaaat agtgctgata 480
acaaggtggt ttgctcctgt actgagggat atcgacttgc agaaaaccag aagtcctgtg 540
aaccagcagt gccatttcca tgtggaagag tttctgtttc acaaacttct aagctcaccc 600
gtgctgagac tgtttttcct gatgtggact atgtaaattc tactgaagct gaaaccattt 660
tggataacat cactcaaagc acccaatcat ttaatgactt cactcgggtt gttggtggag 720
aagatgccaa accaggtcaa ttcccttggc aggttgtttt gaatggtaaa gttgatgcat 780
tctgtggagg ctctatcgtt aatgaaaaat ggattgtaac tgctgcccac tgtgttgaaa 840
ctggtgttaa aattacagtt gtcgcaggtg aacataatat tgaggagaca gaacatacag 900
agcaaaagcg aaatgtgatt cgaattattc ctcaccacaa ctacaatgca gctattaata 960
agtacaacca tgacattgcc cttctggaac tggacgaacc cttagtgcta aacagctacg 1020
ttacacctat ttgcattgct gacaaggaat acacgaacat cttcctcaaa tttggatctg 1080
gctatgtaag tggctgggga agagtcttcc acaaagggag atcagcttta gttcttcagt 1140
accttagagt tccacttgtt gaccgagcca catgtcttcg atctacaaag ttcaccatct 1200
ataacaacat gttctgtgct ggcttccatg aaggaggtag agattcatgt caaggagata 1260
gtgggggacc ccatgttact gaagtggaag ggaccagttt cttaactgga attattagct 1320
ggggtgaaga gtgtgcaatg aaaggcaaat atggaatata taccaaggta tcccggtatg 1380
tcaactggat taaggaaaaa acaaagctca cttaatgaaa gatggatttc caaggttaat 1440
tcattggaat tgaaaattaa cagggcctct cactaactaa tcactttccc atcttttgtt 1500
agatttgaat atatacattc tatgatcatt gctttttctc tttacagggg agaatttcat 1560
attttacctg agcaaattga ttagaaaatg gaaccactag aggaatataa tgtgttagga 1620
aattacagtc atttctaagg gcccagccct tgacaaaatt gtgaagttaa attctccact 1680
ctgtccatca gatactatgg ttctccacta tggcaactaa ctcactcaat tttccctcct 1740
tagcagcatt ccatcttccc gatcttcttt gcttctccaa ccaaaacatc aatgtttatt 1800
agttctgtat acagtacagg atctttggtc tactctatca caaggccagt accacactca 1860
tgaagaaaga acacaggagt agctgagagg ctaaaactca tcaaaaacac tactcctttt 1920
cctctaccct attcctcaat cttttacctt ttccaaatcc caatccccaa atcagttttt 1980
ctctttctta ctccctctct cccttttacc ctccatggtc gttaaaggag agatggggag 2040
catcattctg ttatacttct gtacacagtt atacatgtct atcaaaccca gacttgcttc 2100
catagtggag acttgctttt cagaacatag ggatgaagta aggtgcctga aaagtttggg 2160
ggaaaagttt ctttcagaga gttaagttat tttatatata taatatatat ataaaatata 2220
taatatacaa tataaatata tagtgtgtgt gtgtatgcgt gtgtgtagac acacacgcat 2280
acacacatat aatggaagca ataagccatt ctaagagctt gtatggttat ggaggtctga 2340
ctaggcatga tttcacgaag gcaagattgg catatcattg taactaaaaa agctgacatt 2400
gacccagaca tattgtactc tttctaaaaa taataataat aatgctaaca gaaagaagag 2460
aaccgttcgt ttgcaatcta cagctagtag agactttgag gaagaattca acagtgtgtc 2520
ttcagcagtg ttcagagcca agcaagaagt tgaagttgcc tagaccagag gacataagta 2580
tcatgtctcc tttaactagc ataccccgaa gtggagaagg gtgcagcagg ctcaaaggca 2640
taagtcattc caatcagcca actaagttgt ccttttctgg tttcgtgttc accatggaac 2700
attttgatta tagttaatcc ttctatcttg aatcttctag agagttgctg accaactgac 2760
gtatgtttcc ctttgtgaat taataaactg gtgttctggt tcat 2804
<210>171
<211>461
<212>PRT
<213>Homo sapiens
<220>
<223>coagulation factor IXa
<400>171
Met Gln Arg Val Asn Met Ile Met Ala Glu Ser Pro Gly Leu Ile Thr
1 5 10 15
Ile Cys Leu Leu Gly Tyr Leu Leu Ser Ala Glu Cys Thr Val Phe Leu
20 25 30
Asp His Glu Asn Ala Asn Lys Ile Leu Asn Arg Pro Lys Arg Tyr Asn
35 40 45
Ser Gly Lys Leu Glu Glu Phe Val Gln Gly Asn Leu Glu Arg Glu Cys
50 55 60
Met Glu Glu Lys Cys Ser Phe Glu Glu Ala Arg Glu Val Phe Glu Asn
65 70 75 80
Thr Glu Arg Thr Thr Glu Phe Trp Lys Gln Tyr Val Asp Gly Asp Gln
85 90 95
Cys Glu Ser Asn Pro Cys Leu Asn Gly Gly Ser Cys Lys Asp Asp Ile
100 105 110
Asn Ser Tyr Glu Cys Trp Cys Pro Phe Gly Phe Glu Gly Lys Asn Cys
115 120 125
Glu Leu Asp Val Thr Cys Asn Ile Lys Asn Gly Arg Cys Glu Gln Phe
130 135 140
Cys Lys Asn Ser Ala Asp Asn Lys Val Val Cys Ser Cys Thr Glu Gly
145 150 155 160
Tyr Arg Leu Ala Glu Asn Gln Lys Ser Cys Glu Pro Ala Val Pro Phe
165 170 175
Pro Cys Gly Arg Val Ser Val Ser Gln Thr Ser Lys Leu Thr Arg Ala
180 185 190
Glu Thr Val Phe Pro Asp Val Asp Tyr Val Asn Ser Thr Glu Ala Glu
195 200 205
Thr Ile Leu Asp Asn Ile Thr Gln Ser Thr Gln Ser Phe Asn Asp Phe
210 215 220
Thr Arg Val Val Gly Gly Glu Asp Ala Lys Pro Gly Gln Phe Pro Trp
225 230 235 240
Gln Val Val Leu Asn Gly Lys Val Asp Ala Phe Cys Gly Gly Ser Ile
245 250 255
Val Asn Glu Lys Trp Ile Val Thr Ala Ala His Cys Val Glu Thr Gly
260 265 270
Val Lys Ile Thr Val Val Ala Gly Glu His Asn Ile Glu Glu Thr Glu
275 280 285
His Thr Glu Gln Lys Arg Asn Val Ile Arg Ile Ile Pro His His Asn
290 295 300
Tyr Asn Ala Ala Ile Asn Lys Tyr Asn His Asp Ile Ala Leu Leu Glu
305 310 315 320
Leu Asp Glu Pro Leu Val Leu Asn Ser Tyr Val Thr Pro Ile Cys Ile
325 330 335
Ala Asp Lys Glu Tyr Thr Asn Ile Phe Leu Lys Phe Gly Ser Gly Tyr
340 345 350
Val Ser Gly Trp Gly Arg Val Phe His Lys Gly Arg Ser Ala Leu Val
355 360 365
Leu Gln Tyr Leu Arg Val Pro Leu Val Asp Arg Ala Thr Cys Leu Arg
370 375 380
Ser Thr Lys Phe Thr Ile Tyr Asn Asn Met Phe Cys Ala Gly Phe His
385 390 395 400
Glu Gly Gly Arg Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro His Val
405 410 415
Thr Glu Val Glu Gly Thr Ser Phe Leu Thr Gly Ile Ile Ser Trp Gly
420 425 430
Glu Glu Cys Ala Met Lys Gly Lys Tyr Gly Ile Tyr Thr Lys Val Ser
435 440 445
Arg Tyr Val Asn Trp Ile Lys Glu Lys Thr Lys Leu Thr
450 455 460
<210>172
<211>2478
<212>DNA
<213>Homo sapiens
<220>
<223>coagulation factor VIIa transcript variant 1
<400>172
agtcccatgg ggaatgtcaa caggcagggg cagcactgca gagatttcat catggtctcc 60
caggccctca ggctcctctg ccttctgctt gggcttcagg gctgcctggc tgcaggcggg 120
gtcgctaagg cctcaggagg agaaacacgg gacatgccgt ggaagccggg gcctcacaga 180
gtcttcgtaa cccaggagga agcccacggc gtcctgcacc ggcgccggcg cgccaacgcg 240
ttcctggagg agctgcggcc gggctccctg gagagggagt gcaaggagga gcagtgctcc 300
ttcgaggagg cccgggagat cttcaaggac gcggagagga cgaagctgtt ctggatttct 360
tacagtgatg gggaccagtg tgcctcaagt ccatgccaga atgggggctc ctgcaaggac 420
cagctccagt cctatatctg cttctgcctc cctgccttcg agggccggaa ctgtgagacg 480
cacaaggatg accagctgat ctgtgtgaac gagaacggcg gctgtgagca gtactgcagt 540
gaccacacgg gcaccaagcg ctcctgtcgg tgccacgagg ggtactctct gctggcagac 600
ggggtgtcct gcacacccac agttgaatat ccatgtggaa aaatacctat tctagaaaaa 660
agaaatgcca gcaaacccca aggccgaatt gtggggggca aggtgtgccc caaaggggag 720
tgtccatggc aggtcctgtt gttggtgaat ggagctcagt tgtgtggggg gaccctgatc 780
aacaccatct gggtggtctc cgcggcccac tgtttcgaca aaatcaagaa ctggaggaac 840
ctgatcgcgg tgctgggcga gcacgacctc agcgagcacg acggggatga gcagagccgg 900
cgggtggcgc aggtcatcat ccccagcacg tacgtcccgg gcaccaccaa ccacgacatc 960
gcgctgctcc gcctgcacca gcccgtggtc ctcactgacc atgtggtgcc cctctgcctg 1020
cccgaacgga cgttctctga gaggacgctg gccttcgtgc gcttctcatt ggtcagcggc 1080
tggggccagc tgctggaccg tggcgccacg gccctggagc tcatggtgct caacgtgccc 1140
cggctgatga cccaggactg cctgcagcag tcacggaagg tgggagactc cccaaatatc 1200
acggagtaca tgttctgtgc cggctactcg gatggcagca aggactcctg caagggggac 1260
agtggaggcc cacatgccac ccactaccgg ggcacgtggt acctgacggg catcgtcagc 1320
tggggccagg gctgcgcaac cgtgggccac tttggggtgt acaccagggt ctcccagtac 1380
atcgagtggc tgcaaaagct catgcgctca gagccacgcc caggagtcct cctgcgagcc 1440
ccatttccct agcccagcag ccctggcctg tggagagaaa gccaaggctg cgtcgaactg 1500
tcctggcacc aaatcccata tattcttctg cagttaatgg ggtagaggag ggcatgggag 1560
ggagggagag gtggggaggg agacagagac agaaacagag agagacagag acagagagag 1620
actgagggag agactctgag gacatggaga gagactcaaa gagactccaa gattcaaaga 1680
gactaataga gacacagaga tggaatagaa aagatgagag gcagaggcag acaggcgctg 1740
gacagagggg caggggagtg ccaaggttgt cctggaggca gacagcccag ctgagcctcc 1800
ttacctccct tcagccaagc cccacctgca cgtgatctgc tggccctcag gctgctgctc 1860
tgccttcatt gctggagaca gtagaggcat gaacacacat ggatgcacac acacacacgc 1920
caatgcacac acacagagat atgcacacac acggatgcac acacagatgg tcacacagag 1980
atacgcaaac acaccgatgc acacgcacat agagatatgc acacacagat gcacacacag 2040
atatacacat ggatgcacgc acatgccaat gcacgcacac atcagtgcac acggatgcac 2100
agagatatgc acacaccgat gtgcgcacac acagatatgc acacacatgg atgagcacac 2160
acacaccaag tgcgcacaca caccgatgta cacacacaga tgcacacaca gatgcacaca 2220
caccgatgct gactccatgt gtgctgtcct ctgaaggcgg ttgtttagct ctcacttttc 2280
tggttcttat ccattatcat cttcacttca gacaattcag aagcatcacc atgcatggtg 2340
gcgaatgccc ccaaactctc ccccaaatgt atttctccct tcgctgggtg ccgggctgca 2400
cagactattc cccacctgct tcccagcttc acaataaacg gctgcgtctc ctccgcacac 2460
ctgtggtgcc tgccaccc 2478
<210>173
<211>466
<212>PRT
<213>Homo sapiens
<220>
<223>coagulation factor VIIa isoform 1
<400>173
Met Val Ser Gln Ala Leu Arg Leu Leu Cys Leu Leu Leu Gly Leu Gln
1 5 10 15
Gly Cys Leu Ala Ala Gly Gly Val Ala Lys Ala Ser Gly Gly Glu Thr
20 25 30
Arg Asp Met Pro Trp Lys Pro Gly Pro His Arg Val Phe Val Thr Gln
35 40 45
Glu Glu Ala His Gly Val Leu His Arg Arg Arg Arg Ala Asn Ala Phe
50 55 60
Leu Glu Glu Leu Arg Pro Gly Ser Leu Glu Arg Glu Cys Lys Glu Glu
65 70 75 80
Gln Cys Ser Phe Glu Glu Ala Arg Glu Ile Phe Lys Asp Ala Glu Arg
85 90 95
Thr Lys Leu Phe Trp Ile Ser Tyr Ser Asp Gly Asp Gln Cys Ala Ser
100 105 110
Ser Pro Cys Gln Asn Gly Gly Ser Cys Lys Asp Gln Leu Gln Ser Tyr
115 120 125
Ile Cys Phe Cys Leu Pro Ala Phe Glu Gly Arg Asn Cys Glu Thr His
130 135 140
Lys Asp Asp Gln Leu Ile Cys Val Asn Glu Asn Gly Gly Cys Glu Gln
145 150 155 160
Tyr Cys Ser Asp His Thr Gly Thr Lys Arg Ser Cys Arg Cys His Glu
165 170 175
Gly Tyr Ser Leu Leu Ala Asp Gly Val Ser Cys Thr Pro Thr Val Glu
180 185 190
Tyr Pro Cys Gly Lys Ile Pro Ile Leu Glu Lys Arg Asn Ala Ser Lys
195 200 205
Pro Gln Gly Arg Ile Val Gly Gly Lys Val Cys Pro Lys Gly Glu Cys
210 215 220
Pro Trp Gln Val Leu Leu Leu Val Asn Gly Ala Gln Leu Cys Gly Gly
225 230 235 240
Thr Leu Ile Asn Thr Ile Trp Val Val Ser Ala Ala His Cys Phe Asp
245 250 255
Lys Ile Lys Asn Trp Arg Asn Leu Ile Ala Val Leu Gly Glu His Asp
260 265 270
Leu Ser Glu His Asp Gly Asp Glu Gln Ser Arg Arg Val Ala Gln Val
275 280 285
Ile Ile Pro Ser Thr Tyr Val Pro Gly Thr Thr Asn His Asp Ile Ala
290 295 300
Leu Leu Arg Leu His Gln Pro Val Val Leu Thr Asp His Val Val Pro
305 310 315 320
Leu Cys Leu Pro Glu Arg Thr Phe Ser Glu Arg Thr Leu Ala Phe Val
325 330 335
Arg Phe Ser Leu Val Ser Gly Trp Gly Gln Leu Leu Asp Arg Gly Ala
340 345 350
Thr Ala Leu Glu Leu Met Val Leu Asn Val Pro Arg Leu Met Thr Gln
355 360 365
Asp Cys Leu Gln Gln Ser Arg Lys Val Gly Asp Ser Pro Asn Ile Thr
370 375 380
Glu Tyr Met Phe Cys Ala Gly Tyr Ser Asp Gly Ser Lys Asp Ser Cys
385 390 395 400
Lys Gly Asp Ser Gly Gly Pro His Ala Thr His Tyr Arg Gly Thr Trp
405 410 415
Tyr Leu Thr Gly Ile Val Ser Trp Gly Gln Gly Cys Ala Thr Val Gly
420 425 430
His Phe Gly Val Tyr Thr Arg Val Ser Gln Tyr Ile Glu Trp Leu Gln
435 440 445
Lys Leu Met Arg Ser Glu Pro Arg Pro Gly Val Leu Leu Arg Ala Pro
450 455 460
Phe Pro
465
<210>174
<211>2412
<212>DNA
<213>Homo sapiens
<220>
<223>coagulation factor VIIa transcript variant 2
<400>174
agtcccatgg ggaatgtcaa caggcagggg cagcactgca gagatttcat catggtctcc 60
caggccctca ggctcctctg ccttctgctt gggcttcagg gctgcctggc tgcagtcttc 120
gtaacccagg aggaagccca cggcgtcctg caccggcgcc ggcgcgccaa cgcgttcctg 180
gaggagctgc ggccgggctc cctggagagg gagtgcaagg aggagcagtg ctccttcgag 240
gaggcccggg agatcttcaa ggacgcggag aggacgaagc tgttctggat ttcttacagt 300
gatggggacc agtgtgcctc aagtccatgc cagaatgggg gctcctgcaa ggaccagctc 360
cagtcctata tctgcttctg cctccctgcc ttcgagggcc ggaactgtga gacgcacaag 420
gatgaccagc tgatctgtgt gaacgagaac ggcggctgtg agcagtactg cagtgaccac 480
acgggcacca agcgctcctg tcggtgccac gaggggtact ctctgctggc agacggggtg 540
tcctgcacac ccacagttga atatccatgt ggaaaaatac ctattctaga aaaaagaaat 600
gccagcaaac cccaaggccg aattgtgggg ggcaaggtgt gccccaaagg ggagtgtcca 660
tggcaggtcc tgttgttggt gaatggagct cagttgtgtg gggggaccct gatcaacacc 720
atctgggtgg tctccgcggc ccactgtttc gacaaaatca agaactggag gaacctgatc 780
gcggtgctgg gcgagcacga cctcagcgag cacgacgggg atgagcagag ccggcgggtg 840
gcgcaggtca tcatccccag cacgtacgtc ccgggcacca ccaaccacga catcgcgctg 900
ctccgcctgc accagcccgt ggtcctcact gaccatgtgg tgcccctctg cctgcccgaa 960
cggacgttct ctgagaggac gctggccttc gtgcgcttct cattggtcag cggctggggc 1020
cagctgctgg accgtggcgc cacggccctg gagctcatgg tgctcaacgt gccccggctg 1080
atgacccagg actgcctgca gcagtcacgg aaggtgggag actccccaaa tatcacggag 1140
tacatgttct gtgccggcta ctcggatggc agcaaggact cctgcaaggg ggacagtgga 1200
ggcccacatg ccacccacta ccggggcacg tggtacctga cgggcatcgt cagctggggc 1260
cagggctgcg caaccgtggg ccactttggg gtgtacacca gggtctccca gtacatcgag 1320
tggctgcaaa agctcatgcg ctcagagcca cgcccaggag tcctcctgcg agccccattt 1380
ccctagccca gcagccctgg cctgtggaga gaaagccaag gctgcgtcga actgtcctgg 1440
caccaaatcc catatattct tctgcagtta atggggtaga ggagggcatg ggagggaggg 1500
agaggtgggg agggagacag agacagaaac agagagagac agagacagag agagactgag 1560
ggagagactc tgaggacatg gagagagact caaagagact ccaagattca aagagactaa 1620
tagagacaca gagatggaat agaaaagatg agaggcagag gcagacaggc gctggacaga 1680
ggggcagggg agtgccaagg ttgtcctgga ggcagacagc ccagctgagc ctccttacct 1740
cccttcagcc aagccccacc tgcacgtgat ctgctggccc tcaggctgct gctctgcctt 1800
cattgctgga gacagtagag gcatgaacac acatggatgc acacacacac acgccaatgc 1860
acacacacag agatatgcac acacacggat gcacacacag atggtcacac agagatacgc 1920
aaacacaccg atgcacacgc acatagagat atgcacacac agatgcacac acagatatac 1980
acatggatgc acgcacatgc caatgcacgc acacatcagt gcacacggat gcacagagat 2040
atgcacacac cgatgtgcgc acacacagat atgcacacac atggatgagc acacacacac 2100
caagtgcgca cacacaccga tgtacacaca cagatgcaca cacagatgca cacacaccga 2160
tgctgactcc atgtgtgctg tcctctgaag gcggttgttt agctctcact tttctggttc 2220
ttatccatta tcatcttcac ttcagacaat tcagaagcat caccatgcat ggtggcgaat 2280
gcccccaaac tctcccccaa atgtatttct cccttcgctg ggtgccgggc tgcacagact 2340
attccccacc tgcttcccag cttcacaata aacggctgcg tctcctccgc acacctgtgg 2400
tgcctgccac cc 2412
<210>175
<211>444
<212>PRT
<213>Homo sapiens
<220>
<223>coagulation factor VIIa isoform 2
<400>175
Met Val Ser Gln Ala Leu Arg Leu Leu Cys Leu Leu Leu Gly Leu Gln
1 5 10 15
Gly Cys Leu Ala Ala Val Phe Val Thr Gln Glu Glu Ala His Gly Val
20 25 30
Leu His Arg Arg Arg Arg Ala Asn Ala Phe Leu Glu Glu Leu Arg Pro
35 40 45
Gly Ser Leu Glu Arg Glu Cys Lys Glu Glu Gln Cys Ser Phe Glu Glu
50 55 60
Ala Arg Glu Ile Phe Lys Asp Ala Glu Arg Thr Lys Leu Phe Trp Ile
65 70 75 80
Ser Tyr Ser Asp Gly Asp Gln Cys Ala Ser Ser Pro Cys Gln Asn Gly
85 90 95
Gly Ser Cys Lys Asp Gln Leu Gln Ser Tyr Ile Cys Phe Cys Leu Pro
100 105 110
Ala Phe Glu Gly Arg Asn Cys Glu Thr His Lys Asp Asp Gln Leu Ile
115 120 125
Cys Val Asn Glu Asn Gly Gly Cys Glu Gln Tyr Cys Ser Asp His Thr
130 135 140
Gly Thr Lys Arg Ser Cys Arg Cys His Glu Gly Tyr Ser Leu Leu Ala
145 150 155 160
Asp Gly Val Ser Cys Thr Pro Thr Val Glu Tyr Pro Cys Gly Lys Ile
165 170 175
Pro Ile Leu Glu Lys Arg Asn Ala Ser Lys Pro Gln Gly Arg Ile Val
180 185 190
Gly Gly Lys Val Cys Pro Lys Gly Glu Cys Pro Trp Gln Val Leu Leu
195 200 205
Leu Val Asn Gly Ala Gln Leu Cys Gly Gly Thr Leu Ile Asn Thr Ile
210 215 220
Trp Val Val Ser Ala Ala His Cys Phe Asp Lys Ile Lys Asn Trp Arg
225 230 235 240
Asn Leu Ile Ala Val Leu Gly Glu His Asp Leu Ser Glu His Asp Gly
245 250 255
Asp Glu Gln Ser Arg Arg Val Ala Gln Val Ile Ile Pro Ser Thr Tyr
260 265 270
Val Pro Gly Thr Thr Asn His Asp Ile Ala Leu Leu Arg Leu His Gln
275 280 285
Pro Val Val Leu Thr Asp His Val Val Pro Leu Cys Leu Pro Glu Arg
290 295 300
Thr Phe Ser Glu Arg Thr Leu Ala Phe Val Arg Phe Ser Leu Val Ser
305 310 315 320
Gly Trp Gly Gln Leu Leu Asp Arg Gly Ala Thr Ala Leu Glu Leu Met
325 330 335
Val Leu Asn Val Pro Arg Leu Met Thr Gln Asp Cys Leu Gln Gln Ser
340 345 350
Arg Lys Val Gly Asp Ser Pro Asn Ile Thr Glu Tyr Met Phe Cys Ala
355 360 365
Gly Tyr Ser Asp Gly Ser Lys Asp Ser Cys Lys Gly Asp Ser Gly Gly
370 375 380
Pro His Ala Thr His Tyr Arg Gly Thr Trp Tyr Leu Thr Gly Ile Val
385 390 395 400
Ser Trp Gly Gln Gly Cys Ala Thr Val Gly His Phe Gly Val Tyr Thr
405 410 415
Arg Val Ser Gln Tyr Ile Glu Trp Leu Gln Lys Leu Met Arg Ser Glu
420 425 430
Pro Arg Pro Gly Val Leu Leu Arg Ala Pro Phe Pro
435 440
<210>176
<211>1502
<212>DNA
<213>Homo sapiens
<220>
<223>coagulation factor Xa
<400>176
cagggacaca gtactcggcc acaccatggg gcgcccactg cacctcgtcc tgctcagtgc 60
ctccctggct ggcctcctgc tgctcgggga aagtctgttc atccgcaggg agcaggccaa 120
caacatcctg gcgagggtca cgagggccaa ttcctttctt gaagagatga agaaaggaca 180
cctcgaaaga gagtgcatgg aagagacctg ctcatacgaa gaggcccgcg aggtctttga 240
ggacagcgac aagacgaatg aattctggaa taaatacaaa gatggcgacc agtgtgagac 300
cagtccttgc cagaaccagg gcaaatgtaa agacggcctc ggggaataca cctgcacctg 360
tttagaagga ttcgaaggca aaaactgtga attattcaca cggaagctct gcagcctgga 420
caacggggac tgtgaccagt tctgccacga ggaacagaac tctgtggtgt gctcctgcgc 480
ccgcgggtac accctggctg acaacggcaa ggcctgcatt cccacagggc cctacccctg 540
tgggaaacag accctggaac gcaggaagag gtcagtggcc caggccacca gcagcagcgg 600
ggaggcccct gacagcatca catggaagcc atatgatgca gccgacctgg accccaccga 660
gaaccccttc gacctgcttg acttcaacca gacgcagcct gagaggggcg acaacaacct 720
caccaggatc gtgggaggcc aggaatgcaa ggacggggag tgtccctggc aggccctgct 780
catcaatgag gaaaacgagg gtttctgtgg tggaaccatt ctgagcgagt tctacatcct 840
aacggcagcc cactgtctct accaagccaa gagattcaag gtgagggtag gggaccggaa 900
cacggagcag gaggagggcg gtgaggcggt gcacgaggtg gaggtggtca tcaagcacaa 960
ccggttcaca aaggagacct atgacttcga catcgccgtg ctccggctca agacccccat 1020
caccttccgc atgaacgtgg cgcctgcctg cctccccgag cgtgactggg ccgagtccac 1080
gctgatgacg cagaagacgg ggattgtgag cggcttcggg cgcacccacg agaagggccg 1140
gcagtccacc aggctcaaga tgctggaggt gccctacgtg gaccgcaaca gctgcaagct 1200
gtccagcagc ttcatcatca cccagaacat gttctgtgcc ggctacgaca ccaagcagga 1260
ggatgcctgc cagggggaca gcgggggccc gcacgtcacc cgcttcaagg acacctactt 1320
cgtgacaggc atcgtcagct ggggagaggg ctgtgcccgt aaggggaagt acgggatcta 1380
caccaaggtc accgccttcc tcaagtggat cgacaggtcc atgaaaacca ggggcttgcc 1440
caaggccaag agccatgccc cggaggtcat aacgtcctct ccattaaagt gagatcccac 1500
tc 1502
<210>177
<211>488
<212>PRT
<213>Homo sapiens
<220>
<223>coagulation factor Xa
<400>177
Met Gly Arg Pro Leu His Leu Val Leu Leu Ser Ala Ser Leu Ala Gly
1 5 10 15
Leu Leu Leu Leu Gly Glu Ser Leu Phe Ile Arg Arg Glu Gln Ala Asn
20 25 30
Asn Ile Leu Ala Arg Val Thr Arg Ala Asn Ser Phe Leu Glu Glu Met
35 40 45
Lys Lys Gly His Leu Glu Arg Glu Cys Met Glu Glu Thr Cys Ser Tyr
50 55 60
Glu Glu Ala Arg Glu Val Phe Glu Asp Ser Asp Lys Thr Asn Glu Phe
65 70 75 80
Trp Asn Lys Tyr Lys Asp Gly Asp Gln Cys Glu Thr Ser Pro Cys Gln
85 90 95
Asn Gln Gly Lys Cys Lys Asp Gly Leu Gly Glu Tyr Thr Cys Thr Cys
100 105 110
Leu Glu Gly Phe Glu Gly Lys Asn Cys Glu Leu Phe Thr Arg Lys Leu
115 120 125
Cys Ser Leu Asp Asn Gly Asp Cys Asp Gln Phe Cys His Glu Glu Gln
130 135 140
Asn Ser Val Val Cys Ser Cys Ala Arg Gly Tyr Thr Leu Ala Asp Asn
145 150 155 160
Gly Lys Ala Cys Ile Pro Thr Gly Pro Tyr Pro Cys Gly Lys Gln Thr
165 170 175
Leu Glu Arg Arg Lys Arg Ser Val Ala Gln Ala Thr Ser Ser Ser Gly
180 185 190
Glu Ala Pro Asp Ser Ile Thr Trp Lys Pro Tyr Asp Ala Ala Asp Leu
195 200 205
Asp Pro Thr Glu Asn Pro Phe Asp Leu Leu Asp Phe Asn Gln Thr Gln
210 215 220
Pro Glu Arg Gly Asp Asn Asn Leu Thr Arg Ile Val Gly Gly Gln Glu
225 230 235 240
Cys Lys Asp Gly Glu Cys Pro Trp Gln Ala Leu Leu Ile Asn Glu Glu
245 250 255
Asn Glu Gly Phe Cys Gly Gly Thr Ile Leu Ser Glu Phe Tyr Ile Leu
260 265 270
Thr Ala Ala His Cys Leu Tyr Gln Ala Lys Arg Phe Lys Val Arg Val
275 280 285
Gly Asp Arg Asn Thr Glu Gln Glu Glu Gly Gly Glu Ala Val His Glu
290 295 300
Val Glu Val Val Ile Lys His Asn Arg Phe Thr Lys Glu Thr Tyr Asp
305 310 315 320
Phe Asp Ile Ala Val Leu Arg Leu Lys Thr Pro Ile Thr Phe Arg Met
325 330 335
Asn Val Ala Pro Ala Cys Leu Pro Glu Arg Asp Trp Ala Glu Ser Thr
340 345 350
Leu Met Thr Gln Lys Thr Gly Ile Val Ser Gly Phe Gly Arg Thr His
355 360 365
Glu Lys Gly Arg Gln Ser Thr Arg Leu Lys Met Leu Glu Val Pro Tyr
370 375 380
Val Asp Arg Asn Ser Cys Lys Leu Ser Ser Ser Phe Ile Ile Thr Gln
385 390 395 400
Asn Met Phe Cys Ala Gly Tyr Asp Thr Lys Gln Glu Asp Ala Cys Gln
405 410 415
Gly Asp Ser Gly Gly Pro His Val Thr Arg Phe Lys Asp Thr Tyr Phe
420 425 430
Val Thr Gly Ile Val Ser Trp Gly Glu Gly Cys Ala Arg Lys Gly Lys
435 440 445
Tyr Gly Ile Tyr Thr Lys Val Thr Ala Phe Leu Lys Trp Ile Asp Arg
450 455 460
Ser Met Lys Thr Arg Gly Leu Pro Lys Ala Lys Ser His Ala Pro Glu
465 470 475 480
Val Ile Thr Ser Ser Pro Leu Lys
485
<210>178
<211>1997
<212>DNA
<213>Homo sapiens
<220>
<223>Thrombin
<400>178
aattcctcag tgacccagga gctgacacac tatggcgcac gtccgaggct tgcagctgcc 60
tggctgcctg gccctggctg ccctgtgtag ccttgtgcac agccagcatg tgttcctggc 120
tcctcagcaa gcacggtcgc tgctccagcg ggtccggcga gccaacacct tcttggagga 180
ggtgcgcaag ggcaacctag agcgagagtg cgtggaggag acgtgcagct acgaggaggc 240
cttcgaggct ctggagtcct ccacggctac ggatgtgttc tgggccaagt acacagcttg 300
tgagacagcg aggacgcctc gagataagct tgctgcatgt ctggaaggta actgtgctga 360
gggtctgggt acgaactacc gagggcatgt gaacatcacc cggtcaggca ttgagtgcca 420
gctatggagg agtcgctacc cacataagcc tgaaatcaac tccactaccc atcctggggc 480
cgacctacag gagaatttct gccgcaaccc cgacagcagc accacgggac cctggtgcta 540
cactacagac cccaccgtga ggaggcagga atgcagcatc cctgtctgtg gccaggatca 600
agtcactgta gcgatgactc cacgctccga aggctccagt gtgaatctgt cacctccatt 660
ggagcagtgt gtccctgatc gggggcagca gtaccagggg cgcctggcgg tgaccacaca 720
tgggctcccc tgcctggcct gggccagcgc acaggccaag gccctgagca agcaccagga 780
cttcaactca gctgtgcagc tggtggagaa cttctgccgc aacccagacg gggatgagga 840
gggcgtgtgg tgctatgtgg ccgggaagcc tggcgacttt gggtactgcg acctcaacta 900
ttgtgaggag gccgtggagg aggagacagg agatgggctg gatgaggact cagacagggc 960
catcgaaggg cgtaccgcca ccagtgagta ccagactttc ttcaatccga ggacctttgg 1020
ctcgggagag gcagactgtg ggctgcgacc tctgttcgag aagaagtcgc tggaggacaa 1080
aaccgaaaga gagctcctgg aatcctacat cgacgggcgc attgtggagg gctcggatgc 1140
agagatcggc atgtcacctt ggcaggtgat gcttttccgg aagagtcccc aggagctgct 1200
gtgtggggcc agcctcatca gtgaccgctg ggtcctcacc gccgcccact gcctcctgta 1260
cccgccctgg gacaagaact tcaccgagaa tgaccttctg gtgcgcattg gcaagcactc 1320
ccgcacaagg tacgagcgaa acattgaaaa gatatccatg ttggaaaaga tctacatcca 1380
ccccaggtac aactggcggg agaacctgga ccgggacatt gccctgatga agctgaagaa 1440
gcctgttgcc ttcagtgact acattcaccc tgtgtgtctg cccgacaggg agacggcagc 1500
cagcttgctc caggctggat acaaggggcg ggtgacaggc tggggcaacc tgaaggagac 1560
gtggacagcc aacgttggta aggggcagcc cagtgtcctg caggtggtga acctgcccat 1620
tgtggagcgg ccggtctgca aggactccac ccggatccgc atcactgaca acatgttctg 1680
tgctggttac aagcctgatg aagggaaacg aggggatgcc tgtgaaggtg acagtggggg 1740
accctttgtc atgaagagcc cctttaacaa ccgctggtat caaatgggca tcgtctcatg 1800
gggtgaaggc tgtgaccggg atgggaaata tggcttctac acacatgtgt tccgcctgaa 1860
gaagtggata cagaaggtca ttgatcagtt tggagagtag gggccactca tattctgggc 1920
tcctggaacc aatcccgtga aagaattatt tttgtgtttc taaaactatg gttcccaata 1980
aaagtgactc tcagcgg 1997
<210>179
<211>622
<212>PRT
<213>Homo sapiens
<220>
<223>Thrombin
<400>179
Met Ala His Val Arg Gly Leu Gln Leu Pro Gly Cys Leu Ala Leu Ala
1 5 10 15
Ala Leu Cys Ser Leu Val His Ser Gln His Val Phe Leu Ala Pro Gln
20 25 30
Gln Ala Arg Ser Leu Leu Gln Arg Val Arg Arg Ala Asn Thr Phe Leu
35 40 45
Glu Glu Val Arg Lys Gly Asn Leu Glu Arg Glu Cys Val Glu Glu Thr
50 55 60
Cys Ser Tyr Glu Glu Ala Phe Glu Ala Leu Glu Ser Ser Thr Ala Thr
65 70 75 80
Asp Val Phe Trp Ala Lys Tyr Thr Ala Cys Glu Thr Ala Arg Thr Pro
85 90 95
Arg Asp Lys Leu Ala Ala Cys Leu Glu Gly Asn Cys Ala Glu Gly Leu
100 105 110
Gly Thr Asn Tyr Arg Gly His Val Asn Ile Thr Arg Ser Gly Ile Glu
115 120 125
Cys Gln Leu Trp Arg Ser Arg Tyr Pro His Lys Pro Glu Ile Asn Ser
130 135 140
Thr Thr His Pro Gly Ala Asp Leu Gln Glu Asn Phe Cys Arg Asn Pro
145 150 155 160
Asp Ser Ser Thr Thr Gly Pro Trp Cys Tyr Thr Thr Asp Pro Thr Val
165 170 175
Arg Arg Gln Glu Cys Ser Ile Pro Val Cys Gly Gln Asp Gln Val Thr
180 185 190
Val Ala Met Thr Pro Arg Ser Glu Gly Ser Ser Val Asn Leu Ser Pro
195 200 205
Pro Leu Glu Gln Cys Val Pro Asp Arg Gly Gln Gln Tyr Gln Gly Arg
210 215 220
Leu Ala Val Thr Thr His Gly Leu Pro Cys Leu Ala Trp Ala Ser Ala
225 230 235 240
Gln Ala Lys Ala Leu Ser Lys His Gln Asp Phe Asn Ser Ala Val Gln
245 250 255
Leu Val Glu Asn Phe Cys Arg Asn Pro Asp Gly Asp Glu Glu Gly Val
260 265 270
Trp Cys Tyr Val Ala Gly Lys Pro Gly Asp Phe Gly Tyr Cys Asp Leu
275 280 285
Asn Tyr Cys Glu Glu Ala Val Glu Glu Glu Thr Gly Asp Gly Leu Asp
290 295 300
Glu Asp Ser Asp Arg Ala Ile Glu Gly Arg Thr Ala Thr Ser Glu Tyr
305 310 315 320
Gln Thr Phe Phe Asn Pro Arg Thr Phe Gly Ser Gly Glu Ala Asp Cys
325 330 335
Gly Leu Arg Pro Leu Phe Glu Lys Lys Ser Leu Glu Asp Lys Thr Glu
340 345 350
Arg Glu Leu Leu Glu Ser Tyr Ile Asp Gly Arg Ile Val Glu Gly Ser
355 360 365
Asp Ala Glu Ile Gly Met Ser Pro Trp Gln Val Met Leu Phe Arg Lys
370 375 380
Ser Pro Gln Glu Leu Leu Cys Gly Ala Ser Leu Ile Ser Asp Arg Trp
385 390 395 400
Val Leu Thr Ala Ala His Cys Leu Leu Tyr Pro Pro Trp Asp Lys Asn
405 410 415
Phe Thr Glu Asn Asp Leu Leu Val Arg Ile Gly Lys His Ser Arg Thr
420 425 430
Arg Tyr Glu Arg Asn Ile Glu Lys Ile Ser Met Leu Glu Lys Ile Tyr
435 440 445
Ile His Pro Arg Tyr Asn Trp Arg Glu Asn Leu Asp Arg Asp Ile Ala
450 455 460
Leu Met Lys Leu Lys Lys Pro Val Ala Phe Ser Asp Tyr Ile His Pro
465 470 475 480
Val Cys Leu Pro Asp Arg Glu Thr Ala Ala Ser Leu Leu Gln Ala Gly
485 490 495
Tyr Lys Gly Arg Val Thr Gly Trp Gly Asn Leu Lys Glu Thr Trp Thr
500 505 510
Ala Asn Val Gly Lys Gly Gln Pro Ser Val Leu Gln Val Val Asn Leu
515 520 525
Pro Ile Val Glu Arg Pro Val Cys Lys Asp Ser Thr Arg Ile Arg Ile
530 535 540
Thr Asp Asn Met Phe Cys Ala Gly Tyr Lys Pro Asp Glu Gly Lys Arg
545 550 555 560
Gly Asp Ala Cys Glu Gly Asp Ser Gly Gly Pro Phe Val Met Lys Ser
565 570 575
Pro Phe Asn Asn Arg Trp Tyr Gln Met Gly Ile Val Ser Trp Gly Glu
580 585 590
Gly Cys Asp Arg Asp Gly Lys Tyr Gly Phe Tyr Thr His Val Phe Arg
595 600 605
Leu Lys Lys Trp Ile Gln Lys Val Ile Asp Gln Phe Gly Glu
610 615 620
<210>180
<211>1843
<212>DNA
<213>Homo sapiens
<220>
<223>protein C (activated)
<400>180
ctgcaggggg gggggggggg gggggctgtc atggcggcag gacggcgaac ttgcagtatc 60
tccacgaccc gcccctacag gtgccagtgc ctccagaatg tggcagctca caagcctcct 120
gctgttcgtg gccacctggg gaatttccgg cacaccagct cctcttgact cagtgttctc 180
cagcagcgag cgtgcccacc aggtgctgcg gatccgcaaa cgtgccaact ccttcctgga 240
ggagctccgt cacagcagcc tggagcggga gtgcatagag gagatctgtg acttcgagga 300
ggccaaggaa attttccaaa atgtggatga cacactggcc ttctggtcca agcacgtcga 360
cggtgaccag tgcttggtct tgcccttgga gcacccgtgc gccagcctgt gctgcgggca 420
cggcacgtgc atcgacggca tcggcagctt cagctgcgac tgccgcagcg gctgggaggg 480
ccgcttctgc cagcgcgagg tgagcttcct caattgctcg ctggacaacg gcggctgcac 540
gcattactgc ctagaggagg tgggctggcg gcgctgtagc tgtgcgcctg gctacaagct 600
gggggacgac ctcctgcagt gtcaccccgc agtgaagttc ccttgtggga ggccctggaa 660
gcggatggag aagaagcgca gtcacctgaa acgagacaca gaagaccaag aagaccaagt 720
agatccgcgg ctcattgatg ggaagatgac caggcgggga gacagcccct ggcaggtggt 780
cctgctggac tcaaagaaga agctggcctg cggggcagtg ctcatccacc cctcctgggt 840
gctgacagcg gcccactgca tggatgagtc caagaagctc cttgtcaggc ttggagagta 900
tgacctgcgg cgctgggaga agtgggagct ggacctggac atcaaggagg tcttcgtcca 960
ccccaactac agcaagagca ccaccgacaa tgacatcgca ctgctgcacc tggcccagcc 1020
cgccaccctc tcgcagacca tagtgcccat ctgcctcccg gacagcggcc ttgcagagcg 1080
cgagctcaat caggccggcc aggagaccct cgtgacgggc tggggctacc acagcagccg 1140
agagaaggag gccaagagaa accgcacctt cgtcctcaac ttcatcaaga ttcccgtggt 1200
cccgcacaat gagtgcagcg aggtcatgag caacatggtg tctgagaaca tgctgtgtgc 1260
gggcatcctc ggggaccggc aggatgcctg cgagggcgac agtggggggc ccatggtcgc 1320
ctccttccac ggcacctggt tcctggtggg cctggtgagc tggggtgagg gctgtgggct 1380
ccttcacaac tacggcgttt acaccaaagt cagccgctac ctcgactgga tccatgggca 1440
catcagagac aaggaagccc cccagaagag ctgggcacct tagcgaccct ccctgcaggg 1500
ctgggctttt gcatggcaat ggatgggaca ttaaagggac atgtaacaag cacaccggcc 1560
tgctgttctg tccttccatc cctcttttgg gctcttctgg agggaagtaa catttactga 1620
gcacctgttg tatgtcacat gccttatgaa tagaatctta actcctagag caactctgtg 1680
gggtggggag gagcagatcc aagttttgcg gggtctaaag ctgtgtgtgt tgagggggat 1740
actctgttta tgaaaaagaa taaaaaacac aaccacgaaa aaaaaaaaaa aaaaaaaaaa 1800
aaaaaaaaaa aaaaaaaccc ccccccgccc cccccccctg cag 1843
<210>181
<211>461
<212>PRT
<213>Homo sapiens
<220>
<223>protein C (activated)
<400>181
Met Trp Gln Leu Thr Ser Leu Leu Leu Phe Val Ala Thr Trp Gly Ile
1 5 10 15
Ser Gly Thr Pro Ala Pro Leu Asp Ser Val Phe Ser Ser Ser Glu Arg
20 25 30
Ala His Gln Val Leu Arg Ile Arg Lys Arg Ala Asn Ser Phe Leu Glu
35 40 45
Glu Leu Arg His Ser Ser Leu Glu Arg Glu Cys Ile Glu Glu Ile Cys
50 55 60
Asp Phe Glu Glu Ala Lys Glu Ile Phe Gln Asn Val Asp Asp Thr Leu
65 70 75 80
Ala Phe Trp Ser Lys His Val Asp Gly Asp Gln Cys Leu Val Leu Pro
85 90 95
Leu Glu His Pro Cys Ala Ser Leu Cys Cys Gly His Gly Thr Cys Ile
100 105 110
Asp Gly Ile Gly Ser Phe Ser Cys Asp Cys Arg Ser Gly Trp Glu Gly
115 120 125
Arg Phe Cys Gln Arg Glu Val Ser Phe Leu Asn Cys Ser Leu Asp Asn
130 135 140
Gly Gly Cys Thr His Tyr Cys Leu Glu Glu Val Gly Trp Arg Arg Cys
145 150 155 160
Ser Cys Ala Pro Gly Tyr Lys Leu Gly Asp Asp Leu Leu Gln Cys His
165 170 175
Pro Ala Val Lys Phe Pro Cys Gly Arg Pro Trp Lys Arg Met Glu Lys
180 185 190
Lys Arg Ser His Leu Lys Arg Asp Thr Glu Asp Gln Glu Asp Gln Val
195 200 205
Asp Pro Arg Leu Ile Asp Gly Lys Met Thr Arg Arg Gly Asp Ser Pro
210 215 220
Trp Gln Val Val Leu Leu Asp Ser Lys Lys Lys Leu Ala Cys Gly Ala
225 230 235 240
Val Leu Ile His Pro Ser Trp Val Leu Thr Ala Ala His Cys Met Asp
245 250 255
Glu Ser Lys Lys Leu Leu Val Arg Leu Gly Glu Tyr Asp Leu Arg Arg
260 265 270
Trp Glu Lys Trp Glu Leu Asp Leu Asp Ile Lys Glu Val Phe Val His
275 280 285
Pro Asn Tyr Ser Lys Ser Thr Thr Asp Asn Asp Ile Ala Leu Leu His
290 295 300
Leu Ala Gln Pro Ala Thr Leu Ser Gln Thr Ile Val Pro Ile Cys Leu
305 310 315 320
Pro Asp Ser Gly Leu Ala Glu Arg Glu Leu Asn Gln Ala Gly Gln Glu
325 330 335
Thr Leu Val Thr Gly Trp Gly Tyr His Ser Ser Arg Glu Lys Glu Ala
340 345 350
Lys Arg Asn Arg Thr Phe Val Leu Asn Phe Ile Lys Ile Pro Val Val
355 360 365
Pro His Asn Glu Cys Ser Glu Val Met Ser Asn Met Val Ser Glu Asn
370 375 380
Met Leu Cys Ala Gly Ile Leu Gly Asp Arg Gln Asp Ala Cys Glu Gly
385 390 395 400
Asp Ser Gly Gly Pro Met Val Ala Ser Phe His Gly Thr Trp Phe Leu
405 410 415
Val Gly Leu Val Ser Trp Gly Glu Gly Cys Gly Leu Leu His Asn Tyr
420 425 430
Gly Val Tyr Thr Lys Val Ser Arg Tyr Leu Asp Trp Ile His Gly His
435 440 445
Ile Arg Asp Lys Glu Ala Pro Gln Lys Ser Trp Ala Pro
450 455 460
<210>182
<211>1388
<212>DNA
<213>Homo sapiens
<220>
<223>Acrosin
<400>182
caggcagtgc aggagtatgg ttgagatgct accaactgcc attctgctgg tcttggcagt 60
gtccgtggtt gctaaagata acgccacgtg tgatggcccc tgtgggttac ggttcaggca 120
aaacccacag ggtggtgtcc gcatcgtcgg cgggaaggct gcacagcatg gggcctggcc 180
ctggatggtc agcctccaga tcttcacgta caacagccac aggtaccaca catgtggagg 240
cagcttgctg aattcacgat gggtgctcac tgctgctcac tgcttcgtcg gcaaaaataa 300
tgtgcatgac tggagactgg ttttcggagc aaaggaaatt acatatggga acaataaacc 360
agtaaaggcg cctgtgcaag agagatatgt ggagaaaatc atcattcatg aaaaatacaa 420
ctctgcgaca gagggaaatg acattgccct cgtggagatc acccctccca tttcgtgtgg 480
gcgcttcatt gggccgggct gcctgcccca cttgaaggca ggcctcccca gaggctccca 540
gagctgctgg gtggccggct ggggatatat agaagagaaa gcccccaggc catcatctat 600
actgatggag gcacgtgtgg atctcatcga cctggacttg tgtaactcga cccagtggta 660
caatgggcgc gttcagccaa ccaatgtgtg cgcggggtat cctgtaggca agatcgacac 720
ctgccaggga gacagcggcg ggcctctcat gtgcaaagac agcaaggaaa gcgcctatgt 780
ggtcgtggga atcacaagct ggggggtagg ctgtgcccgt gccaagcgcc ccggaatcta 840
cacggccacc tggccttatc tgaactggat cgcctccaag attggttcta acgctttgcg 900
tatgattcaa tcggccaccc ctccaccgcc caccactcga ccgcccccga ttcgaccccc 960
cttctcccac cctatctctg ctcaccttcc ttggtatttc caaccgcccc ctcgaccact 1020
tccaccccga ccaccggcag cccagccccg acccccacct tcacccccgc ccccaccccc 1080
acctccagcc tcacctttac ccccaccccc acccccaccc ccacctacac cctcatctac 1140
cacaaaactt ccccaaggac tttcttttgc caagcgccta cagcagctca tagaggtctt 1200
gaaggggaag acctattccg acggaaagaa ccattatgac atggagacca cagagctccc 1260
agaactgacc tcgacctcct gatctgacct ggttctcaac agacccagtg agcccttcac 1320
tcctgagaaa aaggaaagat gaaataaata aataaacata tatatataga tataaaaaaa 1380
aaaaaaaa 1388
<210>183
<211>421
<212>PRT
<213>Homo sapiens
<220>
<223>Acrosin
<400>183
Met Val Glu Met Leu Pro Thr Ala Ile Leu Leu Val Leu Ala Val Ser
1 5 10 15
Val Val Ala Lys Asp Asn Ala Thr Cys Asp Gly Pro Cys Gly Leu Arg
20 25 30
Phe Arg Gln Asn Pro Gln Gly Gly Val Arg Ile Val Gly Gly Lys Ala
35 40 45
Ala Gln His Gly Ala Trp Pro Trp Met Val Ser Leu Gln Ile Phe Thr
50 55 60
Tyr Asn Ser His Arg Tyr His Thr Cys Gly Gly Ser Leu Leu Asn Ser
65 70 75 80
Arg Trp Val Leu Thr Ala Ala His Cys Phe Val Gly Lys Asn Asn Val
85 90 95
His Asp Trp Arg Leu ValPhe Gly Ala Lys Glu Ile Thr Tyr Gly Asn
100 105 110
Asn Lys Pro Val Lys Ala Pro Val Gln Glu Arg Tyr Val Glu Lys Ile
115 120 125
Ile Ile His Glu Lys Tyr Asn Ser Ala Thr Glu Gly Asn Asp Ile Ala
130 135 140
Leu ValGlu Ile Thr Pro Pro Ile Ser Cys Gly Arg Phe Ile Gly Pro
145 150 155 160
Gly Cys Leu Pro His Leu Lys Ala Gly Leu Pro Arg Gly Ser Gln Ser
165 170 175
Cys Trp Val Ala Gly Trp Gly Tyr Ile Glu Glu Lys Ala Pro Arg Pro
180 185 190
Ser Ser Ile Leu Met Glu Ala Arg Val Asp Leu Ile Asp Leu Asp Leu
195 200 205
Cys Asn Ser Thr Gln Trp Tyr Asn Gly Arg Val Gln Pro Thr Asn Val
210 215 220
Cys Ala Gly Tyr Pro Val Gly Lys Ile Asp Thr Cys Gln Gly Asp Ser
225 230 235 240
Gly Gly Pro Leu Met Cys Lys Asp Ser Lys Glu Ser Ala Tyr Val Val
245 250 255
ValGly Ile Thr Ser Trp Gly Val Gly Cys Ala Arg Ala Lys Arg Pro
260 265 270
Gly Ile Tyr Thr Ala Thr Trp Pro Tyr Leu Asn Trp Ile Ala Ser Lys
275 280 285
Ile Gly Ser Asn Ala Leu Arg Met Ile Gln Ser Ala Thr Pro Pro Pro
290 295 300
Pro Thr Thr Arg Pro Pro Pro Ile Arg Pro Pro Phe Ser His Pro Ile
305 310 315 320
Ser Ala His Leu Pro Trp Tyr Phe Gln Pro Pro Pro Arg Pro Leu Pro
325 330 335
Pro Arg Pro Pro Ala Ala Gln Pro Arg Pro Pro Pro Ser Pro Pro Pro
340 345 350
Pro Pro Pro Pro Pro Ala Ser Pro Leu Pro Pro Pro Pro Pro Pro Pro
355 360 365
Pro Pro Thr Pro Ser Ser Thr Thr Lys Leu Pro Gln Gly Leu Ser Phe
370 375 380
Ala Lys Arg Leu Gln Gln Leu Ile Glu Val Leu Lys Gly Lys Thr Tyr
385 390 395 400
Ser Asp Gly Lys Asn His Tyr Asp Met Glu Thr Thr Glu Leu Pro Glu
405 410 415
Leu Thr Ser Thr Ser
420
<210>184
<211>2363
<212>DNA
<213>Homo sapiens
<220>
<223>Hepsin transcript variant 1
<400>184
tcgagcccgc tttccaggga ccctacctga gggcccacag gtgaggcagc ctggcctagc 60
aggccccacg ccaccgcctc tgcctccagg ccgcccgctg ctgcggggcc accatgctcc 120
tgcccaggcc tggagactga cccgaccccg gcactacctc gaggctccgc ccccacctgc 180
tggaccccag ggtaaggaca agggccccca gactcacagt tccagccctg aggacagggg 240
ttccctcatc cccccaccca gcctaatgcc cacctcctaa tagaggggtt cctggggacc 300
tgaagagggg gcactatgac gtctccccaa gcacctaggt gttctgtcct gctcttcctt 360
cagactcagc cgttggaccc cagtcctttc ctccccagac ccaggagttc cagccctcag 420
gcccctcctc cctcatacta gggagtcctg gcccccaaat tcctcctttc ccaagactta 480
tgatttcagg tcctcagctg tctcctccct caaaccggga tcctcagtcc cctgctccac 540
caggctcagg catgggggtc cccatccctg caaatccagg cgtccccccg ctgctggtca 600
gacactgacc ccatccttga acccagccca atctgcgtcc gtgatcacgg cgtgctctgg 660
ccaaggccca gtccctacag cctgcctgga tggacgcctg ggactggggg cgccaggact 720
gggctgggct gggctccccc aggccctgcc tccccgtcca tctcctcaca ggtcccaccc 780
tggcccagga ggtcagccag ggaatcatta acaagaggca gtgacatggc gcagaaggag 840
ggtggccgga ctgtgccatg ctgctccaga cccaaggtgg cagctctcac tgcggggacc 900
ctgctacttc tgacagccat cggggcggca tcctgggcca ttgtggctgt tctcctcagg 960
agtgaccagg agccgctgta cccagtgcag gtcagctctg cggacgctcg gctcatggtc 1020
tttgacaaga cggaagggac gtggcggctg ctgtgctcct cgcgctccaa cgccagggta 1080
gccggactca gctgcgagga gatgggcttc ctcagggcac tgacccactc cgagctggac 1140
gtgcgaacgg cgggcgccaa tggcacgtcg ggcttcttct gtgtggacga ggggaggctg 1200
ccccacaccc agaggctgct ggaggtcatc tccgtgtgtg attgccccag aggccgtttc 1260
ttggccgcca tctgccaaga ctgtggccgc aggaagctgc ccgtggaccg catcgtggga 1320
ggccgggaca ccagcttggg ccggtggccg tggcaagtca gccttcgcta tgatggagca 1380
cacctctgtg ggggatccct gctctccggg gactgggtgc tgacagccgc ccactgcttc 1440
ccggagcgga accgggtcct gtcccgatgg cgagtgtttg ccggtgccgt ggcccaggcc 1500
tctccccacg gtctgcagct gggggtgcag gctgtggtct accacggggg ctatcttccc 1560
tttcgggacc ccaacagcga ggagaacagc aacgatattg ccctggtcca cctctccagt 1620
cccctgcccc tcacagaata catccagcct gtgtgcctcc cagctgccgg ccaggccctg 1680
gtggatggca agatctgtac cgtgacgggc tggggcaaca cgcagtacta tggccaacag 1740
gccggggtac tccaggaggc tcgagtcccc ataatcagca atgatgtctg caatggcgct 1800
gacttctatg gaaaccagat caagcccaag atgttctgtg ctggctaccc cgagggtggc 1860
attgatgcct gccagggcga cagcggtggt ccctttgtgt gtgaggacag catctctcgg 1920
acgccacgtt ggcggctgtg tggcattgtg agttggggca ctggctgtgc cctggcccag 1980
aagccaggcg tctacaccaa agtcagtgac ttccgggagt ggatcttcca ggccataaag 2040
actcactccg aagccagcgg catggtgacc cagctctgac cggtggcttc tcgctgcgca 2100
gcctccaggg cccgaggtga tcccggtggt gggatccacg ctgggccgag gatgggacgt 2160
ttttcttctt gggcccggtc cacaggtcca aggacaccct ccctccaggg tcctctcttc 2220
cacagtggcg ggcccactca gccccgagac cacccaacct caccctcctg acccccatgt 2280
aaatattgtt ctgctgtctg ggactcctgt ctaggtgccc ctgatgatgg gatgctcttt 2340
aaataataaa gatggttttg att 2363
<210>185
<211>417
<212>PRT
<213>Homo sapiens
<220>
<223>Hepsin
<400>185
Met Ala Gln Lys Glu Gly Gly Arg Thr Val Pro Cys Cys Ser Arg Pro
1 5 10 15
Lys Val Ala Ala Leu Thr Ala Gly Thr Leu Leu Leu Leu Thr Ala Ile
20 25 30
Gly Ala Ala Ser Trp Ala Ile Val Ala Val Leu Leu Arg Ser Asp Gln
35 40 45
Glu Pro Leu Tyr Pro Val Gln Val Ser Ser Ala Asp Ala Arg Leu Met
50 55 60
Val Phe Asp Lys Thr Glu Gly Thr Trp Arg Leu Leu Cys Ser Ser Arg
65 70 75 80
Ser Asn Ala Arg Val Ala Gly Leu Ser Cys Glu Glu Met Gly Phe Leu
85 90 95
Arg Ala Leu Thr His Ser Glu Leu Asp Val Arg Thr Ala Gly Ala Asn
100 105 110
Gly Thr Ser Gly Phe Phe Cys Val Asp Glu Gly Arg Leu Pro His Thr
115 120 125
Gln Arg Leu Leu Glu Val Ile Ser Val Cys Asp Cys Pro Arg Gly Arg
130 135 140
Phe Leu Ala Ala Ile Cys Gln Asp Cys Gly Arg Arg Lys Leu Pro Val
145 150 155 160
Asp Arg Ile Val Gly Gly Arg Asp Thr Ser Leu Gly Arg Trp Pro Trp
165 170 175
Gln ValSer Leu Arg Tyr Asp Gly Ala His Leu Cys Gly Gly Ser Leu
180 185 190
Leu Ser Gly Asp Trp Val Leu Thr Ala Ala His Cys Phe Pro Glu Arg
195 200 205
Asn Arg Val Leu Ser Arg Trp Arg Val Phe Ala Gly Ala Val Ala Gln
210 215 220
Ala Ser Pro His Gly Leu Gln Leu Gly Val Gln Ala Val Val Tyr His
225 230 235 240
Gly Gly Tyr Leu Pro Phe Arg Asp Pro Asn Ser Glu Glu Asn Ser Asn
245 250 255
Asp Ile Ala Leu Val His Leu Ser Ser Pro Leu Pro Leu Thr Glu Tyr
260 265 270
Ile Gln Pro Val Cys Leu Pro Ala Ala Gly Gln Ala Leu Val Asp Gly
275 280 285
Lys Ile Cys Thr ValThr Gly Trp Gly Asn Thr Gln Tyr Tyr Gly Gln
290 295 300
Gln Ala Gly Val Leu Gln Glu Ala Arg Val Pro Ile Ile Ser Asn Asp
305 310 315 320
Val Cys Asn Gly Ala Asp Phe Tyr Gly Asn Gln Ile Lys Pro Lys Met
325 330 335
Phe Cys Ala Gly Tyr Pro Glu Gly Gly Ile Asp Ala Cys Gln Gly Asp
340 345 350
Ser Gly Gly Pro Phe ValCys Glu Asp Ser Ile Ser Arg Thr Pro Arg
355 360 365
Trp Arg Leu Cys Gly Ile Val Ser Trp Gly Thr Gly Cys Ala Leu Ala
370 375 380
Gln Lys Pro Gly Val Tyr Thr Lys Val Ser Asp Phe Arg Glu Trp Ile
385 390 395 400
Phe Gln Ala Ile Lys Thr His Ser Glu Ala Ser Gly Met Val Thr Gln
405 410 415
Leu
<210>186
<211>1783
<212>DNA
<213>Homo sapiens
<220>
<223>Hepsin transcript variant 2
<400>186
tcgagcccgc tttccaggga ccctacctga gggcccacag gtgaggcagc ctggcctagc 60
aggccccacg ccaccgcctc tgcctccagg ccgcccgctg ctgcggggcc accatgctcc 120
tgcccaggcc tggagactga cccgaccccg gcactacctc gaggctccgc ccccacctgc 180
tggaccccag ggtcccaccc tggcccagga ggtcagccag ggaatcatta acaagaggca 240
gtgacatggc gcagaaggag ggtggccgga ctgtgccatg ctgctccaga cccaaggtgg 300
cagctctcac tgcggggacc ctgctacttc tgacagccat cggggcggca tcctgggcca 360
ttgtggctgt tctcctcagg agtgaccagg agccgctgta cccagtgcag gtcagctctg 420
cggacgctcg gctcatggtc tttgacaaga cggaagggac gtggcggctg ctgtgctcct 480
cgcgctccaa cgccagggta gccggactca gctgcgagga gatgggcttc ctcagggcac 540
tgacccactc cgagctggac gtgcgaacgg cgggcgccaa tggcacgtcg ggcttcttct 600
gtgtggacga ggggaggctg ccccacaccc agaggctgct ggaggtcatc tccgtgtgtg 660
attgccccag aggccgtttc ttggccgcca tctgccaaga ctgtggccgc aggaagctgc 720
ccgtggaccg catcgtggga ggccgggaca ccagcttggg ccggtggccg tggcaagtca 780
gccttcgcta tgatggagca cacctctgtg ggggatccct gctctccggg gactgggtgc 840
tgacagccgc ccactgcttc ccggagcgga accgggtcct gtcccgatgg cgagtgtttg 900
ccggtgccgt ggcccaggcc tctccccacg gtctgcagct gggggtgcag gctgtggtct 960
accacggggg ctatcttccc tttcgggacc ccaacagcga ggagaacagc aacgatattg 1020
ccctggtcca cctctccagt cccctgcccc tcacagaata catccagcct gtgtgcctcc 1080
cagctgccgg ccaggccctg gtggatggca agatctgtac cgtgacgggc tggggcaaca 1140
cgcagtacta tggccaacag gccggggtac tccaggaggc tcgagtcccc ataatcagca 1200
atgatgtctg caatggcgct gacttctatg gaaaccagat caagcccaag atgttctgtg 1260
ctggctaccc cgagggtggc attgatgcct gccagggcga cagcggtggt ccctttgtgt 1320
gtgaggacag catctctcgg acgccacgtt ggcggctgtg tggcattgtg agttggggca 1380
ctggctgtgc cctggcccag aagccaggcg tctacaccaa agtcagtgac ttccgggagt 1440
ggatcttcca ggccataaag actcactccg aagccagcgg catggtgacc cagctctgac 1500
cggtggcttc tcgctgcgca gcctccaggg cccgaggtga tcccggtggt gggatccacg 1560
ctgggccgag gatgggacgt ttttcttctt gggcccggtc cacaggtcca aggacaccct 1620
ccctccaggg tcctctcttc cacagtggcg ggcccactca gccccgagac cacccaacct 1680
caccctcctg acccccatgt aaatattgtt ctgctgtctg ggactcctgt ctaggtgccc 1740
ctgatgatgg gatgctcttt aaataataaa gatggttttg att 1783
<210>187
<211>417
<212>PRT
<213>Homo sapiens
<220>
<223>Hepsin
<400>187
Met Ala Gln Lys Glu Gly Gly Arg Thr Val Pro Cys Cys Ser Arg Pro
1 5 10 15
Lys Val Ala Ala Leu Thr Ala Gly Thr Leu Leu Leu Leu Thr Ala Ile
20 25 30
Gly Ala Ala Ser Trp Ala Ile Val Ala Val Leu Leu Arg Ser Asp Gln
35 40 45
Glu Pro Leu Tyr Pro Val Gln Val Ser Ser Ala Asp Ala Arg Leu Met
50 55 60
Val Phe Asp Lys Thr Glu Gly Thr Trp Arg Leu Leu Cys Ser Ser Arg
65 70 75 80
Ser Asn Ala Arg Val Ala Gly Leu Ser Cys Glu Glu Met Gly Phe Leu
85 90 95
Arg Ala Leu Thr His Ser Glu Leu Asp Val Arg Thr Ala Gly Ala Asn
100 105 110
Gly Thr Ser Gly Phe Phe Cys Val Asp Glu Gly Arg Leu Pro His Thr
115 120 125
Gln Arg Leu Leu Glu Val Ile Ser Val Cys Asp Cys Pro Arg Gly Arg
130 135 140
Phe Leu Ala Ala Ile Cys Gln Asp Cys Gly Arg Arg Lys Leu Pro Val
145 150 155 160
Asp Arg Ile Val Gly Gly Arg Asp Thr Ser Leu Gly Arg Trp Pro Trp
165 170 175
Gln Val Ser Leu Arg Tyr Asp Gly Ala His Leu Cys Gly Gly Ser Leu
180 185 190
Leu Ser Gly Asp Trp Val Leu Thr Ala Ala His Cys Phe Pro Glu Arg
195 200 205
Asn Arg Val Leu Ser Arg Trp Arg Val Phe Ala Gly Ala Val Ala Gln
210 215 220
Ala Ser Pro His Gly Leu Gln Leu Gly Val Gln Ala Val Val Tyr His
225 230 235 240
Gly Gly Tyr Leu Pro Phe Arg Asp Pro Asn Ser Glu Glu Asn Ser Asn
245 250 255
Asp Ile Ala Leu Val His Leu Ser Ser Pro Leu Pro Leu Thr Glu Tyr
260 265 270
Ile Gln Pro Val Cys Leu Pro Ala Ala Gly Gln Ala Leu Val Asp Gly
275 280 285
Lys Ile Cys Thr Val Thr Gly Trp Gly Asn Thr Gln Tyr Tyr Gly Gln
290 295 300
Gln Ala Gly Val Leu Gln Glu Ala Arg Val Pro Ile Ile Ser Asn Asp
305 310 315 320
Val Cys Asn Gly Ala Asp Phe Tyr Gly Asn Gln Ile Lys Pro Lys Met
325 330 335
Phe Cys Ala Gly Tyr Pro Glu Gly Gly Ile Asp Ala Cys Gln Gly Asp
340 345 350
Ser Gly Gly Pro Phe Val Cys Glu Asp Ser Ile Ser Arg Thr Pro Arg
355 360 365
Trp Arg Leu Cys Gly Ile Val Ser Trp Gly Thr Gly Cys Ala Leu Ala
370 375 380
Gln Lys Pro Gly ValTyr Thr Lys Val Ser Asp Phe Arg Glu Trp Ile
385 390 395 400
Phe Gln Ala Ile Lys Thr His Ser Glu Ala Ser Gly Met Val Thr Gln
405 410 415
Leu
<210>188
<211>2038
<212>DNA
<213>Homo sapiens
<220>
<223>hepatocyte growth factor activator
<400>188
gccatggggc gctgggcctg ggtccccagc ccctggcccc caccggggct gggccccttc 60
ctcctcctcc tcctgctgct gctgctgctg ccacgggggt tccagcccca gcctggcggg 120
aaccgtacgg agtccccaga acctaatgcc acagcgaccc ctgcgatccc cactatcctg 180
gtgacctctg tgacctctga gaccccagca acaagtgctc cagaggcaga gggaccccaa 240
agtggggggc tcccgccccc gcccagggca gttccctcga gcagtagccc ccaggcccaa 300
gcactcaccg aggacgggag gccctgcagg ttccccttcc gctacggggg ccgcatgctg 360
catgcctgca cttcggaggg cagtgcacac aggaagtggt gtgccacaac tcacaactac 420
gaccgggaca gggcctgggg ctactgtgtg gaggccaccc cgcctccagg gggcccagct 480
gccctggatc cctgtgcctc cggcccctgc ctcaatggag gctcctgctc caatacccag 540
gacccccagt cctatcactg cagctgcccc cgggccttca ccggcaagga ctgcggcaca 600
gagaaatgct ttgatgagac ccgctacgag tacctggagg ggggcgaccg ctgggcccgc 660
gtgcgccagg gccacgtgga acagtgcgag tgcttcgggg gccggacctg gtgcgaaggc 720
acccgacata cagcttgtct gagcagccct tgcctgaacg ggggcacctg ccacctgatc 780
gtggccaccg ggaccaccgt gtgtgcctgc ccaccaggct tcgctggacg gctctgcaac 840
atcgagcctg atgagcgctg cttcttgggg aacggcactg ggtaccgtgg cgtggccagc 900
acctcagcct cgggcctcag ctgcctggcc tggaactccg atctgctcta ccaggagctg 960
cacgtggact ccgtgggcgc cgcggccctg ctgggcctgg gcccccatgc ctactgccgg 1020
aatccggaca atgacgagag gccctggtgc tacgtggtga aggacagcgc gctctcctgg 1080
gagtactgcc gcctggaggc ctgcgaatcc ctcaccagag tccaactgtc accggatctc 1140
ctggcgaccc tgcctgagcc agcctccccg gggcgccagg cctgcggcag gaggcacaag 1200
aagaggacgt tcctgcggcc acgtatcatc ggcggctcct cctcgctgcc cggctcgcac 1260
ccctggctgg ccgccatcta catcggggac agcttctgcg ccgggagcct ggtccacacc 1320
tgctgggtgg tgtcggccgc ccactgcttc tcccacagcc cccccaggga cagcgtctcc 1380
gtggtgctgg gccagcactt cttcaaccgc acgacggacg tgacgcagac cttcggcatc 1440
gagaagtaca tcccgtacac cctgtactcg gtgttcaacc ccagcgacca cgacctcgtc 1500
ctgatccggc tgaagaagaa aggggaccgc tgtgccacac gctcgcagtt cgtgcagccc 1560
atctgcctgc ccgagcccgg cagcaccttc cccgcaggac acaagtgcca gattgcgggc 1620
tggggccact tggatgagaa cgtgagcggc tactccagct ccctgcggga ggccctggtc 1680
cccctggtcg ccgaccacaa gtgcagcagc cctgaggtct acggcgccga catcagcccc 1740
aacatgctct gtgccggcta cttcgactgc aagtccgacg cctgccaggg ggactcaggg 1800
gggcccctgg cctgcgagaa gaacggcgtg gcttacctct acggcatcat cagctggggt 1860
gacggctgcg ggcggctcca caagccgggg gtctacaccc gcgtggccaa ctatgtggac 1920
tggatcaacg accggatacg gcctcccagg cggcttgtgg ctccctcctg accctccagc 1980
gggacaccct ggttcccacc attccctgcc ttgctgacaa taaagatatt tccaagaa 2038
<210>189
<211>655
<212>PRT
<213>Homo sapiens
<220>
<223>hepatocyte growth factor activator
<400>189
Met Gly Arg Trp Ala Trp Val Pro Ser Pro Trp Pro Pro Pro Gly Leu
1 5 10 15
Gly Pro Phe Leu Leu Leu Leu Leu Leu Leu Leu Leu Leu Pro Arg Gly
20 25 30
Phe Gln Pro Gln Pro Gly Gly Asn Arg Thr Glu Ser Pro Glu Pro Asn
35 40 45
Ala Thr Ala Thr Pro Ala Ile Pro Thr Ile Leu Val Thr Ser Val Thr
50 55 60
Ser Glu Thr Pro Ala Thr Ser Ala Pro Glu Ala Glu Gly Pro Gln Ser
65 70 75 80
Gly Gly Leu Pro Pro Pro Pro Arg Ala Val Pro Ser Ser Ser Ser Pro
85 90 95
Gln Ala Gln Ala Leu Thr Glu Asp Gly Arg Pro Cys Arg Phe Pro Phe
100 105 110
Arg Tyr Gly Gly Arg Met Leu His Ala Cys Thr Ser Glu Gly Ser Ala
115 120 125
His Arg Lys Trp Cys Ala Thr Thr His Asn Tyr Asp Arg Asp Arg Ala
130 135 140
Trp Gly Tyr Cys Val Glu Ala Thr Pro Pro Pro Gly Gly Pro Ala Ala
145 150 155 160
Leu Asp Pro Cys Ala Ser Gly Pro Cys Leu Asn Gly Gly Ser Cys Ser
165 170 175
Asn Thr Gln Asp Pro Gln Ser Tyr His Cys Ser Cys Pro Arg Ala Phe
180 185 190
Thr Gly Lys Asp Cys Gly Thr Glu Lys Cys Phe Asp Glu Thr Arg Tyr
195 200 205
Glu Tyr Leu Glu Gly Gly Asp Arg Trp Ala Arg Val Arg Gln Gly His
210 215 220
Val Glu Gln Cys Glu Cys Phe Gly Gly Arg Thr Trp Cys Glu Gly Thr
225 230 235 240
Arg His Thr Ala Cys Leu Ser Ser Pro Cys Leu Asn Gly Gly Thr Cys
245 250 255
His Leu Ile Val Ala Thr Gly Thr Thr Val Cys Ala Cys Pro Pro Gly
260 265 270
Phe Ala Gly Arg Leu Cys Asn Ile Glu Pro Asp Glu Arg Cys Phe Leu
275 280 285
Gly Asn Gly Thr Gly Tyr Arg Gly Val Ala Ser Thr Ser Ala Ser Gly
290 295 300
Leu Ser Cys Leu Ala Trp Asn Ser Asp Leu Leu Tyr Gln Glu Leu His
305 310 315 320
Val Asp Ser Val Gly Ala Ala Ala Leu Leu Gly Leu Gly Pro His Ala
325 330 335
Tyr Cys Arg Asn Pro Asp Asn Asp Glu Arg Pro Trp Cys Tyr Val Val
340 345 350
Lys Asp Ser Ala Leu Ser Trp Glu Tyr Cys Arg Leu Glu Ala Cys Glu
355 360 365
Ser Leu Thr Arg Val Gln Leu Ser Pro Asp Leu Leu Ala Thr Leu Pro
370 375 380
Glu Pro Ala Ser Pro Gly Arg Gln Ala Cys Gly Arg Arg His Lys Lys
385 390 395 400
Arg Thr Phe Leu Arg Pro Arg Ile Ile Gly Gly Ser Ser Ser Leu Pro
405 410 415
Gly Ser His Pro Trp Leu Ala Ala Ile Tyr Ile Gly Asp Ser Phe Cys
420 425 430
Ala Gly Ser Leu Val His Thr Cys Trp Val Val Ser Ala Ala His Cys
435 440 445
Phe Ser His Ser Pro Pro Arg Asp Ser Val Ser Val Val Leu Gly Gln
450 455 460
His Phe Phe Asn Arg Thr Thr Asp Val Thr Gln Thr Phe Gly Ile Glu
465 470 475 480
Lys Tyr Ile Pro Tyr Thr Leu Tyr Ser Val Phe Asn Pro Ser Asp His
485 490 495
Asp Leu Val Leu Ile Arg Leu Lys Lys Lys Gly Asp Arg Cys Ala Thr
500 505 510
Arg Ser Gln Phe Val Gln Pro Ile Cys Leu Pro Glu Pro Gly Ser Thr
515 520 525
Phe Pro Ala Gly His Lys Cys Gln Ile Ala Gly Trp Gly His Leu Asp
530 535 540
Glu Asn Val Ser Gly Tyr Ser Ser Ser Leu Arg Glu Ala Leu Val Pro
545 550 555 560
Leu ValAla Asp His Lys Cys Ser Ser Pro Glu Val Tyr Gly Ala Asp
565 570 575
Ile Ser Pro Asn Met Leu Cys Ala Gly Tyr Phe Asp Cys Lys Ser Asp
580 585 590
Ala Cys Gln Gly Asp Ser Gly Gly Pro Leu Ala Cys Glu Lys Asn Gly
595 600 605
Val Ala Tyr Leu Tyr Gly Ile Ile Ser Trp Gly Asp Gly Cys Gly Arg
610 615 620
Leu His Lys Pro Gly Val Tyr Thr Arg Val Ala Asn Tyr Val Asp Trp
625 630 635 640
Ile Asn Asp Arg Ile Arg Pro Pro Arg Arg Leu Val Ala Pro Ser
645 650 655
<210>190
<211>2360
<212>DNA
<213>Homo sapiens
<220>
<223>u-plasminogen activator (uPA)
<400>190
acagtgcgga gaccgcagcc ccggagcccg ggccagggtc cacctgtccc cgcagcgccg 60
gctcgcgccc tcctgccgca gccaccgagc cgccgtctag cgccccgacc tcgccaccat 120
gagagccctg ctggcgcgcc tgcttctctg cgtcctggtc gtgagcgact ccaaaggcag 180
caatgaactt catcaagttc catcgaactg tgactgtcta aatggaggaa catgtgtgtc 240
caacaagtac ttctccaaca ttcactggtg caactgccca aagaaattcg gagggcagca 300
ctgtgaaata gataagtcaa aaacctgcta tgaggggaat ggtcactttt accgaggaaa 360
ggccagcact gacaccatgg gccggccctg cctgccctgg aactctgcca ctgtccttca 420
gcaaacgtac catgcccaca gatctgatgc tcttcagctg ggcctgggga aacataatta 480
ctgcaggaac ccagacaacc ggaggcgacc ctggtgctat gtgcaggtgg gcctaaagcc 540
gcttgtccaa gagtgcatgg tgcatgactg cgcagatgga aaaaagccct cctctcctcc 600
agaagaatta aaatttcagt gtggccaaaa gactctgagg ccccgcttta agattattgg 660
gggagaattc accaccatcg agaaccagcc ctggtttgcg gccatctaca ggaggcaccg 720
ggggggctct gtcacctacg tgtgtggagg cagcctcatc agcccttgct gggtgatcag 780
cgccacacac tgcttcattg attacccaaa gaaggaggac tacatcgtct acctgggtcg 840
ctcaaggctt aactccaaca cgcaagggga gatgaagttt gaggtggaaa acctcatcct 900
acacaaggac tacagcgctg acacgcttgc tcaccacaac gacattgcct tgctgaagat 960
ccgttccaag gagggcaggt gtgcgcagcc atcccggact atacagacca tctgcctgcc 1020
ctcgatgtat aacgatcccc agtttggcac aagctgtgag atcactggct ttggaaaaga 1080
gaattctacc gactatctct atccggagca gctgaaaatg actgttgtga agctgatttc 1140
ccaccgggag tgtcagcagc cccactacta cggctctgaa gtcaccacca aaatgctgtg 1200
tgctgctgac ccacagtgga aaacagattc ctgccaggga gactcagggg gacccctcgt 1260
ctgttccctc caaggccgca tgactttgac tggaattgtg agctggggcc gtggatgtgc 1320
cctgaaggac aagccaggcg tctacacgag agtctcacac ttcttaccct ggatccgcag 1380
tcacaccaag gaagagaatg gcctggccct ctgagggtcc ccagggagga aacgggcacc 1440
acccgctttc ttgctggttg tcatttttgc agtagagtca tctccatcag ctgtaagaag 1500
agactgggaa gataggctct gcacagatgg atttgcctgt gccacccacc agggcgaacg 1560
acaatagctt taccctcagg cataggcctg ggtgctggct gcccagaccc ctctggccag 1620
gatggagggg tggtcctgac tcaacatgtt actgaccagc aacttgtctt tttctggact 1680
gaagcctgca ggagttaaaa agggcagggc atctcctgtg catgggtgaa gggagagcca 1740
gctcccccga cggtgggcat ttgtgaggcc catggttgag aaatgaataa tttcccaatt 1800
aggaagtgta acagctgagg tctcttgagg gagcttagcc aatgtgggag cagcggtttg 1860
gggagcagag acactaacga cttcagggca gggctctgat attccatgaa tgtatcagga 1920
aatatatatg tgtgtgtatg tttgcacact tgtgtgtggg ctgtgagtgt aagtgtgagt 1980
aagagctggt gtctgattgt taagtctaaa tatttcctta aactgtgtgg actgtgatgc 2040
cacacagagt ggtctttctg gagaggttat aggtcactcc tggggcctct tgggtccccc 2100
acgtgacagt gcctgggaat gtattattct gcagcatgac ctgtgaccag cactgtctca 2160
gtttcacttt cacatagatg tccctttctt ggccagttat cccttccttt tagcctagtt 2220
catccaatcc tcactgggtg gggtgaggac cactcctgta cactgaatat ttatatttca 2280
ctatttttat ttatattttt gtaattttaa ataaaagtga tcaataaaat gtgatttttc 2340
tgatgaaaaa aaaaaaaaaa 2360
<210>191
<211>431
<212>PRT
<213>Homo sapiens
<220>
<223>u-plasminogen activator (uPA)
<400>191
Met Arg Ala Leu Leu Ala Arg Leu Leu Leu Cys Val Leu Val Val Ser
1 5 10 15
Asp Ser Lys Gly Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp
20 25 30
Cys Leu Asn Gly Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile
35 40 45
His Trp Cys Asn Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile
50 55 60
Asp Lys Ser Lys Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly
65 70 75 80
Lys Ala Ser Thr Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser
85 90 95
Ala Thr Val Leu Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu
100 105 110
Gln Leu Gly Leu Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg
115 120 125
Arg Arg Pro Trp Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln
130 135 140
Glu Cys Met Val His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro
145 150 155 160
Pro Glu Glu Leu Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg
165 170 175
Phe Lys Ile Ile Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp
180 185 190
Phe Ala Ala Ile Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val
195 200 205
Cys Gly Gly Ser Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His
210 215 220
Cys Phe Ile Asp Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly
225 230 235 240
Arg Ser Arg Leu Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val
245 250 255
Glu Asn Leu Ile Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His
260 265 270
His Asn Asp Ile Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys
275 280 285
Ala Gln Pro Ser Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr
290 295 300
Asn Asp Pro Gln Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys
305 310 315 320
Glu Asn Ser Thr Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val
325 330 335
Val Lys Leu Ile Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly
340 345 350
Ser Glu Val Thr Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys
355 360 365
Thr Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu
370 375 380
Gln Gly Arg Met Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys
385 390 395 400
Ala Leu Lys Asp Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu
405 410 415
Pro Trp Ile Arg Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
420 425 430
<210>192
<211>2653
<212>DNA
<213>Homo sapiens
<220>
<223>t-plasminogen activator (tPA)transcript variant 1
<400>192
atggccctgt ccactgagca tcctcccgcc acacagaaac ccgcccagcc ggggccaccg 60
accccacccc ctgcctggaa acttaaggag gccggagctg tggggagctc agagctgaga 120
tcctacagga gtccagggct ggagagaaaa cctctgcgag gaaagggaag gagcaagccg 180
tgaatttaag ggacgctgtg aagcaatcat ggatgcaatg aagagagggc tctgctgtgt 240
gctgctgctg tgtggagcag tcttcgtttc gcccagccag gaaatccatg cccgattcag 300
aagaggagcc agatcttacc aagtgatctg cagagatgaa aaaacgcaga tgatatacca 360
gcaacatcag tcatggctgc gccctgtgct cagaagcaac cgggtggaat attgctggtg 420
caacagtggc agggcacagt gccactcagt gcctgtcaaa agttgcagcg agccaaggtg 480
tttcaacggg ggcacctgcc agcaggccct gtacttctca gatttcgtgt gccagtgccc 540
cgaaggattt gctgggaagt gctgtgaaat agataccagg gccacgtgct acgaggacca 600
gggcatcagc tacaggggca cgtggagcac agcggagagt ggcgccgagt gcaccaactg 660
gaacagcagc gcgttggccc agaagcccta cagcgggcgg aggccagacg ccatcaggct 720
gggcctgggg aaccacaact actgcagaaa cccagatcga gactcaaagc cctggtgcta 780
cgtctttaag gcggggaagt acagctcaga gttctgcagc acccctgcct gctctgaggg 840
aaacagtgac tgctactttg ggaatgggtc agcctaccgt ggcacgcaca gcctcaccga 900
gtcgggtgcc tcctgcctcc cgtggaattc catgatcctg ataggcaagg tttacacagc 960
acagaacccc agtgcccagg cactgggcct gggcaaacat aattactgcc ggaatcctga 1020
tggggatgcc aagccctggt gccacgtgct gaagaaccgc aggctgacgt gggagtactg 1080
tgatgtgccc tcctgctcca cctgcggcct gagacagtac agccagcctc agtttcgcat 1140
caaaggaggg ctcttcgccg acatcgcctc ccacccctgg caggctgcca tctttgccaa 1200
gcacaggagg tcgcccggag agcggttcct gtgcgggggc atactcatca gctcctgctg 1260
gattctctct gccgcccact gcttccagga gaggtttccg ccccaccacc tgacggtgat 1320
cttgggcaga acataccggg tggtccctgg cgaggaggag cagaaatttg aagtcgaaaa 1380
atacattgtc cataaggaat tcgatgatga cacttacgac aatgacattg cgctgctgca 1440
gctgaaatcg gattcgtccc gctgtgccca ggagagcagc gtggtccgca ctgtgtgcct 1500
tcccccggcg gacctgcagc tgccggactg gacggagtgt gagctctccg gctacggcaa 1560
gcatgaggcc ttgtctcctt tctattcgga gcggctgaag gaggctcatg tcagactgta 1620
cccatccagc cgctgcacat cacaacattt acttaacaga acagtcaccg acaacatgct 1680
gtgtgctgga gacactcgga gcggcgggcc ccaggcaaac ttgcacgacg cctgccaggg 1740
cgattcggga ggccccctgg tgtgtctgaa cgatggccgc atgactttgg tgggcatcat 1800
cagctggggc ctgggctgtg gacagaagga tgtcccgggt gtgtacacca aggttaccaa 1860
ctacctagac tggattcgtg acaacatgcg accgtgacca ggaacacccg actcctcaaa 1920
agcaaatgag atcccgcctc ttcttcttca gaagacactg caaaggcgca gtgcttctct 1980
acagacttct ccagacccac cacaccgcag aagcgggacg agaccctaca ggagagggaa 2040
gagtgcattt tcccagatac ttcccatttt ggaagttttc aggacttggt ctgatttcag 2100
gatactctgt cagatgggaa gacatgaatg cacactagcc tctccaggaa tgcctcctcc 2160
ctgggcagaa agtggccatg ccaccctgtt ttcagctaaa gcccaacctc ctgacctgtc 2220
accgtgagca gctttggaaa caggaccaca aaaatgaaag catgtctcaa tagtaaaaga 2280
taacaagatc tttcaggaaa gacggattgc attagaaata gacagtatat ttatagtcac 2340
aagagcccag cagggcctca aagttggggc aggctggctg gcccgtcatg ttcctcaaaa 2400
gcacccttga cgtcaagtct ccttcccctt tccccactcc ctggctctca gaaggtattc 2460
cttttgtgta cagtgtgtaa agtgtaaatc ctttttcttt ataaacttta gagtagcatg 2520
agagaattgt atcatttgaa caactaggct tcagcatatt tatagcaatc catgttagtt 2580
tttactttct gttgccacaa ccctgtttta tactgtactt aataaattca gatatatttt 2640
tcacagtttt tcc 2653
<210>193
<211>562
<212>PRT
<213>Homo sapiens
<220>
<223>t-plasminogen activator (tPA)isoform 1
<400>193
Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly
1 5 10 15
Ala Val Phe Val Ser Pro Ser Gln Glu Ile His Ala Arg Phe Arg Arg
20 25 30
Gly Ala Arg Ser Tyr Gln Val Ile Cys Arg Asp Glu Lys Thr Gln Met
35 40 45
Ile Tyr Gln Gln His Gln Ser Trp Leu Arg Pro Val Leu Arg Ser Asn
50 55 60
Arg Val Glu Tyr Cys Trp Cys Asn Ser Gly Arg Ala Gln Cys His Ser
65 70 75 80
Val Pro Val Lys Ser Cys Ser Glu Pro Arg Cys Phe Asn Gly Gly Thr
85 90 95
Cys Gln Gln Ala Leu Tyr Phe Ser Asp Phe Val Cys Gln Cys Pro Glu
100 105 110
Gly Phe Ala Gly Lys Cys Cys Glu Ile Asp Thr Arg Ala Thr Cys Tyr
115 120 125
Glu Asp Gln Gly Ile Ser Tyr Arg Gly Thr Trp Ser Thr Ala Glu Ser
130 135 140
Gly Ala Glu Cys Thr Asn Trp Asn Ser Ser Ala Leu Ala Gln Lys Pro
145 150 155 160
Tyr Ser Gly Arg Arg Pro Asp Ala Ile Arg Leu Gly Leu Gly Asn His
165 170 175
Asn Tyr Cys Arg Asn Pro Asp Arg Asp Ser Lys Pro Trp Cys Tyr Val
180 185 190
Phe Lys Ala Gly Lys Tyr Ser Ser Glu Phe Cys Ser Thr Pro Ala Cys
195 200 205
Ser Glu Gly Asn Ser Asp Cys Tyr Phe Gly Asn Gly Ser Ala Tyr Arg
210 215 220
Gly Thr His Ser Leu Thr Glu Ser Gly Ala Ser Cys Leu Pro Trp Asn
225 230 235 240
Ser Met Ile Leu Ile Gly Lys Val Tyr Thr Ala Gln Asn Pro Ser Ala
245 250 255
Gln Ala Leu Gly Leu Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Gly
260 265 270
Asp Ala Lys Pro Trp Cys His Val Leu Lys Asn Arg Arg Leu Thr Trp
275 280 285
Glu Tyr Cys Asp Val Pro Ser Cys Ser Thr Cys Gly Leu Arg Gln Tyr
290 295 300
Ser Gln Pro Gln Phe Arg Ile Lys Gly Gly Leu Phe Ala Asp Ile Ala
305 310 315 320
Ser His Pro Trp Gln Ala Ala Ile Phe Ala Lys His Arg Arg Ser Pro
325 330 335
Gly Glu Arg Phe Leu Cys Gly Gly Ile Leu Ile Ser Ser Cys Trp Ile
340 345 350
Leu Ser Ala Ala His Cys Phe Gln Glu Arg Phe Pro Pro His His Leu
355 360 365
Thr Val Ile Leu Gly Arg Thr Tyr Arg Val Val Pro Gly Glu Glu Glu
370 375 380
Gln Lys Phe Glu Val Glu Lys Tyr Ile Val His Lys Glu Phe Asp Asp
385 390 395 400
Asp Thr Tyr Asp Asn Asp Ile Ala Leu Leu Gln Leu Lys Ser Asp Ser
405 410 415
Ser Arg Cys Ala Gln Glu Ser Ser Val Val Arg Thr Val Cys Leu Pro
420 425 430
Pro Ala Asp Leu Gln Leu Pro Asp Trp Thr Glu Cys Glu Leu Ser Gly
435 440 445
Tyr Gly Lys His Glu Ala Leu Ser Pro Phe Tyr Ser Glu Arg Leu Lys
450 455 460
Glu Ala His Val Arg Leu Tyr Pro Ser Ser Arg Cys Thr Ser Gln His
465 470 475 480
Leu Leu Asn Arg Thr Val Thr Asp Asn Met Leu Cys Ala Gly Asp Thr
485 490 495
Arg Ser Gly Gly Pro Gln Ala Asn Leu His Asp Ala Cys Gln Gly Asp
500 505 510
Ser Gly Gly Pro Leu Val Cys Leu Asn Asp Gly Arg Met Thr Leu Val
515 520 525
Gly Ile Ile Ser Trp Gly Leu Gly Cys Gly Gln Lys Asp Val Pro Gly
530 535 540
Val Tyr Thr Lys Val Thr Asn Tyr Leu Asp Trp Ile Arg Asp Asn Met
545 550 555 560
Arg Pro
<210>194
<211>2160
<212>DNA
<213>Homo sapiens
<220>
<223>t-plasminogen activator (tPA)transcript variant 2
<400>194
atggccctgt ccactgagca tcctcccgcc acacagaaac ccgcccagcc ggggccaccg 60
accccacccc ctgcctggaa acttaaggag gccggagctg tggggagctc agagctgaga 120
tcctacagga gtccagggct ggagagaaaa cctctgcgag gaaagggaag gagcaagccg 180
tgaatttaag ggacgctgtg aagcaatcat ggatgcaatg aagagagggc tctgctgtgt 240
gctgctgctg tgtggagcag tcttcgtttc gcccagccag gaaatccatg cccgattcag 300
aagaggagcc agatcttacc aagtgatctg cagagatgaa aaaacgcaga tgatatacca 360
gcaacatcag tcatggctgc gccctgtgct cagaagcaac cgggtggaat attgctggtg 420
caacagtggc agggcacagt gccactcagt gcctgtcaaa agttgcagcg agccaaggtg 480
tttcaacggg ggcacctgcc agcaggccct gtacttctca gatttcgtgt gccagtgccc 540
cgaaggattt gctgggaagt gctgtgaaat agataccagg gccacgtgct acgaggacca 600
gggcatcagc tacaggggca cgtggagcac agcggagagt ggcgccgagt gcaccaactg 660
gaacagcagc gcgttggccc agaagcccta cagcgggcgg aggccagacg ccatcaggct 720
gggcctgggg aaccacaact actgcagaaa cccagatcga gactcaaagc cctggtgcta 780
cgtctttaag gcggggaagt acagctcaga gttctgcagc acccctgcct gctctgaggg 840
aaacagtgac tgctactttg ggaatgggtc agcctaccgt ggcacgcaca gcctcaccga 900
gtcgggtgcc tcctgcctcc cgtggaattc catgatcctg ataggcaagg tttacacagc 960
acagaacccc agtgcccagg cactgggcct gggcaaacat aattactgcc ggactggacg 1020
gagtgtgagc tctccggcta cggcaagcat gaggccttgt ctcctttcta ttcggagcgg 1080
ctgaaggagg ctcatgtcag actgtaccca tccagccgct gcacatcaca acatttactt 1140
aacagaacag tcaccgacaa catgctgtgt gctggagaca ctcggagcgg cgggccccag 1200
gcaaacttgc acgacgcctg ccagggcgat tcgggaggcc ccctggtgtg tctgaacgat 1260
ggccgcatga ctttggtggg catcatcagc tggggcctgg gctgtggaca gaaggatgtc 1320
ccgggtgtgt acaccaaggt taccaactac ctagactgga ttcgtgacaa catgcgaccg 1380
tgaccaggaa cacccgactc ctcaaaagca aatgagatcc cgcctcttct tcttcagaag 1440
acactgcaaa ggcgcagtgc ttctctacag acttctccag acccaccaca ccgcagaagc 1500
gggacgagac cctacaggag agggaagagt gcattttccc agatacttcc cattttggaa 1560
gttttcagga cttggtctga tttcaggata ctctgtcaga tgggaagaca tgaatgcaca 1620
ctagcctctc caggaatgcc tcctccctgg gcagaaagtg gccatgccac cctgttttca 1680
gctaaagccc aacctcctga cctgtcaccg tgagcagctt tggaaacagg accacaaaaa 1740
tgaaagcatg tctcaatagt aaaagataac aagatctttc aggaaagacg gattgcatta 1800
gaaatagaca gtatatttat agtcacaaga gcccagcagg gcctcaaagt tggggcaggc 1860
tggctggccc gtcatgttcc tcaaaagcac ccttgacgtc aagtctcctt cccctttccc 1920
cactccctgg ctctcagaag gtattccttt tgtgtacagt gtgtaaagtg taaatccttt 1980
ttctttataa actttagagt agcatgagag aattgtatca tttgaacaac taggcttcag 2040
catatttata gcaatccatg ttagttttta ctttctgttg ccacaaccct gttttatact 2100
gtacttaata aattcagata tatttttcac agtttttcca aaaaaaaaaa aaaaaaaaaa 2160
<210>195
<211>291
<212>PRT
<213>Homo sapiens
<220>
<223>t-plasminogen activator (tPA)isoform 2
<400>195
Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly
1 5 10 15
Ala Val Phe Val Ser Pro Ser Gln Glu Ile His Ala Arg Phe Arg Arg
20 25 30
Gly Ala Arg Ser Tyr Gln Val Ile Cys Arg Asp Glu Lys Thr Gln Met
35 40 45
Ile Tyr Gln Gln His Gln Ser Trp Leu Arg Pro Val Leu Arg Ser Asn
50 55 60
Arg Val Glu Tyr Cys Trp Cys Asn Ser Gly Arg Ala Gln Cys His Ser
65 70 75 80
Val Pro ValLys Ser Cys Ser Glu Pro Arg Cys Phe Asn Gly Gly Thr
85 90 95
Cys Gln Gln Ala Leu Tyr Phe Ser Asp Phe Val Cys Gln Cys Pro Glu
100 105 110
Gly Phe Ala Gly Lys Cys Cys Glu Ile Asp Thr Arg Ala Thr Cys Tyr
115 120 125
Glu Asp Gln Gly Ile Ser Tyr Arg Gly Thr Trp Ser Thr Ala Glu Ser
130 135 140
Gly Ala Glu Cys Thr Asn Trp Asn Ser Ser Ala Leu Ala Gln Lys Pro
145 150 155 160
Tyr Ser Gly Arg Arg Pro Asp Ala Ile Arg Leu Gly Leu Gly Asn His
165 170 175
Asn Tyr Cys Arg Asn Pro Asp Arg Asp Ser Lys Pro Trp Cys Tyr Val
180 185 190
Phe Lys Ala Gly Lys Tyr Ser Ser Glu Phe Cys Ser Thr Pro Ala Cys
195 200 205
Ser Glu Gly Asn Ser Asp Cys Tyr Phe Gly Asn Gly Ser Ala Tyr Arg
210 215 220
Gly Thr His Ser Leu Thr Glu Ser Gly Ala Ser Cys Leu Pro Trp Asn
225 230 235 240
Ser Met Ile Leu Ile Gly Lys Val Tyr Thr Ala Gln Asn Pro Ser Ala
245 250 255
Gln Ala Leu Gly Leu Gly Lys His Asn Tyr Cys Arg Thr Gly Arg Ser
260 265 270
Val Ser Ser Pro Ala Thr Ala Ser Met Arg Pro Cys Leu Leu Ser Ile
275 280 285
Arg Ser Gly
290
<210>196
<211>2534
<212>DNA
<213>Homo sapiens
<220>
<223>t-plasminogen activator (tPA)transcript variant 3
<400>196
atggccctgt ccactgagca tcctcccgcc acacagaaac ccgcccagcc ggggccaccg 60
accccacccc ctgcctggaa acttaaggag gccggagctg tggggagctc agagctgaga 120
tcctacagga gtccagggct ggagagaaaa cctctgcgag gaaagggaag gagcaagccg 180
tgaatttaag ggacgctgtg aagcaatcat ggatgcaatg aagagagggc tctgctgtgt 240
gctgctgctg tgtggagcag tcttcgtttc gcccagccag gaaatccatg cccgattcag 300
aagaggagcc agatcttacc aaggttgcag cgagccaagg tgtttcaacg ggggcacctg 360
ccagcaggcc ctgtacttct cagatttcgt gtgccagtgc cccgaaggat ttgctgggaa 420
gtgctgtgaa atagatacca gggccacgtg ctacgaggac cagggcatca gctacagggg 480
cacgtggagc acagcggaga gtggcgccga gtgcaccaac tggaacagca gcgcgttggc 540
ccagaagccc tacagcgggc ggaggccaga cgccatcagg ctgggcctgg ggaaccacaa 600
ctactgcaga aacccagatc gagactcaaa gccctggtgc tacgtcttta aggcggggaa 660
gtacagctca gagttctgca gcacccctgc ctgctctgag ggaaacagtg actgctactt 720
tgggaatggg tcagcctacc gtggcacgca cagcctcacc gagtcgggtg cctcctgcct 780
cccgtggaat tccatgatcc tgataggcaa ggtttacaca gcacagaacc ccagtgccca 840
ggcactgggc ctgggcaaac ataattactg ccggaatcct gatggggatg ccaagccctg 900
gtgccacgtg ctgaagaacc gcaggctgac gtgggagtac tgtgatgtgc cctcctgctc 960
cacctgcggc ctgagacagt acagccagcc tcagtttcgc atcaaaggag ggctcttcgc 1020
cgacatcgcc tcccacccct ggcaggctgc catctttgcc aagcacagga ggtcgcccgg 1080
agagcggttc ctgtgcgggg gcatactcat cagctcctgc tggattctct ctgccgccca 1140
ctgcttccag gagaggtttc cgccccacca cctgacggtg atcttgggca gaacataccg 1200
ggtggtccct ggcgaggagg agcagaaatt tgaagtcgaa aaatacattg tccataagga 1260
attcgatgat gacacttacg acaatgacat tgcgctgctg cagctgaaat cggattcgtc 1320
ccgctgtgcc caggagagca gcgtggtccg cactgtgtgc cttcccccgg cggacctgca 1380
gctgccggac tggacggagt gtgagctctc cggctacggc aagcatgagg ccttgtctcc 1440
tttctattcg gagcggctga aggaggctca tgtcagactg tacccatcca gccgctgcac 1500
atcacaacat ttacttaaca gaacagtcac cgacaacatg ctgtgtgctg gagacactcg 1560
gagcggcggg ccccaggcaa acttgcacga cgcctgccag ggcgattcgg gaggccccct 1620
ggtgtgtctg aacgatggcc gcatgacttt ggtgggcatc atcagctggg gcctgggctg 1680
tggacagaag gatgtcccgg gtgtgtacac caaggttacc aactacctag actggattcg 1740
tgacaacatg cgaccgtgac caggaacacc cgactcctca aaagcaaatg agatcccgcc 1800
tcttcttctt cagaagacac tgcaaaggcg cagtgcttct ctacagactt ctccagaccc 1860
accacaccgc agaagcggga cgagacccta caggagaggg aagagtgcat tttcccagat 1920
acttcccatt ttggaagttt tcaggacttg gtctgatttc aggatactct gtcagatggg 1980
aagacatgaa tgcacactag cctctccagg aatgcctcct ccctgggcag aaagtggcca 2040
tgccaccctg ttttcagcta aagcccaacc tcctgacctg tcaccgtgag cagctttgga 2100
aacaggacca caaaaatgaa agcatgtctc aatagtaaaa gataacaaga tctttcagga 2160
aagacggatt gcattagaaa tagacagtat atttatagtc acaagagccc agcagggcct 2220
caaagttggg gcaggctggc tggcccgtca tgttcctcaa aagcaccctt gacgtcaagt 2280
ctccttcccc tttccccact ccctggctct cagaaggtat tccttttgtg tacagtgtgt 2340
aaagtgtaaa tcctttttct ttataaactt tagagtagca tgagagaatt gtatcatttg 2400
aacaactagg cttcagcata tttatagcaa tccatgttag tttttacttt ctgttgccac 2460
aaccctgttt tatactgtac ttaataaatt cagatatatt tttcacagtt tttccaaaaa 2520
aaaaaaaaaa aaaa 2534
<210>197
<211>516
<212>PRT
<213>Homo sapiens
<220>
<223>t-plasminogen activator (tPA)isoform 3
<400>197
Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly
1 5 10 15
Ala Val Phe Val Ser Pro Ser Gln Glu Ile His Ala Arg Phe Arg Arg
20 25 30
Gly Ala Arg Ser Tyr Gln Gly Cys Ser Glu Pro Arg Cys Phe Asn Gly
35 40 45
Gly Thr Cys Gln Gln Ala Leu Tyr Phe Ser Asp Phe Val Cys Gln Cys
50 55 60
Pro Glu Gly Phe Ala Gly Lys Cys Cys Glu Ile Asp Thr Arg Ala Thr
65 70 75 80
Cys Tyr Glu Asp Gln Gly Ile Ser Tyr Arg Gly Thr Trp Ser Thr Ala
85 90 95
Glu Ser Gly Ala Glu Cys Thr Asn Trp Asn Ser Ser Ala Leu Ala Gln
100 105 110
Lys Pro Tyr Ser Gly Arg Arg Pro Asp Ala Ile Arg Leu Gly Leu Gly
115 120 125
Asn His Asn Tyr Cys Arg Asn Pro Asp Arg Asp Ser Lys Pro Trp Cys
130 135 140
Tyr Val Phe Lys Ala Gly Lys Tyr Ser Ser Glu Phe Cys Ser Thr Pro
145 150 155 160
Ala Cys Ser Glu Gly Asn Ser Asp Cys Tyr Phe Gly Asn Gly Ser Ala
165 170 175
Tyr Arg Gly Thr His Ser Leu Thr Glu Ser Gly Ala Ser Cys Leu Pro
180 185 190
Trp Asn Ser Met Ile Leu Ile Gly Lys Val Tyr Thr Ala Gln Asn Pro
195 200 205
Ser Ala Gln Ala Leu Gly Leu Gly Lys His Asn Tyr Cys Arg Asn Pro
210 215 220
Asp Gly Asp Ala Lys Pro Trp Cys His Val Leu Lys Asn Arg Arg Leu
225 230 235 240
Thr Trp Glu Tyr Cys Asp Val Pro Ser Cys Ser Thr Cys Gly Leu Arg
245 250 255
Gln Tyr Ser Gln Pro Gln Phe Arg Ile Lys Gly Gly Leu Phe Ala Asp
260 265 270
Ile Ala Ser His Pro Trp Gln Ala Ala Ile Phe Ala Lys His Arg Arg
275 280 285
Ser Pro Gly Glu Arg Phe Leu Cys Gly Gly Ile Leu Ile Ser Ser Cys
290 295 300
Trp Ile Leu Ser Ala Ala His Cys Phe Gln Glu Arg Phe Pro Pro His
305 310 315 320
His Leu Thr Val Ile Leu Gly Arg Thr Tyr Arg Val Val Pro Gly Glu
325 330 335
Glu Glu Gln Lys Phe Glu Val Glu Lys Tyr Ile Val His Lys Glu Phe
340 345 350
Asp Asp Asp Thr Tyr Asp Asn Asp Ile Ala Leu Leu Gln Leu Lys Ser
355 360 365
Asp Ser Ser Arg Cys Ala Gln Glu Ser Ser Val Val Arg Thr Val Cys
370 375 380
Leu Pro Pro Ala Asp Leu Gln Leu Pro Asp Trp Thr Glu Cys Glu Leu
385 390 395 400
Ser Gly Tyr Gly Lys His Glu Ala Leu Ser Pro Phe Tyr Ser Glu Arg
405 410 415
Leu Lys Glu Ala His Val Arg Leu Tyr Pro Ser Ser Arg Cys Thr Ser
420 425 430
Gln His Leu Leu Asn Arg Thr Val Thr Asp Asn Met Leu Cys Ala Gly
435 440 445
Asp Thr Arg Ser Gly Gly Pro Gln Ala Asn Leu His Asp Ala Cys Gln
450 455 460
Gly Asp Ser Gly Gly Pro Leu Val Cys Leu Asn Asp Gly Arg Met Thr
465 470 475 480
Leu Val Gly Ile Ile Ser Trp Gly Leu Gly Cys Gly Gln Lys Asp Val
485 490 495
Pro Gly Val Tyr Thr Lys Val Thr Asn Tyr Leu Asp Trp Ile Arg Asp
500 505 510
Asn Met Arg Pro
515
<210>198
<211>2732
<212>DNA
<213>Homo sapiens
<220>
<223>Plasmin
<400>198
aacaacatcc tgggattggg acccactttc tgggcactgc tggccagtcc caaaatggaa 60
cataaggaag tggttcttct acttctttta tttctgaaat caggtcaagg agagcctctg 120
gatgactatg tgaataccca gggggcttca ctgttcagtg tcactaagaa gcagctggga 180
gcaggaagta tagaagaatg tgcagcaaaa tgtgaggagg acgaagaatt cacctgcagg 240
gcattccaat atcacagtaa agagcaacaa tgtgtgataa tggctgaaaa caggaagtcc 300
tccataatca ttaggatgag agatgtagtt ttatttgaaa agaaagtgta tctctcagag 360
tgcaagactg ggaatggaaa gaactacaga gggacgatgt ccaaaacaaa aaatggcatc 420
acctgtcaaa aatggagttc cacttctccc cacagaccta gattctcacc tgctacacac 480
ccctcagagg gactggagga gaactactgc aggaatccag acaacgatcc gcaggggccc 540
tggtgctata ctactgatcc agaaaagaga tatgactact gcgacattct tgagtgtgaa 600
gaggaatgta tgcattgcag tggagaaaac tatgacggca aaatttccaa gaccatgtct 660
ggactggaat gccaggcctg ggactctcag agcccacacg ctcatggata cattccttcc 720
aaatttccaa acaagaacct gaagaagaat tactgtcgta accccgatag ggagctgcgg 780
ccttggtgtt tcaccaccga ccccaacaag cgctgggaac tttgcgacat cccccgctgc 840
acaacacctc caccatcttc tggtcccacc taccagtgtc tgaagggaac aggtgaaaac 900
tatcgcggga atgtggctgt taccgtttcc gggcacacct gtcagcactg gagtgcacag 960
acccctcaca cacataacag gacaccagaa aacttcccct gcaaaaattt ggatgaaaac 1020
tactgccgca atcctgacgg aaaaagggcc ccatggtgcc atacaaccaa cagccaagtg 1080
cggtgggagt actgtaagat accgtcctgt gactcctccc cagtatccac ggaacaattg 1140
gctcccacag caccacctga gctaacccct gtggtccagg actgctacca tggtgatgga 1200
cagagctacc gaggcacatc ctccaccacc accacaggaa agaagtgtca gtcttggtca 1260
tctatgacac cacaccggca ccagaagacc ccagaaaact acccaaatgc tggcctgaca 1320
atgaactact gcaggaatcc agatgccgat aaaggcccct ggtgttttac cacagacccc 1380
agcgtcaggt gggagtactg caacctgaaa aaatgctcag gaacagaagc gagtgttgta 1440
gcacctccgc ctgttgtcct gcttccagat gtagagactc cttccgaaga agactgtatg 1500
tttgggaatg ggaaaggata ccgaggcaag agggcgacca ctgttactgg gacgccatgc 1560
caggactggg ctgcccagga gccccataga cacagcattt tcactccaga gacaaatcca 1620
cgggcgggtc tggaaaaaaa ttactgccgt aaccctgatg gtgatgtagg tggtccctgg 1680
tgctacacga caaatccaag aaaactttac gactactgtg atgtccctca gtgtgcggcc 1740
ccttcatttg attgtgggaa gcctcaagtg gagccgaaga aatgtcctgg aagggttgtg 1800
ggggggtgtg tggcccaccc acattcctgg ccctggcaag tcagtcttag aacaaggttt 1860
ggaatgcact tctgtggagg caccttgata tccccagagt gggtgttgac tgctgcccac 1920
tgcttggaga agtccccaag gccttcatcc tacaaggtca tcctgggtgc acaccaagaa 1980
gtgaatctcg aaccgcatgt tcaggaaata gaagtgtcta ggctgttctt ggagcccaca 2040
cgaaaagata ttgccttgct aaagctaagc agtcctgccg tcatcactga caaagtaatc 2100
ccagcttgtc tgccatcccc aaattatgtg gtcgctgacc ggaccgaatg tttcatcact 2160
ggctggggag aaacccaagg tacttttgga gctggccttc tcaaggaagc ccagctccct 2220
gtgattgaga ataaagtgtg caatcgctat gagtttctga atggaagagt ccaatccacc 2280
gaactctgtg ctgggcattt ggccggaggc actgacagtt gccagggtga cagtggaggt 2340
cctctggttt gcttcgagaa ggacaaatac attttacaag gagtcacttc ttggggtctt 2400
ggctgtgcac gccccaataa gcctggtgtc tatgttcgtg tttcaaggtt tgttacttgg 2460
attgagggag tgatgagaaa taattaattg gacgggagac agagtgacgc actgactcac 2520
ctagaggctg ggacgtgggt agggatttag catgctggaa ataactggca gtaatcaaac 2580
gaagacactg tccccagcta ccagctacgc caaacctcgg cattttttgt gttattttct 2640
gactgctgga ttctgtagta aggtgacata gctatgacat ttgttaaaaa taaactctgt 2700
acttaacttt gatttgagta aattttggtt tt 2732
<210>199
<211>810
<212>PRT
<213>Homo sapiens
<220>
<223>Plasmin
<400>199
Met Glu His Lys Glu Val Val Leu Leu Leu Leu Leu Phe Leu Lys Ser
1 5 10 15
Gly Gln Gly Glu Pro Leu Asp Asp Tyr Val Asn Thr Gln Gly Ala Ser
20 25 30
Leu Phe Ser Val Thr Lys Lys Gln Leu Gly Ala Gly Ser Ile Glu Glu
35 40 45
Cys Ala Ala Lys Cys Glu Glu Asp Glu Glu Phe Thr Cys Arg Ala Phe
50 55 60
Gln Tyr His Ser Lys Glu Gln Gln Cys Val Ile Met Ala Glu Asn Arg
65 70 75 80
Lys Ser Ser Ile Ile Ile Arg Met Arg Asp Val Val Leu Phe Glu Lys
85 90 95
Lys ValTyr Leu Ser Glu Cys Lys Thr Gly Asn Gly Lys Asn Tyr Arg
100 105 110
Gly Thr Met Ser Lys Thr Lys Asn Gly Ile Thr Cys Gln Lys Trp Ser
115 120 125
Ser Thr Ser Pro His Arg Pro Arg Phe Ser Pro Ala Thr His Pro Ser
130 135 140
Glu Gly Leu Glu Glu Asn Tyr Cys Arg Asn Pro Asp Asn Asp Pro Gln
145 150 155 160
Gly Pro Trp Cys Tyr Thr Thr Asp Pro Glu Lys Arg Tyr Asp Tyr Cys
165 170 175
Asp Ile Leu Glu Cys Glu Glu Glu Cys Met His Cys Ser Gly Glu Asn
180 185 190
Tyr Asp Gly Lys Ile Ser Lys Thr Met Ser Gly Leu Glu Cys Gln Ala
195 200 205
Trp Asp Ser Gln Ser Pro His Ala His Gly Tyr Ile Pro Ser Lys Phe
210 215 220
Pro Asn Lys Asn Leu Lys Lys Asn Tyr Cys Arg Asn Pro Asp Arg Glu
225 230 235 240
Leu Arg Pro Trp Cys Phe Thr Thr Asp Pro Asn Lys Arg Trp Glu Leu
245 250 255
Cys Asp Ile Pro Arg Cys Thr Thr Pro Pro Pro Ser Ser Gly Pro Thr
260 265 270
Tyr Gln Cys Leu Lys Gly Thr Gly Glu Asn Tyr Arg Gly Asn Val Ala
275 280 285
Val Thr Val Ser Gly His Thr Cys Gln His Trp Ser Ala Gln Thr Pro
290 295 300
His Thr His Asn Arg Thr Pro Glu Asn Phe Pro Cys Lys Asn Leu Asp
305 310 315 320
Glu Asn Tyr Cys Arg Asn Pro Asp Gly Lys Arg Ala Pro Trp Cys His
325 330 335
Thr Thr Asn Ser Gln Val Arg Trp Glu Tyr Cys Lys Ile Pro Ser Cys
340 345 350
Asp Ser Ser Pro Val Ser Thr Glu Gln Leu Ala Pro Thr Ala Pro Pro
355 360 365
Glu Leu Thr Pro Val Val Gln Asp Cys Tyr His Gly Asp Gly Gln Ser
370 375 380
Tyr Arg Gly Thr Ser Ser Thr Thr Thr Thr Gly Lys Lys Cys Gln Ser
385 390 395 400
Trp Ser Ser Met Thr Pro His Arg His Gln Lys Thr Pro Glu Asn Tyr
405 410 415
Pro Asn Ala Gly Leu Thr Met Asn Tyr Cys Arg Asn Pro Asp Ala Asp
420 425 430
Lys Gly Pro Trp Cys Phe Thr Thr Asp Pro Ser Val Arg Trp Glu Tyr
435 440 445
Cys Asn Leu Lys Lys Cys Ser Gly Thr Glu Ala Ser Val Val Ala Pro
450 455 460
Pro Pro Val Val Leu Leu Pro Asp Val Glu Thr Pro Ser Glu Glu Asp
465 470 475 480
Cys Met Phe Gly Asn Gly Lys Gly Tyr Arg Gly Lys Arg Ala Thr Thr
485 490 495
Val Thr Gly Thr Pro Cys Gln Asp Trp Ala Ala Gln Glu Pro His Arg
500 505 510
His Ser Ile Phe Thr Pro Glu Thr Asn Pro Arg Ala Gly Leu Glu Lys
515 520 525
Asn Tyr Cys Arg Asn Pro Asp Gly Asp Val Gly Gly Pro Trp Cys Tyr
530 535 540
Thr Thr Asn Pro Arg Lys Leu Tyr Asp Tyr Cys Asp Val Pro Gln Cys
545 550 555 560
Ala Ala Pro Ser Phe Asp Cys Gly Lys Pro Gln Val Glu Pro Lys Lys
565 570 575
Cys Pro Gly Arg Val Val Gly Gly Cys Val Ala His Pro His Ser Trp
580 585 590
Pro Trp Gln Val Ser Leu Arg Thr Arg Phe Gly Met His Phe Cys Gly
595 600 605
Gly Thr Leu Ile Ser Pro Glu Trp Val Leu Thr Ala Ala His Cys Leu
610 615 620
Glu Lys Ser Pro Arg Pro Ser Ser Tyr Lys Val Ile Leu Gly Ala His
625 630 635 640
Gln Glu Val Asn Leu Glu Pro His Val Gln Glu Ile Glu Val Ser Arg
645 650 655
Leu Phe Leu Glu Pro Thr Arg Lys Asp Ile Ala Leu Leu Lys Leu Ser
660 665 670
Ser Pro Ala Val Ile Thr Asp Lys Val Ile Pro Ala Cys Leu Pro Ser
675 680 685
Pro Asn Tyr Val Val Ala Asp Arg Thr Glu Cys Phe Ile Thr Gly Trp
690 695 700
Gly Glu Thr Gln Gly Thr Phe Gly Ala Gly Leu Leu Lys Glu Ala Gln
705 710 715 720
Leu Pro Val Ile Glu Asn Lys Val Cys Asn Arg Tyr Glu Phe Leu Asn
725 730 735
Gly Arg Val Gln Ser Thr Glu Leu Cys Ala Gly His Leu Ala Gly Gly
740 745 750
Thr Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Phe Glu
755 760 765
Lys Asp Lys Tyr Ile Leu Gln Gly Val Thr Ser Trp Gly Leu Gly Cys
770 775 780
Ala Arg Pro Asn Lys Pro Gly Val Tyr Val Arg Val Ser Arg Phe Val
785 790 795 800
Thr Trp Ile Glu Gly Val Met Arg Asn Asn
805 810
<210>200
<211>1527
<212>DNA
<213>Homo sapiens
<220>
<223>Neurosin transcript variant A
<400>200
ggcggacaaa gcccgattgt tcctgggccc tttccccatc gcgcctgggc ctgctcccca 60
gcccggggca ggggcggggg ccagtgtggt gacacacgct gtagctgtct ccccggctgg 120
ctggctcgct ctctcctggg gacacagagg tcggcaggca gcacacagag ggacctacgg 180
gcagctgttc cttcccccga ctcaagaatc cccggaggcc cggaggcctg cagcaggagc 240
ggccatgaag aagctgatgg tggtgctgag tctgattgct gcagcctggg cagaggagca 300
gaataagttg gtgcatggcg gaccctgcga caagacatct cacccctacc aagctgccct 360
ctacacctcg ggccacttgc tctgtggtgg ggtccttatc catccactgt gggtcctcac 420
agctgcccac tgcaaaaaac cgaatcttca ggtcttcctg gggaagcata accttcggca 480
aagggagagt tcccaggagc agagttctgt tgtccgggct gtgatccacc ctgactatga 540
tgccgccagc catgaccagg acatcatgct gttgcgcctg gcacgcccag ccaaactctc 600
tgaactcatc cagccccttc ccctggagag ggactgctca gccaacacca ccagctgcca 660
catcctgggc tggggcaaga cagcagatgg tgatttccct gacaccatcc agtgtgcata 720
catccacctg gtgtcccgtg aggagtgtga gcatgcctac cctggccaga tcacccagaa 780
catgttgtgt gctggggatg agaagtacgg gaaggattcc tgccagggtg attctggggg 840
tccgctggta tgtggagacc acctccgagg ccttgtgtca tggggtaaca tcccctgtgg 900
atcaaaggag aagccaggag tctacaccaa cgtctgcaga tacacgaact ggatccaaaa 960
aaccattcag gccaagtgac cctgacatgt gacatctacc tcccgaccta ccaccccact 1020
ggctggttcc agaacgtctc tcacctagac cttgcctccc ctcctctcct gcccagctct 1080
gaccctgatg cttaataaac gcagcgacgt gagggtcctg attctccctg gttttacccc 1140
agctccatcc ttgcatcact ggggaggacg tgatgagtga ggacttgggt cctcggtctt 1200
acccccacca ctaagagaat acaggaaaat cccttctagg catctcctct ccccaaccct 1260
tccacacgtt tgatttcttc ctgcagaggc ccagccacgt gtctggaatc ccagctccgc 1320
tgcttactgt cggtgtcccc ttgggatgta cctttcttca ctgcagattt ctcacctgta 1380
agatgaagat aaggatgata cagtctccat aaggcagtgg ctgttggaaa gatttaaggt 1440
ttcacaccta tgacatacat ggaatagcac ctgggccacc atgcactcaa taaagaatga 1500
attttattat gaaaaaaaaa aaaaaaa 1527
<210>201
<211>244
<212>PRT
<213>Homo sapiens
<220>
<223>Neurosin isoform A
<400>201
Met Lys Lys Leu Met Val Val Leu Ser Leu Ile Ala Ala Ala Trp Ala
1 5 10 15
Glu Glu Gln Asn Lys Leu Val His Gly Gly Pro Cys Asp Lys Thr Ser
20 25 30
His Pro Tyr Gln Ala Ala Leu Tyr Thr Ser Gly His Leu Leu Cys Gly
35 40 45
Gly Val Leu Ile His Pro Leu Trp Val Leu Thr Ala Ala His Cys Lys
50 55 60
Lys Pro Asn Leu Gln Val Phe Leu Gly Lys His Asn Leu Arg Gln Arg
65 70 75 80
Glu Ser Ser Gln Glu Gln Ser Ser Val Val Arg Ala Val Ile His Pro
85 90 95
Asp Tyr Asp Ala Ala Ser His Asp Gln Asp Ile Met Leu Leu Arg Leu
100 105 110
Ala Arg Pro Ala Lys Leu Ser Glu Leu Ile Gln Pro Leu Pro Leu Glu
115 120 125
Arg Asp Cys Ser Ala Asn Thr Thr Ser Cys His Ile Leu Gly Trp Gly
130 135 140
Lys Thr Ala Asp Gly Asp Phe Pro Asp Thr Ile Gln Cys Ala Tyr Ile
145 150 155 160
His Leu Val Ser Arg Glu Glu Cys Glu His Ala Tyr Pro Gly Gln Ile
165 170 175
Thr Gln Asn Met Leu Cys Ala Gly Asp Glu Lys Tyr Gly Lys Asp Ser
180 185 190
Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Gly Asp His Leu Arg
195 200 205
Gly Leu ValSer Trp Gly Asn Ile Pro Cys Gly Ser Lys Glu Lys Pro
210 215 220
Gly Val Tyr Thr Asn Val Cys Arg Tyr Thr Asn Trp Ile Gln Lys Thr
225 230 235 240
Ile Gln Ala Lys
<210>202
<211>1527
<212>DNA
<213>Homo sapiens
<220>
<223>Neurosin transcript variant B
<400>202
aggacgttcc agaagcatct ggggacagaa ccagcctctt ccagggaggc ctgggagctg 60
ggggtgtgtg tctggcagtc cctgcagccc tgggctctgc ggcccctgcg tcctccgctt 120
ggctctgcca ctgcatctga gtgtcttctc tcctcacggc tccccgcatt tctaactctt 180
tctgcctcct cgtctcaaag ctgttccttc ccccgactca agaatccccg gaggcccgga 240
ggcctgcagc aggagcggcc atgaagaagc tgatggtggt gctgagtctg attgctgcag 300
cctgggcaga ggagcagaat aagttggtgc atggcggacc ctgcgacaag acatctcacc 360
cctaccaagc tgccctctac acctcgggcc acttgctctg tggtggggtc cttatccatc 420
cactgtgggt cctcacagct gcccactgca aaaaaccgaa tcttcaggtc ttcctgggga 480
agcataacct tcggcaaagg gagagttccc aggagcagag ttctgttgtc cgggctgtga 540
tccaccctga ctatgatgcc gccagccatg accaggacat catgctgttg cgcctggcac 600
gcccagccaa actctctgaa ctcatccagc cccttcccct ggagagggac tgctcagcca 660
acaccaccag ctgccacatc ctgggctggg gcaagacagc agatggtgat ttccctgaca 720
ccatccagtg tgcatacatc cacctggtgt cccgtgagga gtgtgagcat gcctaccctg 780
gccagatcac ccagaacatg ttgtgtgctg gggatgagaa gtacgggaag gattcctgcc 840
agggtgattc tgggggtccg ctggtatgtg gagaccacct ccgaggcctt gtgtcatggg 900
gtaacatccc ctgtggatca aaggagaagc caggagtcta caccaacgtc tgcagataca 960
cgaactggat ccaaaaaacc attcaggcca agtgaccctg acatgtgaca tctacctccc 1020
gacctaccac cccactggct ggttccagaa cgtctctcac ctagaccttg cctcccctcc 1080
tctcctgccc agctctgacc ctgatgctta ataaacgcag cgacgtgagg gtcctgattc 1140
tccctggttt taccccagct ccatccttgc atcactgggg aggacgtgat gagtgaggac 1200
ttgggtcctc ggtcttaccc ccaccactaa gagaatacag gaaaatccct tctaggcatc 1260
tcctctcccc aacccttcca cacgtttgat ttcttcctgc agaggcccag ccacgtgtct 1320
ggaatcccag ctccgctgct tactgtcggt gtccccttgg gatgtacctt tcttcactgc 1380
agatttctca cctgtaagat gaagataagg atgatacagt ctccataagg cagtggctgt 1440
tggaaagatt taaggtttca cacctatgac atacatggaa tagcacctgg gccaccatgc 1500
actcaataaa gaatgaattt tattatg 1527
<210>203
<211>244
<212>PRT
<213>Homo sapiens
<220>
<223>Neurosin isoform A preprotein
<400>203
Met Lys Lys Leu Met Val Val Leu Ser Leu Ile Ala Ala Ala Trp Ala
1 5 10 15
Glu Glu Gln Asn Lys Leu Val His Gly Gly Pro Cys Asp Lys Thr Ser
20 25 30
His Pro Tyr Gln Ala Ala Leu Tyr Thr Ser Gly His Leu Leu Cys Gly
35 40 45
Gly Val Leu Ile His Pro Leu Trp Val Leu Thr Ala Ala His Cys Lys
50 55 60
Lys Pro Asn Leu Gln Val Phe Leu Gly Lys His Asn Leu Arg Gln Arg
65 70 75 80
Glu Ser Ser Gln Glu Gln Ser Ser ValVal Arg Ala Val Ile His Pro
85 90 95
Asp Tyr Asp Ala Ala Ser His Asp Gln Asp Ile Met Leu Leu Arg Leu
100 105 110
Ala Arg Pro Ala Lys Leu Ser Glu Leu Ile Gln Pro Leu Pro Leu Glu
115 120 125
Arg Asp Cys Ser Ala Asn Thr Thr Ser Cys His Ile Leu Gly Trp Gly
130 135 140
Lys Thr Ala Asp Gly Asp Phe Pro Asp Thr Ile Gln Cys Ala Tyr Ile
145 150 155 160
His Leu Val Ser Arg Glu Glu Cys Glu His Ala Tyr Pro Gly Gln Ile
165 170 175
Thr Gln Asn Met Leu Cys Ala Gly Asp Glu Lys Tyr Gly Lys Asp Ser
180 185 190
Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Gly Asp His Leu Arg
195 200 205
Gly Leu Val Ser Trp Gly Asn Ile Pro Cys Gly Ser Lys Glu Lys Pro
210 215 220
Gly Val Tyr Thr Asn Val Cys Arg Tyr Thr Asn Trp Ile Gln Lys Thr
225 230 235 240
Ile Gln Ala Lys
<210>204
<211>1495
<212>DNA
<213>Homo sapiens
<220>
<223>Neurosin transcript variant C
<400>204
aggacgttcc agaagcatct ggggacagaa ccagcctctt ccagggaggc ctgggagctg 60
ggggtgtgtg tctggcagtc cctgcagccc tgggctctgc ggcccctgcg tcctccgctt 120
ggctctgcca ctgcatctga gtgtcttctc tcctcacggc tccccgcatt tctaactctt 180
tctgcctcct cgtctcaaag ctgttccttc ccccgactca agaatccccg gaggcccgga 240
ggcctgcagc agcctgggca gaggagcaga ataagttggt gcatggcgga ccctgcgaca 300
agacatctca cccctaccaa gctgccctct acacctcggg ccacttgctc tgtggtgggg 360
tccttatcca tccactgtgg gtcctcacag ctgcccactg caaaaaaccg aatcttcagg 420
tcttcctggg gaagcataac cttcggcaaa gggagagttc ccaggagcag agttctgttg 480
tccgggctgt gatccaccct gactatgatg ccgccagcca tgaccaggac atcatgctgt 540
tgcgcctggc acgcccagcc aaactctctg aactcatcca gccccttccc ctggagaggg 600
actgctcagc caacaccacc agctgccaca tcctgggctg gggcaagaca gcagatggtg 660
atttccctga caccatccag tgtgcataca tccacctggt gtcccgtgag gagtgtgagc 720
atgcctaccc tggccagatc acccagaaca tgttgtgtgc tggggatgag aagtacggga 780
aggattcctg ccagggtgat tctgggggtc cgctggtatg tggagaccac ctccgaggcc 840
ttgtgtcatg gggtaacatc ccctgtggat caaaggagaa gccaggagtc tacaccaacg 900
tctgcagata cacgaactgg atccaaaaaa ccattcaggc caagtgaccc tgacatgtga 960
catctacctc ccgacctacc accccactgg ctggttccag aacgtctctc acctagacct 1020
tgcctcccct cctctcctgc ccagctctga ccctgatgct taataaacgc agcgacgtga 1080
gggtcctgat tctccctggt tttaccccag ctccatcctt gcatcactgg ggaggacgtg 1140
atgagtgagg acttgggtcc tcggtcttac ccccaccact aagagaatac aggaaaatcc 1200
cttctaggca tctcctctcc ccaacccttc cacacgtttg atttcttcct gcagaggccc 1260
agccacgtgt ctggaatccc agctccgctg cttactgtcg gtgtcccctt gggatgtacc 1320
tttcttcact gcagatttct cacctgtaag atgaagataa ggatgataca gtctccataa 1380
ggcagtggct gttggaaaga tttaaggttt cacacctatg acatacatgg aatagcacct 1440
gggccaccat gcactcaata aagaatgaat tttattatga aaaaaaaaaa aaaaa 1495
<210>205
<211>137
<212>PRT
<213>Homo sapiens
<220>
<223>Neurosin isoform B
<400>205
Met Leu Leu Arg Leu Ala Arg Pro Ala Lys Leu Ser Glu Leu Ile Gln
1 5 10 15
Pro Leu Pro Leu Glu Arg Asp Cys Ser Ala Asn Thr Thr Ser Cys His
20 25 30
Ile Leu Gly Trp Gly Lys Thr Ala Asp Gly Asp Phe Pro Asp Thr Ile
35 40 45
Gln Cys Ala Tyr Ile His Leu Val Ser Arg Glu Glu Cys Glu His Ala
50 55 60
Tyr Pro Gly Gln Ile Thr Gln Asn Met Leu Cys Ala Gly Asp Glu Lys
65 70 75 80
Tyr Gly Lys Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys
85 90 95
Gly Asp His Leu Arg Gly Leu Val Ser Trp Gly Asn Ile Pro Cys Gly
100 105 110
Ser Lys Glu Lys Pro Gly Val Tyr Thr Asn Val Cys Arg Tyr Thr Asn
115 120 125
Trp Ile Gln Lys Thr Ile Gln Ala Lys
130 135
<210>206
<211>1386
<212>DNA
<213>Homo sapiens
<220>
<223>Neurosin transcript variant D
<400>206
aggacgttcc agaagcatct ggggacagaa ccagcctctt ccagggaggc ctgggagctg 60
ggggtgtgtg tctggcagtc cctgcagccc tgggctctgc ggcccctgcg tcctccgctt 120
ggctctgcca ctgcatctga gtgtcttctc tcctcacggc tccccgcatt tctaactctt 180
tctgcctcct cgtctcaaag ctgttccttc ccccgactca agaatccccg gaggcccgga 240
ggcctgcagc aggagcggcc atgaagaagc tgatggtggt gctgagtctg attgctgcag 300
gaatcttcag gtcttcctgg ggaagcataa ccttcggcaa agggagagtt cccaggagca 360
gagttctgtt gtccgggctg tgatccaccc tgactatgat gccgccagcc atgaccagga 420
catcatgctg ttgcgcctgg cacgcccagc caaactctct gaactcatcc agccccttcc 480
cctggagagg gactgctcag ccaacaccac cagctgccac atcctgggct ggggcaagac 540
agcagatggt gatttccctg acaccatcca gtgtgcatac atccacctgg tgtcccgtga 600
ggagtgtgag catgcctacc ctggccagat cacccagaac atgttgtgtg ctggggatga 660
gaagtacggg aaggattcct gccagggtga ttctgggggt ccgctggtat gtggagacca 720
cctccgaggc cttgtgtcat ggggtaacat cccctgtgga tcaaaggaga agccaggagt 780
ctacaccaac gtctgcagat acacgaactg gatccaaaaa accattcagg ccaagtgacc 840
ctgacatgtg acatctacct cccgacctac caccccactg gctggttcca gaacgtctct 900
cacctagacc ttgcctcccc tcctctcctg cccagctctg accctgatgc ttaataaacg 960
cagcgacgtg agggtcctga ttctccctgg ttttacccca gctccatcct tgcatcactg 1020
gggaggacgt gatgagtgag gacttgggtc ctcggtctta cccccaccac taagagaata 1080
caggaaaatc ccttctaggc atctcctctc cccaaccctt ccacacgttt gatttcttcc 1140
tgcagaggcc cagccacgtg tctggaatcc cagctccgct gcttactgtc ggtgtcccct 1200
tgggatgtac ctttcttcac tgcagatttc tcacctgtaa gatgaagata aggatgatac 1260
agtctccata aggcagtggc tgttggaaag atttaaggtt tcacacctat gacatacatg 1320
gaatagcacc tgggccacca tgcactcaat aaagaatgaa ttttattatg aaaaaaaaaa 1380
aaaaaa 1386
<210>207
<211>40
<212>PRT
<213>Homo sapiens
<220>
<223>Neurosin isoform C
<400>207
Met Lys Lys Leu Met Val Val Leu Ser Leu Ile Ala Ala Gly Ile Phe
1 5 10 15
Arg Ser Ser Trp Gly Ser Ile Thr Phe Gly Lys Gly Arg Val Pro Arg
20 25 30
Ser Arg Val Leu Leu Ser Gly Leu
35 40
<210>208
<211>3351
<212>DNA
<213>Homo sapiens
<220>
<223>Neurotrypsin
<400>208
gcccgagaag ctggggagca tggaccagac cccgcagcgc tggcaccatg acgctcgccc 60
gcttcgtgct agccctgatg ttaggggcgc tccccgaagt ggtcggcttt gattctgtcc 120
tcaatgattc cctccaccac agccaccgcc attcgccccc tgcgggtccg cactacccct 180
attaccttcc cacccagcag cggcccccga cgacgcgtcc gccgccgcct ctcccgcgct 240
tcccgcgccc cccgcgggcg ctccctgccc agcgcccgca cgccctccag gccgggcaca 300
cgccccggcc gcacccctgg ggctgccccg ccggcgagcc atgggtcagc gtgacggact 360
tcggcgcccc gtgtctgcgg tgggcggagg tgccaccctt cctggagcgg tcgcccccag 420
cgagctgggc tcagctgcga ggacagcgcc acaacttttg tcggagcccc gacggcgcgg 480
gcagaccctg gtgtttctac ggagacgccc gtggcaaggt ggactggggc tactgcgact 540
gcagacacgg atcagtacga cttcgtggcg gcaaaaatga gtttgaaggc acagtggaag 600
tatatgcaag tggagtttgg ggcactgtct gtagcagcca ctgggatgat tctgatgcat 660
cagtcatttg tcaccagctg cagctgggag gaaaaggaat agcaaaacaa accccgtttt 720
ctggactggg ccttattccc atttattgga gcaatgtccg ttgccgagga gatgaagaaa 780
atatactgct ttgtgaaaaa gacatctggc agggtggggt gtgtcctcag aagatggcag 840
ctgctgtcac gtgtagcttt tcccatggcc caacgttccc catcattcgc cttgctggag 900
gcagcagtgt gcatgaaggc cgggtggagc tctaccatgc tggccagtgg ggaaccgttt 960
gtgatgacca atgggatgat gccgatgcag aagtgatctg caggcagctg ggcctcagtg 1020
gcattgccaa agcatggcat caggcatatt ttggggaagg gtctggccca gttatgttgg 1080
atgaagtacg ctgcactggg aatgagcttt caattgagca gtgtccaaag agctcctggg 1140
gagagcataa ctgtggccat aaagaagatg ctggagtgtc ctgtacccct ctaacagatg 1200
gggtcatcag acttgcaggt gggaaaggca gccatgaggg tcgcttggag gtatattaca 1260
gaggccagtg gggaactgtc tgtgatgatg gctggactga gctgaataca tacgtggttt 1320
gtcgacagtt gggatttaaa tatggtaaac aagcatctgc caaccatttt gaagaaagca 1380
cagggcccat atggttggat gacgtcagct gctcaggaaa ggaaaccaga tttcttcagt 1440
gttccaggcg acagtgggga aggcatgact gcagccaccg cgaagatgtt agcattgcct 1500
gctaccctgg cggcgaggga cacaggctct ctctgggttt tcctgtcaga ctgatggatg 1560
gagaaaataa gaaagaagga cgagtggagg tttttatcaa tggccagtgg ggaacaatct 1620
gtgatgatgg atggactgat aaggatgcag ctgtgatctg tcgtcagctt ggctacaagg 1680
gtcctgccag agcaagaacc atggcttact ttggagaagg aaaaggaccc atccatgtgg 1740
ataatgtgaa gtgcacagga aatgagaggt ccttggctga ctgtatcaag caagatattg 1800
gaagacacaa ctgccgccac agtgaagatg caggagttat ttgtgattat tttggcaaga 1860
aggcctcagg taacagtaat aaagagtccc tctcatctgt ttgtggcttg agattactgc 1920
accgtcggca gaagcggatc attggtggga aaaattcttt aaggggtggt tggccttggc 1980
aggtttccct ccggctgaag tcatcccatg gagatggcag gctcctctgc ggggctacgc 2040
tcctgagtag ctgctgggtc ctcacagcag cacactgttt caagaggtat ggcaacagca 2100
ctaggagcta tgctgttagg gttggagatt atcatactct ggtaccagag gagtttgagg 2160
aagaaattgg agttcaacag attgtgattc atcgggagta tcgacccgac cgcagtgatt 2220
atgacatagc cctggttaga ttacaaggac cagaagagca atgtgccaga ttcagcagcc 2280
atgttttgcc agcctgttta ccactctgga gagagaggcc acagaaaaca gcatccaact 2340
gttacataac aggatggggt gacacaggac gagcctattc aagaacacta caacaagcag 2400
ccattccctt acttcctaaa aggttttgtg aagaacgtta taagggtcgg tttacaggga 2460
gaatgctttg tgctggaaac ctccatgaac acaaacgcgt ggacagctgc cagggagaca 2520
gcggaggacc actcatgtgt gaacggcccg gagagagctg ggtggtgtat ggggtgacct 2580
cctgggggta tggctgtgga gtcaaggatt ctcctggtgt ttataccaaa gtctcagcct 2640
ttgtaccttg gataaaaagt gtcaccaaac tgtaattctt catggaaact tcaaagcagc 2700
atttaaacaa atggaaaact ttgaaccccc actattagca ctcagcagag atgacaacaa 2760
atggcaagat ctgtttttgc tttgtgttgt ggtaaaaaat tgtgtacccc ctgctgcttt 2820
tgagaaattt gtgaacattt tcagaggcct cagtgtagtg gaagtgataa tccttaaatg 2880
aacattttct accctaattt cactggagtg acttattcta agcctcatct atcccctacc 2940
tatttctcaa aatcattcta tgctgatttt acaaaagatc atttttacat ttgaactgag 3000
aacccctttt aattgaatca gtggtgtctg aaatcatatt aaatacccac atttgacata 3060
aatgcggtac cctttactac actcatgagt ggcatattta tgcttaggtc ttttcaaaag 3120
acttgacaag aaatcttcat attctctgta gcctttgtca agtgaggaaa tcagtggtta 3180
aagaattcca ctataaactt ttaggcctga ataggagtag taaagcctca aggacatctg 3240
cctgtcacaa tatattctca aagtgatctg atatttggaa acaagtatcc ttgttgagta 3300
ccaagtgcta cagaaaccat aagataaaaa tactttctac ctacagcgtg c 3351
<210>209
<211>875
<212>PRT
<213>Homo sapiens
<220>
<223>Neurotrypsin
<400>209
Met Thr Leu Ala Arg Phe Val Leu Ala Leu Met Leu Gly Ala Leu Pro
1 5 10 15
Glu Val Val Gly Phe Asp Ser Val Leu Asn Asp Ser Leu His His Ser
20 25 30
His Arg His Ser Pro Pro Ala Gly Pro His Tyr Pro Tyr Tyr Leu Pro
35 40 45
Thr Gln Gln Arg Pro Pro Thr Thr Arg Pro Pro Pro Pro Leu Pro Arg
50 55 60
Phe Pro Arg Pro Pro Arg Ala Leu Pro Ala Gln Arg Pro His Ala Leu
65 70 75 80
Gln Ala Gly His Thr Pro Arg Pro His Pro Trp Gly Cys Pro Ala Gly
85 90 95
Glu Pro Trp Val Ser Val Thr Asp Phe Gly Ala Pro Cys Leu Arg Trp
100 105 110
Ala Glu Val Pro Pro Phe Leu Glu Arg Ser Pro Pro Ala Ser Trp Ala
115 120 125
Gln Leu Arg Gly Gln Arg His Asn Phe Cys Arg Ser Pro Asp Gly Ala
130 135 140
Gly Arg Pro Trp Cys Phe Tyr Gly Asp Ala Arg Gly Lys Val Asp Trp
145 150 155 160
Gly Tyr Cys Asp Cys Arg His Gly Ser Val Arg Leu Arg Gly Gly Lys
165 170 175
Asn Glu Phe Glu Gly Thr Val Glu Val Tyr Ala Ser Gly Val Trp Gly
180 185 190
Thr Val Cys Ser Ser His Trp Asp Asp Ser Asp Ala Ser Val Ile Cys
195 200 205
His Gln Leu Gln Leu Gly Gly Lys Gly Ile Ala Lys Gln Thr Pro Phe
210 215 220
Ser Gly Leu Gly Leu Ile Pro Ile Tyr Trp Ser Asn Val Arg Cys Arg
225 230 235 240
Gly Asp Glu Glu Asn Ile Leu Leu Cys Glu Lys Asp Ile Trp Gln Gly
245 250 255
Gly Val Cys Pro Gln Lys Met Ala Ala Ala Val Thr Cys Ser Phe Ser
260 265 270
His Gly Pro Thr Phe Pro Ile Ile Arg Leu Ala Gly Gly Ser Ser Val
275 280 285
His Glu Gly Arg Val Glu Leu Tyr His Ala Gly Gln Trp Gly Thr Val
290 295 300
Cys Asp Asp Gln Trp Asp Asp Ala Asp Ala Glu Val Ile Cys Arg Gln
305 310 315 320
Leu Gly Leu Ser Gly Ile Ala Lys Ala Trp His Gln Ala Tyr Phe Gly
325 330 335
Glu Gly Ser Gly Pro Val Met Leu Asp Glu Val Arg Cys Thr Gly Asn
340 345 350
Glu Leu Ser Ile Glu Gln Cys Pro Lys Ser Ser Trp Gly Glu His Asn
355 360 365
Cys Gly His Lys Glu Asp Ala Gly Val Ser Cys Thr Pro Leu Thr Asp
370 375 380
Gly Val Ile Arg Leu Ala Gly Gly Lys Gly Ser His Glu Gly Arg Leu
385 390 395 400
Glu Val Tyr Tyr Arg Gly Gln Trp Gly Thr Val Cys Asp Asp Gly Trp
405 410 415
Thr Glu Leu Asn Thr Tyr Val Val Cys Arg Gln Leu Gly Phe Lys Tyr
420 425 430
Gly Lys Gln Ala Ser Ala Asn His Phe Glu Glu Ser Thr Gly Pro Ile
435 440 445
Trp Leu Asp Asp Val Ser Cys Ser Gly Lys Glu Thr Arg Phe Leu Gln
450 455 460
Cys Ser Arg Arg Gln Trp Gly Arg His Asp Cys Ser His Arg Glu Asp
465 470 475 480
Val Ser Ile Ala Cys Tyr Pro Gly Gly Glu Gly His Arg Leu Ser Leu
485 490 495
Gly Phe Pro Val Arg Leu Met Asp Gly Glu Asn Lys Lys Glu Gly Arg
500 505 510
Val Glu Val Phe Ile Asn Gly Gln Trp Gly Thr Ile Cys Asp Asp Gly
515 520 525
Trp Thr Asp Lys Asp Ala Ala Val Ile Cys Arg Gln Leu Gly Tyr Lys
530 535 540
Gly Pro Ala Arg Ala Arg Thr Met Ala Tyr Phe Gly Glu Gly Lys Gly
545 550 555 560
Pro Ile His Val Asp Asn Val Lys Cys Thr Gly Asn Glu Arg Ser Leu
565 570 575
Ala Asp Cys Ile Lys Gln Asp Ile Gly Arg His Asn Cys Arg His Ser
580 585 590
Glu Asp Ala Gly Val Ile Cys Asp Tyr Phe Gly Lys Lys Ala Ser Gly
595 600 605
Asn Ser Asn Lys Glu Ser Leu Ser Ser Val Cys Gly Leu Arg Leu Leu
610 615 620
His Arg Arg Gln Lys Arg Ile Ile Gly Gly Lys Asn Ser Leu Arg Gly
625 630 635 640
Gly Trp Pro Trp Gln Val Ser Leu Arg Leu Lys Ser Ser His Gly Asp
645 650 655
Gly Arg Leu Leu Cys Gly Ala Thr Leu Leu Ser Ser Cys Trp Val Leu
660 665 670
Thr Ala Ala His Cys Phe Lys Arg Tyr Gly Asn Ser Thr Arg Ser Tyr
675 680 685
Ala Val Arg Val Gly Asp Tyr His Thr Leu Val Pro Glu Glu Phe Glu
690 695 700
Glu Glu Ile Gly Val Gln Gln Ile Val Ile His Arg Glu Tyr Arg Pro
705 710 715 720
Asp Arg Ser Asp Tyr Asp Ile Ala Leu Val Arg Leu Gln Gly Pro Glu
725 730 735
Glu Gln Cys Ala Arg Phe Ser Ser His Val Leu Pro Ala Cys Leu Pro
740 745 750
Leu Trp Arg Glu Arg Pro Gln Lys Thr Ala Ser Asn Cys Tyr Ile Thr
755 760 765
Gly Trp Gly Asp Thr Gly Arg Ala Tyr Ser Arg Thr Leu Gln Gln Ala
770 775 780
Ala Ile Pro Leu Leu Pro Lys Arg Phe Cys Glu Glu Arg Tyr Lys Gly
785 790 795 800
Arg Phe Thr Gly Arg Met Leu Cys Ala Gly Asn Leu His Glu His Lys
805 810 815
Arg Val Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Met Cys Glu
820 825 830
Arg Pro Gly Glu Ser Trp Val Val Tyr Gly Val Thr Ser Trp Gly Tyr
835 840 845
Gly Cys Gly Val Lys Asp Ser Pro Gly Val Tyr Thr Lys Val Ser Ala
850 855 860
Phe Val Pro Trp Ile Lys Ser Val Thr Lys Leu
865 870 875
<210>210
<211>1165
<212>DNA
<213>Homo sapiens
<220>
<223>tryptase beta 2(I)
<400>210
agaaggaaca gggagcggcc aggatgctga atctgctgct gctggcgctg cccgtcctgg 60
cgagccgcgc ctacgcggcc cctgccccag gccaggccct gcagcgagtg ggcatcgttg 120
ggggtcagga ggcccccagg agcaagtggc cctggcaggt gagcctgaga gtccgcgacc 180
gatactggat gcacttctgc gggggctccc tcatccaccc ccagtgggtg ctgaccgcag 240
cgcactgcgt gggaccggac gtcaaggatc tggccgccct cagggtgcaa ctgcgggagc 300
agcacctcta ctaccaggac cagctgctgc cggtcagcag gatcatcgtg cacccacagt 360
tctacaccgc ccagatcgga gcggacatcg ccctgctgga gctggaggag ccggtgaacg 420
tctccagcca cgtccacacg gtcaccctgc cccctgcctc agagaccttc cccccgggga 480
tgccgtgctg ggtcactggc tggggcgatg tggacaatga tgagcgcctc ccaccgccat 540
ttcctctgaa gcaggtgaag gtccccataa tggaaaacca catttgtgac gcaaaatacc 600
accttggcgc ctacacggga gacgacgtcc gcatcgtccg tgacgacatg ctgtgtgccg 660
ggaacacccg gagggactca tgccagggcg actccggagg gcccctggtg tgcaaggtga 720
atggcacctg gctgcaggcg ggcgtggtca gctggggcga gggctgtgcc cagcccaacc 780
ggcctggcat ctacacccgt gtcacctact acttggactg gatccaccac tatgtcccca 840
aaaagccgtg agtcaggcct ggggtgtcca cctgggtcac tggagagcca gcccctcctg 900
tccaaaacac cactgcttcc tacccaggtg gcgactgccc cccacacctt ccctgccccg 960
tcctgagtgc cccttcctgt cctaagcccc ctgctctctt ctgagcccct tcccctgtcc 1020
tgaggaccct tccccatcct gagccccctt ccctgtccta agcctgacgc ctgcaccggg 1080
ccctccggcc ctcccctgcc caggcagctg gtggtgggcg ctaatcctcc tgagtgctgg 1140
acctcattaa agtgcatgga aatca 1165
<210>211
<211>275
<212>PRT
<213>Homo sapiens
<220>
<223>tryptase beta 2(I)
<400>211
Met Leu Asn Leu Leu Leu Leu Ala Leu Pro Val Leu Ala Ser Arg Ala
1 5 10 15
Tyr Ala Ala Pro Ala Pro Gly Gln Ala Leu Gln Arg Val Gly Ile Val
20 25 30
Gly Gly Gln Glu Ala Pro Arg Ser Lys Trp Pro Trp Gln Val Ser Leu
35 40 45
Arg Val Arg Asp Arg Tyr Trp Met His Phe Cys Gly Gly Ser Leu Ile
50 55 60
His Pro Gln Trp Val Leu Thr Ala Ala His Cys Val Gly Pro Asp Val
65 70 75 80
Lys Asp Leu Ala Ala Leu Arg ValGln Leu Arg Glu Gln His Leu Tyr
85 90 95
Tyr Gln Asp Gln Leu Leu Pro Val Ser Arg Ile Ile Val His Pro Gln
100 105 110
Phe Tyr Thr Ala Gln Ile Gly Ala Asp Ile Ala Leu Leu Glu Leu Glu
115 120 125
Glu Pro Val Asn Val Ser Ser His Val His Thr Val Thr Leu Pro Pro
130 135 140
Ala Ser Glu Thr Phe Pro Pro Gly Met Pro Cys Trp Val Thr Gly Trp
145 150 155 160
Gly Asp Val Asp Asn Asp Glu Arg Leu Pro Pro Pro Phe Pro Leu Lys
165 170 175
Gln Val Lys Val Pro Ile Met Glu Asn His Ile Cys Asp Ala Lys Tyr
180 185 190
His Leu Gly Ala Tyr Thr Gly Asp Asp Val Arg Ile Val Arg Asp Asp
195 200 205
Met Leu Cys Ala Gly Asn Thr Arg Arg Asp Ser Cys Gln Gly Asp Ser
210 215 220
Gly Gly Pro Leu Val Cys Lys Val Asn Gly Thr Trp Leu Gln Ala Gly
225 230 235 240
Val Val Ser Trp Gly Glu Gly Cys Ala Gln Pro Asn Arg Pro Gly Ile
245 250 255
Tyr Thr Arg Val Thr Tyr Tyr Leu Asp Trp Ile His His Tyr Val Pro
260 265 270
Lys Lys Pro
275
<210>212
<211>1013
<212>DNA
<213>Homo sapiens
<220>
<223>Neuropsin transcript variant 1
<400>212
gttcccagaa gctccccagg ctctagtgca ggaggagaag gaggaggagc aggaggtgga 60
gattcccagt taaaaggctc cagaatcgtg taccaggcag agaactgaag tactggggcc 120
tcctccactg ggtccgaatc agtaggtgac cccgcccctg gattctggaa gacctcacca 180
tgggacgccc ccgacctcgt gcggccaaga cgtggatgtt cctgctcttg ctggggggag 240
cctgggcagg acactccagg gcacaggagg acaaggtgct ggggggtcat gagtgccaac 300
cccattcgca gccttggcag gcggccttgt tccagggcca gcaactactc tgtggcggtg 360
tccttgtagg tggcaactgg gtccttacag ctgcccactg taaaaaaccg aaatacacag 420
tacgcctggg agaccacagc ctacagaata aagatggccc agagcaagaa atacctgtgg 480
ttcagtccat cccacacccc tgctacaaca gcagcgatgt ggaggaccac aaccatgatc 540
tgatgcttct tcaactgcgt gaccaggcat ccctggggtc caaagtgaag cccatcagcc 600
tggcagatca ttgcacccag cctggccaga agtgcaccgt ctcaggctgg ggcactgtca 660
ccagtccccg agagaatttt cctgacactc tcaactgtgc agaagtaaaa atctttcccc 720
agaagaagtg tgaggatgct tacccggggc agatcacaga tggcatggtc tgtgcaggca 780
gcagcaaagg ggctgacacg tgccagggcg attctggagg ccccctggtg tgtgatggtg 840
cactccaggg catcacatcc tggggctcag acccctgtgg gaggtccgac aaacctggcg 900
tctataccaa catctgccgc tacctggact ggatcaagaa gatcataggc agcaagggct 960
gattctagga taagcactag atctccctta ataaactcac aactctctgg ttc 1013
<210>213
<211>260
<212>PRT
<213>Homo sapiens
<220>
<223>Neuropsin isoform 1
<400>213
Met Gly Arg Pro Arg Pro Arg Ala Ala Lys Thr Trp Met Phe Leu Leu
1 5 10 15
Leu Leu Gly Gly Ala Trp Ala Gly His Ser Arg Ala Gln Glu Asp Lys
20 25 30
Val Leu Gly Gly His Glu Cys Gln Pro His Ser Gln Pro Trp Gln Ala
35 40 45
Ala Leu Phe Gln Gly Gln Gln Leu Leu Cys Gly Gly Val Leu Val Gly
50 55 60
Gly Asn Trp Val Leu Thr Ala Ala His Cys Lys Lys Pro Lys Tyr Thr
65 70 75 80
Val Arg Leu Gly Asp His Ser Leu Gln Asn Lys Asp Gly Pro Glu Gln
85 90 95
Glu Ile Pro Val Val Gln Ser Ile Pro His Pro Cys Tyr Asn Ser Ser
100 105 110
Asp Val Glu Asp His Asn His Asp Leu Met Leu Leu Gln Leu Arg Asp
115 120 125
Gln Ala Ser Leu Gly Ser Lys Val Lys Pro Ile Ser Leu Ala Asp His
130 135 140
Cys Thr Gln Pro Gly Gln Lys Cys Thr Val Ser Gly Trp Gly Thr Val
145 150 155 160
Thr Ser Pro Arg Glu Asn Phe Pro Asp Thr Leu Asn Cys Ala Glu Val
165 170 175
Lys Ile Phe Pro Gln Lys Lys Cys Glu Asp Ala Tyr Pro Gly Gln Ile
180 185 190
Thr Asp Gly Met Val Cys Ala Gly Ser Ser Lys Gly Ala Asp Thr Cys
195 200 205
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Asp Gly Ala Leu Gln Gly
210 215 220
Ile Thr Ser Trp Gly Ser Asp Pro Cys Gly Arg Ser Asp Lys Pro Gly
225 230 235 240
Val Tyr Thr Asn Ile Cys Arg Tyr Leu Asp Trp Ile Lys Lys Ile Ile
245 250 255
Gly Ser Lys Gly
260
<210>214
<211>1148
<212>DNA
<213>Homo sapiens
<220>
<223>Neuropsin transcript variant 2
<400>214
gttcccagaa gctccccagg ctctagtgca ggaggagaag gaggaggagc aggaggtgga 60
gattcccagt taaaaggctc cagaatcgtg taccaggcag agaactgaag tactggggcc 120
tcctccactg ggtccgaatc agtaggtgac cccgcccctg gattctggaa gacctcacca 180
tgggacgccc ccgacctcgt gcggccaaga cgtggatgtt cctgctcttg ctggggggag 240
cctgggcagc gtgtggaagc ctggacctcc tcactaagtt gtatgcggag aacttgccgt 300
gtgtccattt gaacccacag tggccttccc agccctcgca ctgccccaga gggtggcgat 360
ccaaccctct ccctcctgct gcaggacact ccagggcaca ggaggacaag gtgctggggg 420
gtcatgagtg ccaaccccat tcgcagcctt ggcaggcggc cttgttccag ggccagcaac 480
tactctgtgg cggtgtcctt gtaggtggca actgggtcct tacagctgcc cactgtaaaa 540
aaccgaaata cacagtacgc ctgggagacc acagcctaca gaataaagat ggcccagagc 600
aagaaatacc tgtggttcag tccatcccac acccctgcta caacagcagc gatgtggagg 660
accacaacca tgatctgatg cttcttcaac tgcgtgacca ggcatccctg gggtccaaag 720
tgaagcccat cagcctggca gatcattgca cccagcctgg ccagaagtgc accgtctcag 780
gctggggcac tgtcaccagt ccccgagaga attttcctga cactctcaac tgtgcagaag 840
taaaaatctt tccccagaag aagtgtgagg atgcttaccc ggggcagatc acagatggca 900
tggtctgtgc aggcagcagc aaaggggctg acacgtgcca gggcgattct ggaggccccc 960
tggtgtgtga tggtgcactc cagggcatca catcctgggg ctcagacccc tgtgggaggt 1020
ccgacaaacc tggcgtctat accaacatct gccgctacct ggactggatc aagaagatca 1080
taggcagcaa gggctgattc taggataagc actagatctc ccttaataaa ctcacaactc 1140
tctggttc 1148
<210>215
<211>305
<212>PRT
<213>Homo sapiens
<220>
<223>Neuropsin isoform 2
<400>215
Met Gly Arg Pro Arg Pro Arg Ala Ala Lys Thr Trp Met Phe Leu Leu
1 5 10 15
Leu Leu Gly Gly Ala Trp Ala Ala Cys Gly Ser Leu Asp Leu Leu Thr
20 25 30
Lys Leu Tyr Ala Glu Asn Leu Pro Cys Val His Leu Asn Pro Gln Trp
35 40 45
Pro Ser Gln Pro Ser His Cys Pro Arg Gly Trp Arg Ser Asn Pro Leu
50 55 60
Pro Pro Ala Ala Gly His Ser Arg Ala Gln Glu Asp Lys Val Leu Gly
65 70 75 80
Gly His Glu Cys Gln Pro His Ser Gln Pro Trp Gln Ala Ala Leu Phe
85 90 95
Gln Gly Gln Gln Leu Leu Cys Gly Gly Va1Leu Val Gly Gly Asn Trp
100 105 110
Val Leu Thr Ala Ala His Cys Lys Lys Pro Lys Tyr Thr Val Arg Leu
115 120 125
Gly Asp His Ser Leu Gln Asn Lys Asp Gly Pro Glu Gln Glu Ile Pro
130 135 140
Val Val Gln Ser Ile Pro His Pro Cys Tyr Asn Ser Ser Asp ValGlu
145 150 155 160
Asp His Asn His Asp Leu Met Leu Leu Gln Leu Arg Asp Gln Ala Ser
165 170 175
Leu Gly Ser Lys Val Lys Pro Ile Ser Leu Ala Asp His Cys Thr Gln
180 185 190
Pro Gly Gln Lys Cys Thr Val Ser Gly Trp Gly Thr Val Thr Ser Pro
195 200 205
Arg Glu Asn Phe Pro Asp Thr Leu Asn Cys Ala Glu ValLys Ile Phe
210 215 220
Pro Gln Lys Lys Cys Glu Asp Ala Tyr Pro Gly Gln Ile Thr Asp Gly
225 230 235 240
Met Val Cys Ala Gly Ser Ser Lys Gly Ala Asp Thr Cys Gln Gly Asp
245 250 255
Ser Gly Gly Pro Leu Val Cys Asp Gly Ala Leu Gln Gly Ile Thr Ser
260 265 270
Trp Gly Ser Asp Pro Cys Gly Arg Ser Asp Lys Pro Gly Val Tyr Thr
275 280 285
Asn Ile Cys Arg Tyr Leu Asp Trp Ile Lys Lys Ile Ile Gly Ser Lys
290 295 300
Gly
305
<210>216
<211>590
<212>DNA
<213>Homo sapiens
<220>
<223>Neuropsin transcript variant 3
<400>216
gttcccagaa gctccccagg ctctagtgca ggaggagaag gaggaggagc aggaggtgga 60
gattcccagt taaaaggctc cagaatcgtg taccaggcag agaactgaag tactggggcc 120
tcctccactg ggtccgaatc agtaggtgac cccgcccctg gattctggaa gacctcacca 180
tgggacgccc ccgacctcgt gcggccaaga cgtggatgtt cctgctcttg ctggggggag 240
cctgggcaga gaattttcct gacactctca actgtgcaga agtaaaaatc tttccccaga 300
agaagtgtga ggatgcttac ccggggcaga tcacagatgg catggtctgt gcaggcagca 360
gcaaaggggc tgacacgtgc cagggcgatt ctggaggccc cctggtgtgt gatggtgcac 420
tccagggcat cacatcctgg ggctcagacc cctgtgggag gtccgacaaa cctggcgtct 480
ataccaacat ctgccgctac ctggactgga tcaagaagat cataggcagc aagggctgat 540
tctaggataa gcactagatc tcccttaata aactcacaac tctctggttc 590
<210>217
<211>119
<212>PRT
<213>Homo sapiens
<220>
<223>Neuropsin isoform 3
<400>217
Met Gly Arg Pro Arg Pro Arg Ala Ala Lys Thr Trp Met Phe Leu Leu
1 5 10 15
Leu Leu Gly Gly Ala Trp Ala Glu Asn Phe Pro Asp Thr Leu Asn Cys
20 25 30
Ala Glu Val Lys Ile Phe Pro Gln Lys Lys Cys Glu Asp Ala Tyr Pro
35 40 45
Gly Gln Ile Thr Asp Gly Met Val Cys Ala Gly Ser Ser Lys Gly Ala
50 55 60
Asp Thr Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Asp Gly Ala
65 70 75 80
Leu Gln Gly Ile Thr Ser Trp Gly Ser Asp Pro Cys Gly Arg Ser Asp
85 90 95
Lys Pro Gly Val Tyr Thr Asn Ile Cys Arg Tyr Leu Asp Trp Ile Lys
100 105 110
Lys Ile Ile Gly Ser Lys Gly
115
<210>218
<211>456
<212>DNA
<213>Homo sapiens
<220>
<223>Neuropsin transcript variant 4
<400>218
gttcccagaa gctccccagg ctctagtgca ggaggagaag gaggaggagc aggaggtgga 60
gattcccagt taaaaggctc cagaatcgtg taccaggcag agaactgaag tactggggcc 120
tcctccactg ggtccgaatc agtaggtgac cccgcccctg gattctggaa gacctcacca 180
tgggacgccc ccgacctcgt gcggccaaga cgtggatgtt cctgctcttg ctggggggag 240
cctgggcagg gcgattctgg aggccccctg gtgtgtgatg gtgcactcca gggcatcaca 300
tcctggggct cagacccctg tgggaggtcc gacaaacctg gcgtctatac caacatctgc 360
cgctacctgg actggatcaa gaagatcata ggcagcaagg gctgattcta ggataagcac 420
tagatctccc ttaataaact cacaactctc tggttc 456
<210>219
<211>32
<212>PRT
<213>Homo sapiens
<220>
<223>Neuropsin isoform 4
<400>219
Met Gly Arg Pro Arg Pro Arg Ala Ala Lys Thr Trp Met Phe Leu Leu
1 5 10 15
Leu Leu Gly Gly Ala Trp Ala Gly Arg Phe Trp Arg Pro Pro Gly Val
20 25 30
<210>220
<211>1580
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK10 transcript variant 1
<400>220
catcctgcca cccctagcct tgctggggac gtgaaccctc tccccgcgcc tgggaagcct 60
tcttggcacc gggacccgga gaatccccac ggaagccagt tccaaaaggg atgaaaaggg 120
ggcgtttcgg gcactgggag aagcctgtat tccagggccc ctcccagagc aggaatctgg 180
gacccaggag tgccagcctc acccacgcag atcctggcca tgagagctcc gcacctccac 240
ctctccgccg cctctggcgc ccgggctctg gcgaagctgc tgccgctgct gatggcgcaa 300
ctctgggccg cagaggcggc gctgctcccc caaaacgaca cgcgcttgga ccccgaagcc 360
tatggctccc cgtgcgcgcg cggctcgcag ccctggcagg tctcgctctt caacggcctc 420
tcgttccact gcgcgggtgt cctggtggac cagagttggg tgctgacggc cgcgcactgc 480
ggaaacaagc cactgtgggc tcgagtaggg gatgaccacc tgctgcttct tcagggagag 540
cagctccgcc ggaccactcg ctctgttgtc catcccaagt accaccaggg ctcaggcccc 600
atcctgccaa ggcgaacgga tgagcacgat ctcatgttgc tgaagctggc caggcccgta 660
gtgctggggc cccgcgtccg ggccctgcag cttccctacc gctgtgctca gcccggagac 720
cagtgccagg ttgctggctg gggcaccacg gccgcccgga gagtgaagta caacaagggc 780
ctgacctgct ccagcatcac tatcctgagc cctaaagagt gtgaggtctt ctaccctggc 840
gtggtcacca acaacatgat atgtgctgga ctggaccggg gccaggaccc ttgccagagt 900
gactctggag gccccctggt ctgtgacgag accctccaag gcatcctctc gtggggtgtt 960
tacccctgtg gctctgccca gcatccagct gtctacaccc agatctgcaa atacatgtcc 1020
tggatcaata aagtcatacg ctccaactga tccagatgct acgctccagc tgatccagat 1080
gttatgctcc tgctgatcca gatgcccaga ggctccatcg tccatcctct tcctccccag 1140
tcggctgaac tctccccttg tctgcactgt tcaaacctct gccgccctcc acacctctaa 1200
acatctcccc tctcacctca ttcccccacc tatccccatt ctctgcctgt actgaagctg 1260
aaatgcagga agtggtggca aaggtttatt ccagagaagc caggaagccg gtcatcaccc 1320
agcctctgag agcagttact ggggtcaccc aacctgactt cctctgccac tccctgctgt 1380
gtgactttgg gcaagccaag tgccctctct gaacctcagt ttcctcatct gcaaaatggg 1440
aacaatgacg tgcctacctc ttagacatgt tgtgaggaga ctatgatata acatgtgtat 1500
gtaaatcttc atggtgattg tcatgtaagg cttaacacag tgggtggtga gttctgacta 1560
aaggttacct gttgtcgtga 1580
<210>221
<211>276
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hK10
<400>221
Met Arg Ala Pro His Leu His Leu Ser Ala Ala Ser Gly Ala Arg Ala
1 5 10 15
Leu Ala Lys Leu Leu Pro Leu Leu Met Ala Gln Leu Trp Ala Ala Glu
20 25 30
Ala Ala Leu Leu Pro Gln Asn Asp Thr Arg Leu Asp Pro Glu Ala Tyr
35 40 45
Gly Ser Pro Cys Ala Arg Gly Ser Gln Pro Trp Gln Val Ser Leu Phe
50 55 60
Asn Gly Leu Ser Phe His Cys Ala Gly Val Leu Val Asp Gln Ser Trp
65 70 75 80
Val Leu Thr Ala Ala His Cys Gly Asn Lys Pro Leu Trp Ala Arg Val
85 90 95
Gly Asp Asp His Leu Leu Leu Leu Gln Gly Glu Gln Leu Arg Arg Thr
100 105 110
Thr Arg Ser Val Val His Pro Lys Tyr His Gln Gly Ser Gly Pro Ile
115 120 125
Leu Pro Arg Arg Thr Asp Glu His Asp Leu Met Leu Leu Lys Leu Ala
130 135 140
Arg Pro Val Val Leu Gly Pro Arg Val Arg Ala Leu Gln Leu Pro Tyr
145 150 155 160
Arg Cys Ala Gln Pro Gly Asp Gln Cys Gln Val Ala Gly Trp Gly Thr
165 170 175
Thr Ala Ala Arg Arg Val Lys Tyr Asn Lys Gly Leu Thr Cys Ser Ser
180 185 190
Ile Thr Ile Leu Ser Pro Lys Glu Cys Glu Val Phe Tyr Pro Gly Val
195 200 205
Val Thr Asn Asn Met Ile Cys Ala Gly Leu Asp Arg Gly Gln Asp Pro
210 215 220
Cys Gln Ser Asp Ser Gly Gly Pro Leu Val Cys Asp Glu Thr Leu Gln
225 230 235 240
Gly Ile Leu Ser Trp Gly Val Tyr Pro Cys Gly Ser Ala Gln His Pro
245 250 255
Ala Val Tyr Thr Gln Ile Cys Lys Tyr Met Ser Trp Ile Asn Lys Val
260 265 270
Ile Arg Ser Asn
275
<210>222
<211>1443
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK10 transcript variant 2
<400>222
accagcggca gaccacaggc agggcagagg cacgtctggg tceectccct ccttcctate 60
ggcgactccc aggatcctgg ccatgagagc tccgcacctc cacctctccg ccgcctctgg 120
cgcccgggct ctggcgaagc tgctgccgct gctgatggcg caactctggg ccgcagaggc 180
ggcgctgctc ccccaaaacg acacgcgctt ggaccccgaa gcctatggct ccccgtgcgc 240
gcgcggctcg cagccctggc aggtctcgct cttcaacggc ctctcgttcc actgcgcggg 300
tgtcctggtg gaccagagtt gggtgctgac ggccgcgcac tgcggaaaca agccactgtg 360
ggctcgagta ggggatgacc acctgctgct tcttcaggga gagcagctcc gccggaccac 420
tcgctctgtt gtccatccca agtaccacca gggctcaggc cccatcctgc caaggcgaac 480
ggatgagcac gatctcatgt tgctgaagct ggccaggccc gtagtgctgg ggccccgcgt 540
ccgggccctg cagcttccct accgctgtgc tcagcccgga gaccagtgcc aggttgctgg 600
ctggggcacc acggccgccc ggagagtgaa gtacaacaag ggcctgacct gctccagcat 660
cactatcctg agccctaaag agtgtgaggt cttctaccct ggcgtggtca ccaacaacat 720
gatatgtgct ggactggacc ggggccagga cccttgccag agtgactctg gaggccccct 780
ggtctgtgac gagaccctcc aaggcatcct ctcgtggggt gtttacccct gtggctctgc 840
ccagcatcca gctgtctaca cccagatctg caaatacatg tcctggatca ataaagtcat 900
acgctccaac tgatccagat gctacgctcc agctgatcca gatgttatgc tcctgctgat 960
ccagatgccc agaggctcca tcgtccatcc tcttcctccc cagtcggctg aactctcccc 1020
ttgtctgcac tgttcaaacc tctgccgccc tccacacctc taaacatctc ccctctcacc 1080
tcattccccc acctatcccc attctctgcc tgtactgaag ctgaaatgca ggaagtggtg 1140
gcaaaggttt attccagaga agccaggaag ccggtcatca cccagcctct gagagcagtt 1200
actggggtca cccaacctga cttcctctgc cactccctgc tgtgtgactt tgggcaagcc 1260
aagtgccctc tctgaacctc agtttcctca tctgcaaaat gggaacaatg acgtgcctac 1320
ctcttagaca tgttgtgagg agactatgat ataacatgtg tatgtaaatc ttcatggtga 1380
ttgtcatgta aggcttaaca cagtgggtgg tgagttctga ctaaaggtta cctgttgtcg 1440
tga 1443
<210>223
<211>276
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hK10
<400>223
Met Arg Ala Pro His Leu His Leu Ser Ala Ala Ser Gly Ala Arg Ala
1 5 10 15
Leu Ala Lys Leu Leu Pro Leu Leu Met Ala Gln Leu Trp Ala Ala Glu
20 25 30
Ala Ala Leu Leu Pro Gln Asn Asp Thr Arg Leu Asp Pro Glu Ala Tyr
35 40 45
Gly Ser Pro Cys Ala Arg Gly Ser Gln Pro Trp Gln Val Ser Leu Phe
50 55 60
Asn Gly Leu Ser Phe His Cys Ala Gly Val Leu Val Asp Gln Ser Trp
65 70 75 80
Val Leu Thr Ala Ala His Cys Gly Asn Lys Pro Leu Trp Ala Arg Val
85 90 95
Gly Asp Asp His Leu Leu Leu Leu Gln Gly Glu Gln Leu Arg Arg Thr
100 105 110
Thr Arg Ser Val Val His Pro Lys Tyr His Gln Gly Ser Gly Pro Ile
115 120 125
Leu Pro Arg Arg Thr Asp Glu His Asp Leu Met Leu Leu Lys Leu Ala
130 135 140
Arg Pro Val Val Leu Gly Pro Arg Val Arg Ala Leu Gln Leu Pro Tyr
145 150 155 160
Arg Cys Ala Gln Pro Gly Asp Gln Cys Gln Val Ala Gly Trp Gly Thr
165 170 175
Thr Ala Ala Arg Arg Val Lys Tyr Asn Lys Gly Leu Thr Cys Ser Ser
180 185 190
Ile Thr Ile Leu Ser Pro Lys Glu Cys Glu Val Phe Tyr Pro Gly Val
195 200 205
Val Thr Asn Asn Met Ile Cys Ala Gly Leu Asp Arg Gly Gln Asp Pro
210 215 220
Cys Gln Ser Asp Ser Gly Gly Pro Leu Val Cys Asp Glu Thr Leu Gln
225 230 235 240
Gly Ile Leu Ser Trp Gly Val Tyr Pro Cys Gly Ser Ala Gln His Pro
245 250 255
Ala Val Tyr Thr Gln Ile Cys Lys Tyr Met Ser Trp Ile Asn Lys Val
260 265 270
Ile Arg Ser Asn
275
<210>224
<211>3226
<212>DNA
<213>Homo sapiens
<220>
<223>Epitheliasin
<400>224
cgcgagctaa gcaggaggcg gaggcggagg cggagggcga ggggcgggga gcgccgcctg 60
gagcgcggca ggtcatattg aacattccag atacctatca ttactcgatg ctgttgataa 120
cagcaagatg gctttgaact cagggtcacc accagctatt ggaccttact atgaaaacca 180
tggataccaa ccggaaaacc cctatcccgc acagcccact gtggtcccca ctgtctacga 240
ggtgcatccg gctcagtact acccgtcccc cgtgccccag tacgccccga gggtcctgac 300
gcaggcttcc aaccccgtcg tctgcacgca gcccaaatcc ccatccggga cagtgtgcac 360
ctcaaagact aagaaagcac tgtgcatcac cttgaccctg gggaccttcc tcgtgggagc 420
tgcgctggcc gctggcctac tctggaagtt catgggcagc aagtgctcca actctgggat 480
agagtgcgac tcctcaggta cctgcatcaa cccctctaac tggtgtgatg gcgtgtcaca 540
ctgccccggc ggggaggacg agaatcggtg tgttcgcctc tacggaccaa acttcatcct 600
tcagatgtac tcatctcaga ggaagtcctg gcaccctgtg tgccaagacg actggaacga 660
gaactacggg cgggcggcct gcagggacat gggctataag aataattttt actctagcca 720
aggaatagtg gatgacagcg gatccaccag ctttatgaaa ctgaacacaa gtgccggcaa 780
tgtcgatatc tataaaaaac tgtaccacag tgatgcctgt tcttcaaaag cagtggtttc 840
tttacgctgt atagcctgcg gggtcaactt gaactcaagc cgccagagca ggatcgtggg 900
cggtgagagc gcgctcccgg gggcctggcc ctggcaggtc agcctgcacg tccagaacgt 960
ccacgtgtgc ggaggctcca tcatcacccc cgagtggatc gtgacagccg cccactgcgt 1020
ggaaaaacct cttaacaatc catggcattg gacggcattt gcggggattt tgagacaatc 1080
tttcatgttc tatggagccg gataccaagt agaaaaagtg atttctcatc caaattatga 1140
ctccaagacc aagaacaatg acattgcgct gatgaagctg cagaagcctc tgactttcaa 1200
cgacctagtg aaaccagtgt gtctgcccaa cccaggcatg atgctgcagc cagaacagct 1260
ctgctggatt tccgggtggg gggccaccga ggagaaaggg aagacctcag aagtgctgaa 1320
cgctgccaag gtgcttctca ttgagacaca gagatgcaac agcagatatg tctatgacaa 1380
cctgatcaca ccagccatga tctgtgccgg cttcctgcag gggaacgtcg attcttgcca 1440
gggtgacagt ggagggcctc tggtcacttc gaagaacaat atctggtggc tgatagggga 1500
tacaagctgg ggttctggct gtgccaaagc ttacagacca ggagtgtacg ggaatgtgat 1560
ggtattcacg gactggattt atcgacaaat gagggcagac ggctaatcca catggtcttc 1620
gtccttgacg tcgttttaca agaaaacaat ggggctggtt ttgcttcccc gtgcatgatt 1680
tactcttaga gatgattcag aggtcacttc atttttatta aacagtgaac ttgtctggct 1740
ttggcactct ctgccattct gtgcaggctg cagtggctcc cctgcccagc ctgctctccc 1800
taaccccttg tccgcaaggg gtgatggccg gctggttgtg ggcactggcg gtcaagtgtg 1860
gaggagaggg gtggaggctg ccccattgag atcttcctgc tgagtccttt ccaggggcca 1920
attttggatg agcatggagc tgtcacctct cagctgctgg atgacttgag atgaaaaagg 1980
agagacatgg aaagggagac agccaggtgg cacctgcagc ggctgccctc tggggccact 2040
tggtagtgtc cccagcctac ctctccacaa ggggattttg ctgatgggtt cttagagcct 2100
tagcagccct ggatggtggc cagaaataaa gggaccagcc cttcatgggt ggtgacgtgg 2160
tagtcacttg taaggggaac agaaacattt ttgttcttat ggggtgagaa tatagacagt 2220
gcccttggtg cgagggaagc aattgaaaag gaacttgccc tgagcactcc tggtgcaggt 2280
ctccacctgc acattgggtg gggctcctgg gagggagact cagccttcct cctcatcctc 2340
cctgaccctg ctcctagcac cctggagagt gcacatgccc cttggtcctg gcagggcgcc 2400
aagtctggca ccatgttggc ctcttcaggc ctgctagtca ctggaaattg aggtccatgg 2460
gggaaatcaa ggatgctcag tttaaggtac actgtttcca tgttatgttt ctacacattg 2520
ctacctcagt gctcctggaa acttagcttt tgatgtctcc aagtagtcca ccttcattta 2580
actctttgaa actgtatcac ctttgccaag taagagtggt ggcctatttc agctgctttg 2640
acaaaatgac tggctcctga cttaacgttc tataaatgaa tgtgctgaag caaagtgccc 2700
atggtggcgg cgaagaagag aaagatgtgt tttgttttgg actctctgtg gtcccttcca 2760
atgctgtggg tttccaacca ggggaagggt cccttttgca ttgccaagtg ccataaccat 2820
gagcactact ctaccatggt tctgcctcct ggccaagcag gctggtttgc aagaatgaaa 2880
tgaatgattc tacagctagg acttaacctt gaaatggaaa gtcttgcaat cccatttgca 2940
ggatccgtct gtgcacatgc ctctgtagag agcagcattc ccagggacct tggaaacagt 3000
tggcactgta aggtgcttgc tccccaagac acatcctaaa aggtgttgta atggtgaaaa 3060
cgtcttcctt ctttattgcc ccttcttatt tatgtgaaca actgtttgtc tttttttgta 3120
tcttttttaa actgtaaagt tcaattgtga aaatgaatat catgcaaata aattatgcga 3180
tttttttttc aaagcaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaa 3226
<210>225
<211>492
<212>PRT
<213>Homo sapiens
<220>
<223>Epitheliasin
<400>225
Met Ala Leu Asn Ser Gly Ser Pro Pro Ala Ile Gly Pro Tyr Tyr Glu
1 5 10 15
Asn His Gly Tyr Gln Pro Glu Asn Pro Tyr Pro Ala Gln Pro Thr Val
20 25 30
Val Pro Thr Val Tyr Glu Val His Pro Ala Gln Tyr Tyr Pro Ser Pro
35 40 45
Val Pro Gln Tyr Ala Pro Arg Val Leu Thr Gln Ala Ser Asn Pro Val
50 55 60
Val Cys Thr Gln Pro Lys Ser Pro Ser Gly Thr Val Cys Thr Ser Lys
65 70 75 80
Thr Lys Lys Ala Leu Cys Ile Thr Leu Thr Leu Gly Thr Phe Leu Val
85 90 95
Gly Ala Ala Leu Ala Ala Gly Leu Leu Trp Lys Phe Met Gly Ser Lys
100 105 110
Cys Ser Asn Ser Gly Ile Glu Cys Asp Ser Ser Gly Thr Cys Ile Asn
115 120 125
Pro Ser Asn Trp Cys Asp Gly Val Ser His Cys Pro Gly Gly Glu Asp
130 135 140
Glu Asn Arg Cys ValArg Leu Tyr Gly Pro Asn Phe Ile Leu Gln Met
145 150 155 160
Tyr Ser Ser Gln Arg Lys Ser Trp His Pro Val Cys Gln Asp Asp Trp
165 170 175
Asn Glu Asn Tyr Gly Arg Ala Ala Cys Arg Asp Met Gly Tyr Lys Asn
180 185 190
Asn Phe Tyr Ser Ser Gln Gly Ile Val Asp Asp Ser Gly Ser Thr Ser
195 200 205
Phe Met Lys Leu Asn Thr Ser Ala Gly Asn Val Asp Ile Tyr Lys Lys
210 215 220
Leu Tyr His Ser Asp Ala Cys Ser Ser Lys Ala Val Val Ser Leu Arg
225 230 235 240
Cys Ile Ala Cys Gly Val Asn Leu Asn Ser Ser Arg Gln Ser Arg Ile
245 250 255
Val Gly Gly Glu Ser Ala Leu Pro Gly Ala Trp Pro Trp Gln Val Ser
260 265 270
Leu His Val Gln Asn Val His Val Cys Gly Gly Ser Ile Ile Thr Pro
275 280 285
Glu Trp Ile Val Thr Ala Ala His Cys Val Glu Lys Pro Leu Asn Asn
290 295 300
Pro Trp His Trp Thr Ala Phe Ala Gly Ile Leu Arg Gln Ser Phe Met
305 310 315 320
Phe Tyr Gly Ala Gly Tyr Gln Val Glu Lys Val Ile Ser His Pro Asn
325 330 335
Tyr Asp Ser Lys Thr Lys Asn Asn Asp Ile Ala Leu Met Lys Leu Gln
340 345 350
Lys Pro Leu Thr Phe Asn Asp Leu ValLys Pro Val Cys Leu Pro Asn
355 360 365
Pro Gly Met Met Leu Gln Pro Glu Gln Leu Cys Trp Ile Ser Gly Trp
370 375 380
Gly Ala Thr Glu Glu Lys Gly Lys Thr Ser Glu Val Leu Asn Ala Ala
385 390 395 400
Lys Val Leu Leu Ile Glu Thr Gln Arg Cys Asn Ser Arg Tyr Val Tyr
405 410 415
Asp Asn Leu Ile Thr Pro Ala Met Ile Cys Ala Gly Phe Leu Gln Gly
420 425 430
Asn Val Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Thr Ser
435 440 445
Lys Asn Asn Ile Trp Trp Leu Ile Gly Asp Thr Ser Trp Gly Ser Gly
450 455 460
Cys Ala Lys Ala Tyr Arg Pro Gly Val Tyr Gly Asn Val Met Val Phe
465 470 475 480
Thr Asp Trp Ile Tyr Arg Gln Met Arg Ala Asp Gly
485 490
<210>226
<211>1280
<212>DNA
<213>Homo sapiens
<220>
<223>Prostase
<400>226
atggccacag caggaaatcc ctggggctgg ttcctggggt acctcatcct tggtgtcgca 60
ggatctctcg tctctggtag ctgcagccaa atcataaacg gcgaggactg cagcccgcac 120
tcgcagccct ggcaggcggc actggtcatg gaaaacgaat tgttctgctc gggcgtcctg 180
gtgcatccgc agtgggtgct gtcagccgca cactgtttcc agaactccta caccatcggg 240
ctgggcctgc acagtcttga ggccgaccaa gagccaggga gccagatggt ggaggccagc 300
ctctccgtac ggcacccaga gtacaacaga cccttgctcg ctaacgacct catgctcatc 360
aagttggacg aatccgtgtc cgagtctgac accatccgga gcatcagcat tgcttcgcag 420
tgccctaccg cggggaactc ttgcctcgtt tctggctggg gtctgctggc gaacggcaga 480
atgcctaccg tgctgcagtg cgtgaacgtg tcggtggtgt ctgaggaggt ctgcagtaag 540
ctctatgacc cgctgtacca ccccagcatg ttctgcgccg gcggagggca agaccagaag 600
gactcctgca acggtgactc tggggggccc ctgatctgca acgggtactt gcagggcctt 660
gtgtctttcg gaaaagcccc gtgtggccaa gttggcgtgc caggtgtcta caccaacctc 720
tgcaaattca ctgagtggat agagaaaacc gtccaggcca gttaactctg gggactggga 780
acccatgaaa ttgaccccca aatacatcct gcggaaggaa ttcaggaata tctgttccca 840
gcccctcctc cctcaggccc aggagtccag gcccccagcc cctcctccct caaaccaagg 900
gtacagatcc ccagcccctc ctccctcaga cccaggagtc cagacccccc agcccctcct 960
ccctcagacc caggagtcca gcccctcctc cctcagaccc aggagtccag accccccagc 1020
ccctcctccc tcagacccag gggtccaggc ccccaacccc tcctccctca gactcagagg 1080
tccaagcccc caacccctcc ttccccagac ccagaggtcc aggtcccagc ccctcctccc 1140
tcagacccag cggtccaatg ccacctagac tctccctgta cacagtgccc ccttgtggca 1200
cgttgaccca accttaccag ttggtttttc attttttgtc cctttcccct agatccagaa 1260
ataaagtcta agagaagcgc 1280
<210>227
<211>254
<212>PRT
<213>Homo sapiens
<220>
<223>Prostase
<400>227
Met Ala Thr Ala Gly Asn Pro Trp Gly Trp Phe Leu Gly Tyr Leu Ile
1 5 10 15
Leu Gly Val Ala Gly Ser Leu Val Ser Gly Ser Cys Ser Gln Ile Ile
20 25 30
Asn Gly Glu Asp Cys Ser Pro His Ser Gln Pro Trp Gln Ala Ala Leu
35 40 45
Val Met Glu Asn Glu Leu Phe Cys Ser Gly Val Leu Val His Pro Gln
50 55 60
Trp Val Leu Ser Ala Ala His Cys Phe Gln Asn Ser Tyr Thr Ile Gly
65 70 75 80
Leu Gly Leu His Ser Leu Glu Ala Asp Gln Glu Pro Gly Ser Gln Met
85 90 95
Val Glu Ala Ser Leu Ser Val Arg His Pro Glu Tyr Asn Arg Pro Leu
100 105 110
Leu Ala Asn Asp Leu Met Leu Ile Lys Leu Asp Glu Ser Val Ser Glu
115 120 125
Ser Asp Thr Ile Arg Ser Ile Ser Ile Ala Ser Gln Cys Pro Thr Ala
130 135 140
Gly Asn Ser Cys Leu Val Ser Gly Trp Gly Leu Leu Ala Asn Gly Arg
145 150 155 160
Met Pro Thr Val Leu Gln Cys Val Asn Val Ser Val Val Ser Glu Glu
165 170 175
Val Cys Ser Lys Leu Tyr Asp Pro Leu Tyr His Pro Ser Met Phe Cys
180 185 190
Ala Gly Gly Gly Gln Asp Gln Lys Asp Ser Cys Asn Gly Asp Ser Gly
195 200 205
Gly Pro Leu Ile Cys Asn Gly Tyr Leu Gln Gly Leu Val Ser Phe Gly
210 215 220
Lys Ala Pro Cys Gly Gln Val Gly Val Pro Gly Val Tyr Thr Asn Leu
225 230 235 240
Cys Lys Phe Thr Glu Trp Ile Glu Lys Thr Val Gln Ala Ser
245 250
<210>228
<211>1391
<212>DNA
<213>Homo sapiens
<220>
<223>Brain serine proteinase 2
<400>228
acacaccctg acccgcatcg ccctgggtct ctcgagcctg ctgcctgctc ccccgcccca 60
ccagccatgg tggtttctgg agcgccccca gccctgggtg ggggctgtct cggcaccttc 120
acctccctgc tgctgctggc gtcgacagcc atcctcaatg cggccaggat acctgttccc 180
ccagcctgtg ggaagcccca gcagctgaac cgggttgtgg gcggcgagga cagcactgac 240
agcgagtggc cctggatcgt gagcatccag aagaatggga cccaccactg cgcaggttct 300
ctgctcacca gccgctgggt gatcactgct gcccactgtt tcaaggacaa cctgaacaaa 360
ccatacctgt tctctgtgct gctgggggcc tggcagctgg ggaaccctgg ctctcggtcc 420
cagaaggtgg gtgttgcctg ggtggagccc caccctgtgt attcctggaa ggaaggtgcc 480
tgtgcagaca ttgccctggt gcgtctcgag cgctccatac agttctcaga gcgggtcctg 540
cccatctgcc tacctgatgc ctctatccac ctccctccaa acacccactg ctggatctca 600
ggctggggga gcatccaaga tggagttccc ttgccccacc ctcagaccct gcagaagctg 660
aaggttccta tcatcgactc ggaagtctgc agccatctgt actggcgggg agcaggacag 720
ggacccatca ctgaggacat gctgtgtgcc ggctacttgg agggggagcg ggatgcttgt 780
ctgggcgact ccgggggccc cctcatgtgc caggtggacg gcgcctggct gctggccggc 840
atcatcagct ggggcgaggg ctgtgccgag cgcaacaggc ccggggtcta catcagcctc 900
tctgcgcacc gctcctgggt ggagaagatc gtgcaagggg tgcagctccg cgggcgcgct 960
caggggggtg gggccctcag ggcaccgagc cagggctctg gggccgccgc gcgctcctag 1020
ggcgcagcgg gacgcggggc tcggatctga aaggcggcca gatccacatc tggatctgga 1080
tctgcggcgg cctcgggcgg tttcccccgc cgtaaatagg ctcatctacc tctacctctg 1140
ggggcccgga cggctgctgc ggaaaggaaa ccccctcccc gacccgcccg acggcctcag 1200
gccccgcctc caaggcatca ggccccgccc aacggcctca tgtccccgcc cccacgactt 1260
ccggccccgc cccgggcccc agcgcttttg tgtatataaa tgttaatgat ttttataggt 1320
atttgtaacc ctgcccacat atcttattta ttcctccaat ttcaataaat tatttattct 1380
ccagtgaaaa a 1391
<210>229
<211>317
<212>PRT
<213>Homo sapiens
<220>
<223>Brain serine proteinase 2
<400>229
Met Val Val Ser Gly Ala Pro Pro Ala Leu Gly Gly Gly Cys Leu Gly
1 5 10 15
Thr Phe Thr Ser Leu Leu Leu Leu Ala Ser Thr Ala Ile Leu Asn Ala
20 25 30
Ala Arg Ile Pro Val Pro Pro Ala Cys Gly Lys Pro Gln Gln Leu Asn
35 40 45
Arg Val Val Gly Gly Glu Asp Ser Thr Asp Ser Glu Trp Pro Trp Ile
50 55 60
Val Ser Ile Gln Lys Asn Gly Thr His His Cys Ala Gly Ser Leu Leu
65 70 75 80
Thr Ser Arg Trp Val Ile Thr Ala Ala His Cys Phe Lys Asp Asn Leu
85 90 95
Asn Lys Pro Tyr Leu Phe Ser Val Leu Leu Gly Ala Trp Gln Leu Gly
100 105 110
Asn Pro Gly Ser Arg Ser Gln Lys Val Gly Val Ala Trp Val Glu Pro
115 120 125
His Pro Val Tyr Ser Trp Lys Glu Gly Ala Cys Ala Asp Ile Ala Leu
130 135 140
Val Arg Leu Glu Arg Ser Ile Gln Phe Ser Glu Arg Val Leu Pro Ile
145 150 155 160
Cys Leu Pro Asp Ala Ser Ile His Leu Pro Pro Asn Thr His Cys Trp
165 170 175
Ile Ser Gly Trp Gly Ser Ile Gln Asp Gly Val Pro Leu Pro His Pro
180 185 190
Gln Thr Leu Gln Lys Leu Lys Val Pro Ile Ile Asp Ser Glu Val Cys
195 200 205
Ser His Leu Tyr Trp Arg Gly Ala Gly Gln Gly Pro Ile Thr Glu Asp
210 215 220
Met Leu Cys Ala Gly Tyr Leu Glu Gly Glu Arg Asp Ala Cys Leu Gly
225 230 235 240
Asp Ser Gly Gly Pro Leu Met Cys Gln Val Asp Gly Ala Trp Leu Leu
245 250 255
Ala Gly Ile Ile Ser Trp Gly Glu Gly Cys Ala Glu Arg Asn Arg Pro
260 265 270
Gly Val Tyr Ile Ser Leu Ser Ala His Arg Ser Trp Val Glu Lys Ile
275 280 285
Val Gln Gly Val Gln Leu Arg Gly Arg Ala Gln Gly Gly Gly Ala Leu
290 295 300
Arg Ala Pro Ser Gln Gly Ser Gly Ala Ala Ala Arg Ser
305 310 315
<210>230
<211>1184
<212>DNA
<213>Homo sapiens
<220>
<223>Chymopasin
<400>230
atctgccacg atgttgctgc tcagcctgac cctaagcctg gttctcctcg gctcctcctg 60
gggctgcggc attcctgcca tcaaaccggc actgagcttc agccagagga ttgtcaacgg 120
ggagaatgca gtgttgggct cctggccctg gcaggtgtcc ctgcaggaca gcagcggctt 180
ccacttctgc ggtggttctc tcatcagcca gtcctgggtg gtcactgctg cccactgcaa 240
tgtcagccct ggccgccatt ttgttgtcct gggcgagtat gaccgatcat caaacgcaga 300
gcccttgcag gttctgtccg tctctcgggc cattacacac cctagctgga actctaccac 360
catgaacaat gacgtgacgc tgctgaagct cgcctcgcca gcccagtaca caacacgcat 420
ctcgccagtt tgcctggcat cctcaaacga ggctctgact gaaggcctca cgtgtgtcac 480
caccggctgg ggtcgcctca gtggcgtggg caatgtgaca ccagcacatc tgcagcaggt 540
ggctttgccc ctggtcactg tgaatcagtg ccggcagtac tggggctcaa gtatcactga 600
ctccatgatc tgtgcaggtg gcgcaggtgc ctcctcgtgc cagggtgact ccggaggccc 660
tcttgtctgc cagaagggaa acacatgggt gcttattggt attgtctcct ggggcaccaa 720
aaactgcaat gtgcgcgcac ctgctgtgta tactcgagtt agcaagttca gcacctggat 780
caaccaggtc atagcctaca actgagctca ccacaggccc tccccagctc aacccattaa 840
agacccaggc cctgtcccat catgcattca tgtctgtctt cctggctcag gagaaagaag 900
aggctgttga gggtccgact ccctacttgg acttctggca cagaaggggc tgagtgactc 960
cttgagtagc agtggctctt cctagagtag ccatgccgag gccggggccc ccacccctcc 1020
tccagggcaa ccccttggtc ctacagcaag aagccagaac tgttggaatg aatggcagcc 1080
ctccctggag aggcagcctg tttactgaat acagaggata cgtttacaaa ctgaatacgc 1140
ataataaata actgcacatt ctccatccaa aaaaaaaaaa aaaa 1184
<210>231
<211>264
<212>PRT
<213>Homo sapiens
<220>
<223>Chymopasin
<400>231
Met Leu Leu Leu Ser Leu Thr Leu Ser Leu Val Leu Leu Gly Ser Ser
1 5 10 15
Trp Gly Cys Gly Ile Pro Ala Ile Lys Pro Ala Leu Ser Phe Ser Gln
20 25 30
Arg Ile Val Asn Gly Glu Asn Ala Val Leu Gly Ser Trp Pro Trp Gln
35 40 45
Val Ser Leu Gln Asp Ser Ser Gly Phe His Phe Cys Gly Gly Ser Leu
50 55 60
Ile Ser Gln Ser Trp Val Val Thr Ala Ala His Cys Asn Val Ser Pro
65 70 75 80
Gly Arg His Phe Val Val Leu Gly Glu Tyr Asp Arg Ser Ser Asn Ala
85 90 95
Glu Pro Leu Gln Val Leu Ser Val Ser Arg Ala Ile Thr His Pro Ser
100 105 110
Trp Asn Ser Thr Thr Met Asn Asn Asp Val Thr Leu Leu Lys Leu Ala
115 120 125
Ser Pro Ala Gln Tyr Thr Thr Arg Ile Ser Pro Val Cys Leu Ala Ser
130 135 140
Ser Asn Glu Ala Leu Thr Glu Gly Leu Thr Cys Val Thr Thr Gly Trp
145 150 155 160
Gly Arg Leu Ser Gly Val Gly Asn Val Thr Pro Ala His Leu Gln Gln
165 170 175
Val Ala Leu Pro Leu Val Thr ValAsn Gln Cys Arg Gln Tyr Trp Gly
180 185 190
Ser Ser Ile Thr Asp Ser Met Ile Cys Ala Gly Gly Ala Gly Ala Ser
195 200 205
Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Gln Lys Gly Asn
210 215 220
Thr Trp Val Leu Ile Gly Ile Val Ser Trp Gly Thr Lys Asn Cys Asn
225 230 235 240
Val Arg Ala Pro Ala Val Tyr Thr Arg Val Ser Lys Phe Ser Thr Trp
245 250 255
Ile Asn Gln Val Ile Ala Tyr Asn
260
<210>232
<211>1207
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein 11 transcript variant 1
<400>232
ggagtgcagg cttggaaagc aggagagctc agcctacgtc tttaatcctc ctgcccaccc 60
cttggattct gtctccactg ggactcaaga gaggaacctg gggcccgctc ctcccccctc 120
caggccatga ggattctgca gttaatcctg cttgctctgg caacagggct tgtaggggga 180
gagaccagga tcatcaaggg gttcgagtgc aagcctcact cccagccctg gcaggcagcc 240
ctgttcgaga agacgcggct actctgtggg gcgacgctca tcgcccccag atggctcctg 300
acagcagccc actgcctcaa gccccgctac atagttcacc tggggcagca caacctccag 360
aaggaggagg gctgtgagca gacccggaca gccactgagt ccttccccca ccccggcttc 420
aacaacagcc tccccaacaa agaccaccgc aatgacatca tgctggtgaa gatggcatcg 480
ccagtctcca tcacctgggc tgtgcgaccc ctcaccctct cctcacgctg tgtcactgct 540
ggcaccagct gcctcatttc cggctggggc agcacgtcca gcccccagtt acgcctgcct 600
cacaccttgc gatgcgccaa catcaccatc attgagcacc agaagtgtga gaacgcctac 660
cccggcaaca tcacagacac catggtgtgt gccagcgtgc aggaaggggg caaggactcc 720
tgccagggtg actccggggg ccctctggtc tgtaaccagt ctcttcaagg cattatctcc 780
tggggccagg atccgtgtgc gatcacccga aagcctggtg tctacacgaa agtctgcaaa 840
tatgtggact ggatccagga gacgatgaag aacaattaga ctggacccac ccaccacagc 900
ccatcaccct ccatttccac ttggtgtttg gttcctgttc actctgttaa taagaaaccc 960
taagccaaga ccctctacga acattctttg ggcctcctgg actacaggag atgctgtcac 1020
ttaataatca acctggggtt cgaaatcagt gagacctgga ttcaaattct gccttgaaat 1080
attgtgactc tgggaatgac aacacctggt ttgttctctg ttgtatcccc agccccaaag 1140
acagctcctg gccatatatc aaggtttcaa taaatatttg ctaaatgagt gaaaaaaaaa 1200
aaaaaaa 1207
<210>233
<211>250
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein 11 isoform 1
<400>233
Met Arg Ile Leu Gln Leu Ile Leu Leu Ala Leu Ala Thr Gly Leu Val
1 5 10 15
Gly Gly Glu Thr Arg Ile Ile Lys Gly Phe Glu Cys Lys Pro His Ser
20 25 30
Gln Pro Trp Gln Ala Ala Leu Phe Glu Lys Thr Arg Leu Leu Cys Gly
35 40 45
Ala Thr Leu Ile Ala Pro Arg Trp Leu Leu Thr Ala Ala His Cys Leu
50 55 60
Lys Pro Arg Tyr Ile Val His Leu Gly Gln His Asn Leu Gln Lys Glu
65 70 75 80
Glu Gly Cys Glu Gln Thr Arg Thr Ala Thr Glu Ser Phe Pro His Pro
85 90 95
Gly Phe Asn Asn Ser Leu Pro Asn Lys Asp His Arg Asn Asp Ile Met
100 105 110
Leu Val Lys Met Ala Ser Pro Val Ser Ile Thr Trp Ala Val Arg Pro
115 120 125
Leu Thr Leu Ser Ser Arg Cys Val Thr Ala Gly Thr Ser Cys Leu Ile
130 135 140
Ser Gly Trp Gly Ser Thr Ser Ser Pro Gln Leu Arg Leu Pro His Thr
145 150 155 160
Leu Arg Cys Ala Asn Ile Thr Ile Ile Glu His Gln Lys Cys Glu Asn
165 170 175
Ala Tyr Pro Gly Asn Ile Thr Asp Thr Met Val Cys Ala Ser Val Gln
180 185 190
Glu Gly Gly Lys Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Val
195 200 205
Cys Asn Gln Ser Leu Gln Gly Ile Ile Ser Trp Gly Gln Asp Pro Cys
210 215 220
Ala Ile Thr Arg Lys Pro Gly Val Tyr Thr Lys Val Cys Lys Tyr Val
225 230 235 240
Asp Trp Ile Gln Glu Thr Met Lys Asn Asn
245 250
<210>234
<211>1289
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein 11 transcript variant 2
<400>234
ctgccttgct ccacacctgg tcaggggaga gaggggagga aagccaaggg aagggaccta 60
actgaaaaca aacaagctgg gagaagcagg aatctgcgct cgggttccgc agatgcagag 120
gttgaggtgg ctgcgggact ggaagtcatc gggcagaggt ctcacagcag ccaaggaacc 180
tggggcccgc tcctcccccc tccaggccat gaggattctg cagttaatcc tgcttgctct 240
ggcaacaggg cttgtagggg gagagaccag gatcatcaag gggttcgagt gcaagcctca 300
ctcccagccc tggcaggcag ccctgttcga gaagacgcgg ctactctgtg gggcgacgct 360
catcgccccc agatggctcc tgacagcagc ccactgcctc aagccccgct acatagttca 420
cctggggcag cacaacctcc agaaggagga gggctgtgag cagacccgga cagccactga 480
gtccttcccc caccccggct tcaacaacag cctccccaac aaagaccacc gcaatgacat 540
catgctggtg aagatggcat cgccagtctc catcacctgg gctgtgcgac ccctcaccct 600
ctcctcacgc tgtgtcactg ctggcaccag ctgcctcatt tccggctggg gcagcacgtc 660
cagcccccag ttacgcctgc ctcacacctt gcgatgcgcc aacatcacca tcattgagca 720
ccagaagtgt gagaacgcct accccggcaa catcacagac accatggtgt gtgccagcgt 780
gcaggaaggg ggcaaggact cctgccaggg tgactccggg ggccctctgg tctgtaacca 840
gtctcttcaa ggcattatct cctggggcca ggatccgtgt gcgatcaccc gaaagcctgg 900
tgtctacacg aaagtctgca aatatgtgga ctggatccag gagacgatga agaacaatta 960
gactggaccc acccaccaca gcccatcacc ctccatttcc acttggtgtt tggttcctgt 1020
tcactctgtt aataagaaac cctaagccaa gaccctctac gaacattctt tgggcctcct 1080
ggactacagg agatgctgtc acttaataat caacctgggg ttcgaaatca gtgagacctg 1140
gattcaaatt ctgccttgaa atattgtgac tctgggaatg acaacacctg gtttgttctc 1200
tgttgtatcc ccagccccaa agacagctcc tggccatata tcaaggtttc aataaatatt 1260
tgctaaatga gtgaaaaaaa aaaaaaaaa 1289
<210>235
<211>282
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein 11 isoform 2
<400>235
Met Gln Arg Leu Arg Trp Leu Arg Asp Trp Lys Ser Ser Gly Arg Gly
1 5 10 15
Leu Thr Ala Ala Lys Glu Pro Gly Ala Arg Ser Ser Pro Leu Gln Ala
20 25 30
Met Arg Ile Leu Gln Leu Ile Leu Leu Ala Leu Ala Thr Gly Leu Val
35 40 45
Gly Gly Glu Thr Arg Ile Ile Lys Gly Phe Glu Cys Lys Pro His Ser
50 55 60
Gln Pro Trp Gln Ala Ala Leu Phe Glu Lys Thr Arg Leu Leu Cys Gly
65 70 75 80
Ala Thr Leu Ile Ala Pro Arg Trp Leu Leu Thr Ala Ala His Cys Leu
85 90 95
Lys Pro Arg Tyr Ile Val His Leu Gly Gln His Asn Leu Gln Lys Glu
100 105 110
Glu Gly Cys Glu Gln Thr Arg Thr Ala Thr Glu Ser Phe Pro His Pro
115 120 125
Gly Phe Asn Asn Ser Leu Pro Asn Lys Asp His Arg Asn Asp Ile Met
130 135 140
Leu Val Lys Met Ala Ser Pro Val Ser Ile Thr Trp Ala Val Arg Pro
145 150 155 160
Leu Thr Leu Ser Ser Arg Cys Val Thr Ala Gly Thr Ser Cys Leu Ile
165 170 175
Ser Gly Trp Gly Ser Thr Ser Ser Pro Gln Leu Arg Leu Pro His Thr
180 185 190
Leu Arg Cys Ala Asn Ile Thr Ile Ile Glu His Gln Lys Cys Glu Asn
195 200 205
Ala Tyr Pro Gly Asn Ile Thr Asp Thr Met Val Cys Ala Ser Val Gln
210 215 220
Glu Gly Gly Lys Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Val
225 230 235 240
Cys Asn Gln Ser Leu Gln Gly Ile Ile Ser Trp Gly Gln Asp Pro Cys
245 250 255
Ala Ile Thr Arg Lys Pro Gly Val Tyr Thr Lys Val Cys Lys Tyr Val
260 265 270
Asp Trp Ile Gln Glu Thr Met Lys Asn Asn
275 280
<210>236
<211>802
<212>DNA
<213>Homo sapiens
<220>
<223>anionic trypsin (II)
<400>236
accaccatga atctacttct gatccttacc tttgttgcag ctgctgttgc tgcccccttt 60
gatgatgatg acaagatcgt tgggggctac atctgtgagg agaattctgt cccctaccag 120
gtgtccttga attctggcta ccacttctgc ggtggctccc tcatcagcga acagtgggtg 180
gtgtcagcag gtcactgcta caagtcccgc atccaggtga gactgggaga gcacaacatc 240
gaagtcctgg aggggaatga acagttcatc aatgcagcca agatcatccg ccaccccaaa 300
tacaacagcc ggactctgga caatgacatc ctgctgatca agctctcctc acctgccgtc 360
atcaattccc gcgtgtccgc catctctctg cccactgccc ctccagctgc tggcaccgag 420
tccctcatct ccggctgggg caacactctg agttctggtg ccgactaccc agacgagctg 480
cagtgcctgg atgctcctgt gctgagccag gctgagtgtg aagcctccta ccctggaaag 540
attaccaaca acatgttctg tgtgggcttc ctcgagggag gcaaggattc ctgccagggt 600
gattctggtg gccctgtggt ctccaatgga gagctccaag gaattgtctc ctggggctat 660
ggctgtgccc agaagaacag gcctggagtc tacaccaagg tctacaacta tgtggactgg 720
attaaggaca ccatagctgc caacagctaa agcccctggt ccctctgcag tctctatacc 780
aataaagtga ccctgctctc ac 802
<210>237
<211>247
<212>PRT
<213>Homo sapiens
<220>
<223>anionic trypsin (II)
<400>237
Met Asn Leu Leu Leu Ile Leu Thr Phe Val Ala Ala Ala Val Ala Ala
1 5 10 15
Pro Phe Asp Asp Asp Asp Lys Ile Val Gly Gly Tyr Ile Cys Glu Glu
20 25 30
Asn Ser Val Pro Tyr Gln Val Ser Leu Asn Ser Gly Tyr His Phe Cys
35 40 45
Gly Gly Ser Leu Ile Ser Glu Gln Trp Val Val Ser Ala Gly His Cys
50 55 60
Tyr Lys Ser Arg Ile Gln Val Arg Leu Gly Glu His Asn Ile Glu Val
65 70 75 80
Leu Glu Gly Asn Glu Gln Phe Ile Asn Ala Ala Lys Ile Ile Arg His
85 90 95
Pro Lys Tyr Asn Ser Arg Thr Leu Asp Asn Asp Ile Leu Leu Ile Lys
100 105 110
Leu Ser Ser Pro Ala Val Ile Asn Ser Arg Val Ser Ala Ile Ser Leu
115 120 125
Pro Thr Ala Pro Pro Ala Ala Gly Thr Glu Ser Leu Ile Ser Gly Trp
130 135 140
Gly Asn Thr Leu Ser Ser Gly Ala Asp Tyr Pro Asp Glu Leu Gln Cys
145 150 155 160
Leu Asp Ala Pro Val Leu Ser Gln Ala Glu Cys Glu Ala Ser Tyr Pro
165 170 175
Gly Lys Ile Thr Asn Asn Met Phe Cys Val Gly Phe Leu Glu Gly Gly
180 185 190
Lys Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Val Val Ser Asn Gly
195 200 205
Glu Leu Gln Gly Ile Val Ser Trp Gly Tyr Gly CysAla Gln Lys Asn
210 215 220
Arg Pro Gly Val Tyr Thr Lys Val Tyr Asn Tyr Val Asp Trp Ile Lys
225 230 235 240
Asp Thr Ile Ala Ala Asn Ser
245
<210>238
<211>1475
<212>DNA
<213>Homo sapiens
<220>
<223>LOC144757 peptidase
<400>238
acctgccgcc atcttgctca ccagcctcca aaatgcggct ggggctcctg agcgtggcgc 60
tgttgtttgt ggggagctct cacttatact cagaccacta ctcgccctct ggaaggcaca 120
ggctcggccc ctcgccggaa ccggcggcta gttcccagca ggctgaggcc gtccgcaaga 180
ggctccggcg gcggagggag ggaggggcgc atgcaaagga ttgtggaaca gcaccgctta 240
aggatgtgtt gcaagggtct cggattatag ggggcaccga agcacaagct ggcgcatggc 300
cgtgggtggt gagcctgcag attaaatatg gccgtgttct tgttcatgta tgtgggggaa 360
ccctagtgag agagaggtgg gtcctcacag ctgcccactg cactaaagac gctagcgatc 420
ctttaatgtg gacagctgtg attggaacta ataatataca tggacgctat cctcatacca 480
agaagataaa aattaaagca atcattattc atccaaactt cattttggaa tcttatgtaa 540
atgatattgc actttttcac ttaaaaaaag cagtgaggta taatgactat attcagccta 600
tttgcctacc ttttgatgtt ttccaaatcc tggacggaaa cacaaagtgt tttataagtg 660
gctggggaag aacaaaagaa gaaggtaacg ctacaaatat tttacaagat gcagaagtgc 720
attatatttc tcgagagatg tgtaattctg agaggagtta tgggggaata attcctaaca 780
cttcattttg tgcaggtgat gaagatggag cttttgatac ttgcaggggt gacagtgggg 840
gaccattaat gtgctactta ccagaatata aaagattttt tgtaatggga attaccagtt 900
acggacatgg ctgtggtcga agaggttttc ctggtgtcta tattgggcca tccttctacc 960
aaaagtggct gacagagcat ttcttccatg caagcactca aggcatactt actataaata 1020
ttttacgtgg ccagatcctc atagctttat gttttgtcat cttactagca acaacataaa 1080
gaaattctga aggctttcat atctttattt tgcattgtgt ccctttctat gttctatata 1140
atgaacatca tttattcttc tagcaattaa ttgcctacat tagagatttc atgtgaacat 1200
tttatgggct ataagtattg tgacagatat acaattgtaa ttttggcact gaatcacatg 1260
tctccttgaa atatcttgat tattttataa tcataattct gtatctggaa tactcataga 1320
gtttgtacaa aatattcagt taaacatata tgtgtataaa tgccaaataa tagtttataa 1380
ttaaaatgaa agctgtcatt tggttaaatt aataaaaatt ctttcttaga ttttattcta 1440
aaaaaaaaaa aaaaaaaaga aaaaaaaaaa aaaaa 1475
<210>239
<211>348
<212>PRT
<213>Homo sapiens
<220>
<223>LOC144757 peptidase
<400>239
Met Arg Leu Gly Leu Leu Ser Val Ala Leu Leu Phe Val Gly Ser Ser
1 5 10 15
His Leu Tyr Ser Asp His Tyr Ser Pro Ser Gly Arg His Arg Leu Gly
20 25 30
Pro Ser Pro Glu Pro Ala Ala Ser Ser Gln Gln Ala Glu Ala Val Arg
35 40 45
Lys Arg Leu Arg Arg Arg Arg Glu Gly Gly Ala His Ala Lys Asp Cys
50 55 60
Gly Thr Ala Pro Leu Lys Asp Val Leu Gln Gly Ser Arg Ile Ile Gly
65 70 75 80
Gly Thr Glu Ala Gln Ala Gly Ala Trp Pro Trp Val Val Ser Leu Gln
85 90 95
Ile Lys Tyr Gly Arg Val Leu Val His Val Cys Gly Gly Thr Leu Val
100 105 110
Arg Glu Arg Trp Val Leu Thr Ala Ala His Cys Thr Lys Asp Ala Ser
115 120 125
Asp Pro Leu Met Trp Thr Ala Val Ile Gly Thr Asn Asn Ile His Gly
130 135 140
Arg Tyr Pro His Thr Lys Lys Ile Lys Ile Lys Ala Ile Ile Ile His
145 150 155 160
Pro Asn Phe Ile Leu Glu Ser Tyr Val Asn Asp Ile Ala Leu Phe His
165 170 175
Leu Lys Lys Ala Val Arg Tyr Asn Asp Tyr Ile Gln Pro Ile Cys Leu
180 185 190
Pro Phe Asp Val Phe Gln Ile Leu Asp Gly Asn Thr Lys Cys Phe Ile
195 200 205
Ser Gly Trp Gly Arg Thr Lys Glu Glu Gly Asn Ala Thr Asn Ile Leu
210 215 220
Gln Asp Ala Glu Val His Tyr Ile Ser Arg Glu Met Cys Asn Ser Glu
225 230 235 240
Arg Ser Tyr Gly Gly Ile Ile Pro Asn Thr Ser Phe Cys Ala Gly Asp
245 250 255
Glu Asp Gly Ala Phe Asp Thr Cys Arg Gly Asp Ser Gly Gly Pro Leu
260 265 270
Met Cys Tyr Leu Pro Glu Tyr Lys Arg Phe Phe Val Met Gly Ile Thr
275 280 285
Ser Tyr Gly His Gly Cys Gly Arg Arg Gly Phe Pro Gly Val Tyr Ile
290 295 300
Gly Pro Ser Phe Tyr Gln Lys Trp Leu Thr Glu His Phe Phe His Ala
305 310 315 320
Ser Thr Gln Gly Ile Leu Thr Ile Asn Ile Leu Arg Gly Gln Ile Leu
325 330 335
Ile Ala Leu Cys Phe Val Ile Leu Leu Ala Thr Thr
340 345
<210>240
<211>1164
<212>DNA
<213>Homo sapiens
<220>
<223>Mername-AA169 peptidase
<400>240
atgattgccg ttctcattgt gttgtccctg acagtggtgg cagtgaccat aggtctcctg 60
gttcacttcc tagtatttga ccaaaaaaag gagtactatc atggctcctt taaaatttta 120
gatccacaaa tcaataacaa tttcggacaa agcaacacat atcaacttaa ggacttacga 180
gagacgaccg aaaatttggt ggatgagata tttatagatt cagcctggaa gaaaaattat 240
atcaagaacc aagtagtcag actgactcca gaggaagatg gtgtgaaagt agatgtcatt 300
atggtgttcc agttcccctc tactgaacaa agggcagtaa gagagaagaa aatccaaagc 360
atcttaaatc agaagataag gaatttaaga gccttgccaa taaatgcctc atcagttcaa 420
gttaatggtt gtggtaaacg agttgttcca ttaaacgtca acagaatagc atctggagtc 480
attgcaccca aggcggcctg gccttggcaa gcttcccttc agtatgataa catccatcag 540
tgtggggcca ccttgattag taacacatgg cttgtcactg cagcacactg cttccagaag 600
tataaaaatc cacatcaatg gactgttagt tttggaacaa aaatcaaccc tcccttaatg 660
aaaagaaatg tcagaagatt tattatccat gagaagtacc gctctgcagc aagagagtac 720
gacattgctg ttgtgcaggt ctcttccaga gtcacctttt cggatgacat acgccagatt 780
tgtttgccag aagcctctgc atccttccaa ccaaatttga ctgtccacat cacaggattt 840
ggagcacttt actatggtgg ggaatcccaa aatgatctcc gagaagccag agtgaaaatc 900
ataagtgatg atgtctgcaa gcaaccacag gtgtatggca atgatataaa acctggaatg 960
ttctgtgccg gatatatgga aggaatttat gatgcctgca ggggtgattc tgggggacct 1020
ttagtcacaa gggatctgaa agatacgtgg tatctcattg gaattgtaag ctggggagat 1080
aactgtggtc aaaaggacaa gcctggagtc tacacacaag tgacttatta ccgaaactgg 1140
attgcttcaa aaacaggcat ctaa 1164
<210>241
<211>387
<212>PRT
<213>Homo sapiens
<220>
<223>Mername-AA169 peptidase
<400>241
Met Ile Ala Val Leu Ile Val Leu Ser Leu Thr Val Val Ala Val Thr
1 5 10 15
Ile Gly Leu Leu Val His Phe Leu Val Phe Asp Gln Lys Lys Glu Tyr
20 25 30
Tyr His Gly Ser Phe Lys Ile Leu Asp Pro Gln Ile Asn Asn Asn Phe
35 40 45
Gly Gln Ser Asn Thr Tyr Gln Leu Lys Asp Leu Arg Glu Thr Thr Glu
50 55 60
Asn Leu Val Asp Glu Ile Phe Ile Asp Ser Ala Trp Lys Lys Asn Tyr
65 70 75 80
Ile Lys Asn Gln Val Val Arg Leu Thr Pro Glu Glu Asp Gly Val Lys
85 90 95
Val Asp Val Ile Met Val Phe Gln Phe Pro Ser Thr Glu Gln Arg Ala
100 105 110
Val Arg Glu Lys Lys Ile Gln Ser Ile Leu Asn Gln Lys Ile Arg Asn
115 120 125
Leu Arg Ala Leu Pro Ile Asn Ala Ser Ser Val Gln Val Asn Gly Cys
130 135 140
Gly Lys Arg Val Val Pro Leu Asn Val Asn Arg Ile Ala Ser Gly Val
145 150 155 160
Ile Ala Pro Lys Ala Ala Trp Pro Trp Gln Ala Ser Leu Gln Tyr Asp
165 170 175
Asn Ile His Gln Cys Gly Ala Thr Leu Ile Ser Asn Thr Trp Leu Val
180 185 190
Thr Ala Ala His Cys Phe Gln Lys Tyr Lys Asn Pro His Gln Trp Thr
195 200 205
Val Ser Phe Gly Thr Lys Ile Asn Pro Pro Leu Met Lys Arg Asn Val
210 215 220
Arg Arg Phe Ile Ile His Glu Lys Tyr Arg Ser Ala Ala Arg Glu Tyr
225 230 235 240
Asp Ile Ala Val Val Gln Val Ser Ser Arg Val Thr Phe Ser Asp Asp
245 250 255
Ile Arg Gln Ile Cys Leu Pro Glu Ala Ser Ala Ser Phe Gln Pro Asn
260 265 270
Leu Thr Val His Ile Thr Gly Phe Gly Ala Leu Tyr Tyr Gly Gly Glu
275 280 285
Ser Gln Asn Asp Leu Arg Glu Ala Arg ValLys Ile Ile Ser Asp Asp
290 295 300
Val Cys Lys Gln Pro Gln Val Tyr Gly Asn Asp Ile Lys Pro Gly Met
305 310 315 320
Phe Cys Ala Gly Tyr Met Glu Gly Ile Tyr Asp Ala Cys Arg Gly Asp
325 330 335
Ser Gly Gly Pro Leu Val Thr Arg Asp Leu Lys Asp Thr Trp Tyr Leu
340 345 350
Ile Gly Ile Val Ser Trp Gly Asp Asn Cys Gly Gln Lys Asp Lys Pro
355 360 365
Gly Val Tyr Thr Gln Val Thr Tyr Tyr Arg Asn Trp Ile Ala Ser Lys
370 375 380
Thr Gly Ile
385
<210>242
<211>431
<212>PRT
<213>Rattus norvegicus
<220>
<223>Mername-AA171 peptidase
<400>242
Met Ala Arg Gly Gln Pro Arg Arg Ser Glu Glu Pro Trp Thr Ala Leu
1 5 10 15
Arg Asn Arg Thr Glu Cys Lys Thr Lys Ile Lys Leu Thr Lys Cys Gly
20 25 30
Lys Ile Thr Leu Gly Ile Leu Thr Ala Val Ile Ala Ala Val Val Ile
35 40 45
Gly Leu Ile Ala Tyr Phe Ala Ala Cys Gly Lys Thr Pro Phe Tyr Tyr
50 55 60
His Val Ser Phe Lys Val Asn Asn Ile Asp Tyr Asp Ser Lys Phe Ala
65 70 75 80
Lys Pro Tyr Ser Gln Glu Tyr Met Asp Leu Asn Lys Arg Ile Val Ser
85 90 95
Leu Met Asn Glu Thr Phe His Glu Ser Lys Leu Arg Lys Gln Tyr Val
100 105 110
Lys Ala His Thr Val Gln Val Ser Lys Ala Lys Gly Lys Val Val Ile
115 120 125
His Gly Val Leu Lys Phe Lys Ser Cys Tyr Lys Asn Asn Val Glu Lys
130 135 140
Phe Trp Asp Ser Val Glu Thr Ile Leu Tyr Gln Lys Leu Lys Ser Gln
145 150 155 160
Thr Arg Leu Leu Ile Asp Ser Ser Ser Phe Lys Phe Ser Asp Ile Ala
165 170 175
Met Pro Ile Ala Glu Asp Leu Leu Asn Thr Cys Cys Gly Arg Arg Thr
180 185 190
Ile Thr Pro Gly Gly His Lys Val Ala Gly Gly Gln Asp Ala Glu Glu
195 200 205
Gly Glu Trp Pro Trp Gln Ala Ser Leu Gln Gln Asn Asn Val His Arg
210 215 220
Cys Gly Ala Thr Leu Ile Ser Asn Ser Trp Leu Ile Thr Ala Ala His
225 230 235 240
Cys Phe Val Arg Ser Ala Asn Pro Lys Asp Trp Lys Val Ser Phe Gly
245 250 255
Phe Leu Leu Ser Lys Pro Gln Ala Gln Arg Ala Val Lys Ser Ile Val
260 265 270
Ile His Glu Asn Tyr Ser Tyr Pro Ala His Asn Asn Asp Ile Ala Val
275 280 285
Val Arg Leu Ser Ser Pro Val Leu Tyr Glu Asn Asn Ile Arg Arg Ala
290 295 300
Cys Leu Pro Glu Ala Thr Gln Lys Phe Pro Pro Asn Ser Asp Val Val
305 310 315 320
ValThr Gly Trp Gly Thr Leu Lys Ser Asp Gly Asp Ser Pro Asn Ile
325 330 335
Leu Gln Lys Gly Arg Val Lys Ile Ile Asp Asn Lys Thr Cys Asn Ser
340 345 350
Gly Lys Ala Tyr Gly Gly Val Ile Thr Pro Gly Met Leu Cys Ala Gly
355 360 365
Phe Leu Glu Gly Arg Val Asp Ala Cys Gln Gly Asp Ser Gly Gly Pro
370 375 380
Leu Val Ser Glu Asp Ser Lys Gly Ile Trp Phe Leu Ala Gly Ile Val
385 390 395 400
Ser Trp Gly Asp Glu Cys Ala Leu Pro Asn Lys Pro Gly Val Tyr Thr
405 410 415
Arg Val Thr His Tyr Arg Asp Trp Ile Ser Ser Lys Thr Gly Leu
420 425 430
<210>243
<211>247
<212>PRT
<213>Homo sapiens
<220>
<223>Mername-AA174 peptidase
<400>243
Met Asn Pro Leu Leu Ile Leu Ala Phe Val Gly Ala Ala Val Ala Val
1 5 10 15
Pro Phe Asp Asp Asp Asp Lys Ile Val Gly Gly Tyr Thr Cys Glu Glu
20 25 30
Asn Ser Val Pro Tyr Gln ValSer Leu Asn Ser Gly Ser His Phe Cys
35 40 45
Gly Gly Ser Leu Ile Ser Glu Gln Trp Val Val Ser Ala Gly His Cys
50 55 60
Tyr Lys Pro His Ile Gln ValArg Leu Gly Glu His Asn Ile Glu Val
65 70 75 80
Leu Glu Gly Asn Glu Gln Phe Ile Asn Ala Ala Lys Ile Ile Arg His
85 90 95
Pro Lys Tyr Asn Arg Ile Thr Leu Asn Asn Asp Ile Met Leu Ile Lys
100 105 110
Leu Ser Thr Pro Ala Val Ile Asn Ala His Val Ser Thr Ile Ser Leu
115 120 125
Pro Thr Ala Pro Pro Ala Ala Gly Thr Glu Cys Leu Ile Ser Gly Trp
130 135 140
Gly Asn Thr Leu Ser Ser Gly Ala Asp Tyr Pro Asp Glu Leu Gln Cys
145 150 155 160
Leu Asp Ala Pro Val Leu Thr Gln Ala Lys Cys Lys Ala Ser Tyr Pro
165 170 175
Leu Lys Ile Thr Ser Lys Met Phe Cys Val Gly Phe Leu Glu Gly Gly
180 185 190
Lys Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro ValVal Cys Asn Gly
195 200 205
Gln Leu Gln Gly Ile ValSer Trp Gly Tyr Gly Cys Ala Gln Lys Arg
210 215 220
Arg Pro Gly Val Tyr Thr Lys Val Tyr Asn Tyr Val Asp Trp Ile Lys
225 230 235 240
Asp Thr Ile Ala Ala Asn Ser
245
<210>244
<211>1099
<212>DNA
<213>Homo sapiens
<220>
<223>Mername-AA175 peptidase
<400>244
gcacagccca gggccatgct cctgttctca gtgttgctgc tcctgtccct ggtcacggga 60
actcagctcg gtccacggac tcctctccca gaggctggag tggctatcct aggcagggct 120
aggggagccc accgccctca gccccgtcat ccccccagcc cagtcagtga atgtggtgac 180
agatctattt tcgagggaag aactcggtat tccagaatca caggggggat ggaggcggag 240
gtgggtgagt ttccgtggca ggtgagtatt caggcaagaa gtgaaccttt ctgtggcggc 300
tccatcctca acaagtggtg gattctcact gcggctcact gcttatattc cgaggagctg 360
tttccagaag aactgagtgt cgtgctgggg accaacgact taactagccc atccatggaa 420
ataaaggagg tcgccagcat cattcttcac aaagacttta agagagccaa catggacaat 480
gacattgcct tgctgctgct ggcttcgccc atcaagctcg atgacctgaa ggtgcccatc 540
tgcctcccca cgcagcccgg ccctgccaca tggcgcgaat gctgggtggc aggttggggc 600
cagaccaatg ctgctgacaa aaactctgtg aaaacggatc tgatgaaagt gccaatggtc 660
atcatggact gggaggagtg ttcaaagatg tttccaaaac ttaccaaaaa tatgctgtgt 720
gccggataca agaatgagag ctatgatgcc tgcaagggtg acagtggggg gcctctggtc 780
tgcaccccag agcctggtga gaagtggtac caggtgggca tcatcagctg gggaaagagc 840
tgtggagaga agaacacccc agggatatac acctcgttgg tgaactacaa cctctggatc 900
gagaaagtga cccagctaga gggcaggccc ttcaatgcag agaaaaggag gacttctgtc 960
aaacagaaac ctatgggctc cccagtctcg ggagtcccag agccaggcag ccccagatcc 1020
tggctcctgc tctgtcccct gtcccatgtg ttgttcagag ctattttgta ctgataataa 1080
aatagaggct attctttca 1099
<210>245
<211>352
<212>PRT
<213>Homo sapiens
<220>
<223>Mername-AA175 peptidase
<400>245
Met Leu Leu Phe Ser Val Leu Leu Leu Leu Ser Leu Val Thr Gly Thr
1 5 10 15
Gln Leu Gly Pro Arg Thr Pro Leu Pro Glu Ala Gly Val Ala Ile Leu
20 25 30
Gly Arg Ala Arg Gly Ala His Arg Pro Gln Pro Arg His Pro Pro Ser
35 40 45
Pro Val Ser Glu Cys Gly Asp Arg Ser Ile Phe Glu Gly Arg Thr Arg
50 55 60
Tyr Ser Arg Ile Thr Gly Gly Met Glu Ala Glu Val Gly Glu Phe Pro
65 70 75 80
Trp Gln Val Ser Ile Gln Ala Arg Ser Glu Pro Phe Cys Gly Gly Ser
85 90 95
Ile Leu Asn Lys Trp Trp Ile Leu Thr Ala Ala His Cys Leu Tyr Ser
100 105 110
Glu Glu Leu Phe Pro Glu Glu Leu Ser Val Val Leu Gly Thr Asn Asp
115 120 125
Leu Thr Ser Pro Ser Met Glu Ile Lys Glu Val Ala Ser Ile Ile Leu
130 135 140
His Lys Asp Phe Lys Arg Ala Asn Met Asp Asn Asp Ile Ala Leu Leu
145 150 155 160
Leu Leu Ala Ser Pro Ile Lys Leu Asp Asp Leu Lys Val Pro Ile Cys
165 170 175
Leu Pro Thr Gln Pro Gly Pro Ala Thr Trp Arg Glu Cys Trp Val Ala
180 185 190
Gly Trp Gly Gln Thr Asn Ala Ala Asp Lys Asn Ser Val Lys Thr Asp
195 200 205
Leu Met Lys Val Pro Met Val Ile Met Asp Trp Glu Glu Cys Ser Lys
210 215 220
Met Phe Pro Lys Leu Thr Lys Asn Met Leu Cys Ala Gly Tyr Lys Asn
225 230 235 240
Glu Ser Tyr Asp Ala Cys Lys Gly Asp Ser Gly Gly Pro Leu Val Cys
245 250 255
Thr Pro Glu Pro Gly Glu Lys Trp Tyr Gln Val Gly Ile Ile Ser Trp
260 265 270
Gly Lys Ser Cys Gly Glu Lys Asn Thr Pro Gly Ile Tyr Thr Ser Leu
275 280 285
Val Asn Tyr Asn Leu Trp Ile Glu Lys Val Thr Gln Leu Glu Gly Arg
290 295 300
Pro Phe Asn Ala Glu Lys Arg Arg Thr Ser Val Lys Gln Lys Pro Met
305 310 315 320
Gly Ser Pro Val Ser Gly Val Pro Glu Pro Gly Ser Pro Arg Ser Trp
325 330 335
Leu Leu Leu Cys Pro Leu Ser His Val Leu Phe Arg Ala Ile Leu Tyr
340 345 350
<210>246
<211>1927
<212>DNA
<213>Homo sapiens
<220>
<223>stratum corneum chymotryptic enzyme transcript variant 1
<400>246
tgccagccca agtcggaact tggatcacat cagatcctct cgagctccag caggagaggc 60
ccttcctcgc ctggcagccc ctgagcggct cagcagggca ccatggcaag atcccttctc 120
ctgcccctgc agatcttact gctatcctta gccttggaaa ctgcaggaga agaagcccag 180
ggtgacaaga ttattgatgg cgccccatgt gcaagaggct cccacccatg gcaggtggcc 240
ctgctcagtg gcaatcagct ccactgcgga ggcgtcctgg tcaatgagcg ctgggtgctc 300
actgccgccc actgcaagat gaatgagtac accgtgcacc tgggcagtga tacgctgggc 360
gacaggagag ctcagaggat caaggcctcg aagtcattcc gccaccccgg ctactccaca 420
cagacccatg ttaatgacct catgctcgtg aagctcaata gccaggccag gctgtcatcc 480
atggtgaaga aagtcaggct gccctcccgc tgcgaacccc ctggaaccac ctgtactgtc 540
tccggctggg gcactaccac gagcccagat gtgacctttc cctctgacct catgtgcgtg 600
gatgtcaagc tcatctcccc ccaggactgc acgaaggttt acaaggactt actggaaaat 660
tccatgctgt gcgctggcat ccccgactcc aagaaaaacg cctgcaatgg tgactcaggg 720
ggaccgttgg tgtgcagagg taccctgcaa ggtctggtgt cctggggaac tttcccttgc 780
ggccaaccca atgacccagg agtctacact caagtgtgca agttcaccaa gtggataaat 840
gacaccatga aaaagcatcg ctaacgccac actgagttaa ttaactgtgt gcttccaaca 900
gaaaatgcac aggagtgagg acgccgatga cctatgaagt caaatttgac tttacctttc 960
ctcaaagata tatttaaacc aacctcatgc cctgttgata aaccaatcaa attggtaaag 1020
acctaaaacc aaaacaaata aagaaacaca aaaccctcag tgctggagaa gagtcagtga 1080
gaccagcact ctcaaacact ggaactggac gttcgtacag tctttacgga agacacttgg 1140
tcaacgtaca ccgagaccct tattcaccac ctttgaccca gtaactctaa tcttaggaag 1200
aacctactga aacaaaaaaa atccaaaatg tagaacaaga cttgaattta ccatgatatt 1260
atttatcaca gaaatgaagt gaaaccatca aacatgttcc aaaagtacca gatggcttaa 1320
ataatagtct ggcttggcac aacgatgttt tttttctttg agacagagtc tctgttgctt 1380
gggctgcaat gcagtgatgc aatcttggct cactgcaacc tccgcctcct gggttcaagt 1440
gattctcgtg cttcagcctc ccaagtacct gggactacag gtgtgcacca ccacaccagg 1500
ctaatttttt gtgtattttt actagagaca gggtttcacc atgttggcca gcgtggtctt 1560
gaacgcctga cctcagatga tccacccacc ttggcctccc aaagtgctgg gattacaggc 1620
atgagccacc acggccagcc cacaatgata ttacaaacct attaaaaatg atacttagac 1680
agaattgtca gtattattca agaacattta ggctatagga tgttaaatga caaaaggaag 1740
gacaaaaata tatatgtatg tgaccctacc cataaaaaat gaaatattca cagaatcaga 1800
tctgaaaaca catgtcccag actgcatact ggggtcgtca tgaggtgtct ccttccttct 1860
gtgtactttt ccttgaatgt gcacttttat aacatgaaaa ataaaggtgg ggaaaaaagt 1920
ctgaaga 1927
<210>247
<211>253
<212>PRT
<213>Homo sapiens
<220>
<223>stratum corneum chymotryptic enzyme transcript variant 1
<400>247
Met Ala Arg Ser Leu Leu Leu Pro Leu Gln Ile Leu Leu Leu Ser Leu
1 5 10 15
Ala Leu Glu Thr Ala Gly Glu Glu Ala Gln Gly Asp Lys Ile Ile Asp
20 25 30
Gly Ala Pro Cys Ala Arg Gly Ser His Pro Trp Gln Val Ala Leu Leu
35 40 45
Ser Gly Asn Gln Leu His Cys Gly Gly Val Leu Val Asn Glu Arg Trp
50 55 60
Val Leu Thr Ala Ala His Cys Lys Met Asn Glu Tyr Thr Val His Leu
65 70 75 80
Gly Ser Asp Thr Leu Gly Asp Arg Arg Ala Gln Arg Ile Lys Ala Ser
85 90 95
Lys Ser Phe Arg His Pro Gly Tyr Ser Thr Gln Thr His Val Asn Asp
100 105 110
Leu Met Leu Val Lys Leu Asn Ser Gln Ala Arg Leu Ser Ser Met Val
115 120 125
Lys Lys Val Arg Leu Pro Ser Arg Cys Glu Pro Pro Gly Thr Thr Cys
130 135 140
Thr Val Ser Gly Trp Gly Thr Thr Thr Ser Pro Asp Val Thr Phe Pro
145 150 155 160
Ser Asp Leu Met Cys Val Asp Val Lys Leu Ile Ser Pro Gln Asp Cys
165 170 175
Thr Lys Val Tyr Lys Asp Leu Leu Glu Asn Ser Met Leu Cys Ala Gly
180 185 190
Ile Pro Asp Ser Lys Lys Asn Ala Cys Asn Gly Asp Ser Gly Gly Pro
195 200 205
Leu Val Cys Arg Gly Thr Leu Gln Gly Leu Val Ser Trp Gly Thr Phe
210 215 220
Pro Cys Gly Gln Pro Asn Asp Pro Gly Val Tyr Thr Gln Val Cys Lys
225 230 235 240
Phe Thr Lys Trp Ile Asn Asp Thr Met Lys Lys His Arg
245 250
<210>248
<211>1927
<212>DNA
<213>Homo sapiens
<220>
<223>stratum corneum chymotryptic enzyme transcript variant 2
<400>248
ggagagggtg atgctggctc caagcctgac tctgctctcg agagctccag caggagaggc 60
ccttcctcgc ctggcagccc ctgagcggct cagcagggca ccatggcaag atcccttctc 120
ctgcccctgc agatcttact gctatcctta gccttggaaa ctgcaggaga agaagcccag 180
ggtgacaaga ttattgatgg cgccccatgt gcaagaggct cccacccatg gcaggtggcc 240
ctgctcagtg gcaatcagct ccactgcgga ggcgtcctgg tcaatgagcg ctgggtgctc 300
actgccgccc actgcaagat gaatgagtac accgtgcacc tgggcagtga tacgctgggc 360
gacaggagag ctcagaggat caaggcctcg aagtcattcc gccaccccgg ctactccaca 420
cagacccatg ttaatgacct catgctcgtg aagctcaata gccaggccag gctgtcatcc 480
atggtgaaga aagtcaggct gccctcccgc tgcgaacccc ctggaaccac ctgtactgtc 540
tccggctggg gcactaccac gagcccagat gtgacctttc cctctgacct catgtgcgtg 600
gatgtcaagc tcatctcccc ccaggactgc acgaaggttt acaaggactt actggaaaat 660
tccatgctgt gcgctggcat ccccgactcc aagaaaaacg cctgcaatgg tgactcaggg 720
ggaccgttgg tgtgcagagg taccctgcaa ggtctggtgt cctggggaac tttcccttgc 780
ggccaaccca atgacccagg agtctacact caagtgtgca agttcaccaa gtggataaat 840
gacaccatga aaaagcatcg ctaacgccac actgagttaa ttaactgtgt gcttccaaca 900
gaaaatgcac aggagtgagg acgccgatga cctatgaagt caaatttgac tttacctttc 960
ctcaaagata tatttaaacc aacctcatgc cctgttgata aaccaatcaa attggtaaag 1020
acctaaaacc aaaacaaata aagaaacaca aaaccctcag tgctggagaa gagtcagtga 1080
gaccagcact ctcaaacact ggaactggac gttcgtacag tctttacgga agacacttgg 1140
tcaacgtaca ccgagaccct tattcaccac ctttgaccca gtaactctaa tcttaggaag 1200
aacctactga aacaaaaaaa atccaaaatg tagaacaaga cttgaattta ccatgatatt 1260
atttatcaca gaaatgaagt gaaaccatca aacatgttcc aaaagtacca gatggcttaa 1320
ataatagtct ggcttggcac aacgatgttt tttttctttg agacagagtc tctgttgctt 1380
gggctgcaat gcagtgatgc aatcttggct cactgcaacc tccgcctcct gggttcaagt 1440
gattctcgtg cttcagcctc ccaagtacct gggactacag gtgtgcacca ccacaccagg 1500
ctaatttttt gtgtattttt actagagaca gggtttcacc atgttggcca gcgtggtctt 1560
gaacgcctga cctcagatga tccacccacc ttggcctccc aaagtgctgg gattacaggc 1620
atgagccacc acggccagcc cacaatgata ttacaaacct attaaaaatg atacttagac 1680
agaattgtca gtattattca agaacattta ggctatagga tgttaaatga caaaaggaag 1740
gacaaaaata tatatgtatg tgaccctacc cataaaaaat gaaatattca cagaatcaga 1800
tctgaaaaca catgtcccag actgcatact ggggtcgtca tgaggtgtct ccttccttct 1860
gtgtactttt ccttgaatgt gcacttttat aacatgaaaa ataaaggtgg ggaaaaaagt 1920
ctgaaga 1927
<210>249
<211>253
<212>PRT
<213>Homo sapiens
<220>
<223>stratum corneum chymotryptic enzyme
<400>249
Met Ala Arg Ser Leu Leu Leu Pro Leu Gln Ile Leu Leu Leu Ser Leu
1 5 10 15
Ala Leu Glu Thr Ala Gly Glu Glu Ala Gln Gly Asp Lys Ile Ile Asp
20 25 30
Gly Ala Pro Cys Ala Arg Gly Ser His Pro Trp Gln Val Ala Leu Leu
35 40 45
Ser Gly Asn Gln Leu His Cys Gly Gly Val Leu Val Asn Glu Arg Trp
50 55 60
Val Leu Thr Ala Ala His Cys Lys Met Asn Glu Tyr Thr Val His Leu
65 70 75 80
Gly Ser Asp Thr Leu Gly Asp Arg Arg Ala Gln Arg Ile Lys Ala Ser
85 90 95
Lys Ser Phe Arg His Pro Gly Tyr Ser Thr Gln Thr His Val Asn Asp
100 105 110
Leu Met Leu Val Lys Leu Asn Ser Gln Ala Arg Leu Ser Ser Met Val
115 120 125
Lys Lys Val Arg Leu Pro Ser Arg Cys Glu Pro Pro Gly Thr Thr Cys
130 135 140
Thr Val Ser Gly Trp Gly Thr Thr Thr Ser Pro Asp Val Thr Phe Pro
145 150 155 160
Ser Asp Leu Met Cys Val Asp Val Lys Leu Ile Ser Pro Gln Asp Cys
165 170 175
Thr Lys Val Tyr Lys Asp Leu Leu Glu Asn Ser Met Leu Cys Ala Gly
180 185 190
Ile Pro Asp Ser Lys Lys Asn Ala Cys Asn Gly Asp Ser Gly Gly Pro
195 200 205
Leu Val Cys Arg Gly Thr Leu Gln Gly Leu Val Ser Trp Gly Thr Phe
210 215 220
Pro Cys Gly Gln Pro Asn Asp Pro Gly Val Tyr Thr Gln Val Cys Lys
225 230 235 240
Phe Thr Lys Trp Ile Asn Asp Thr Met Lys Lys His Arg
245 250
<210>250
<211>2800
<212>DNA
<213>Homo sapiens
<220>
<223>trypsin-like enzyme，respiratory
<400>250
atttgagtgg gaatctcaaa gcagttgagt aggcagaaaa aagaacctct tcattaagga 60
ttaaaatgta taggccagca cgtgtaactt cgacttcaag atttctgaat ccatatgtag 120
tatgtttcat tgtcgtcgca ggggtagtga tcctggcagt caccatagct ctacttgttt 180
actttttagc ttttgatcaa aaatcttact tttataggag cagttttcaa ctcctaaatg 240
ttgaatataa tagtcagtta aattcaccag ctacacagga atacaggact ttgagtggaa 300
gaattgaatc tctgattact aaaacattca aagaatcaaa tttaagaaat cagttcatca 360
gagctcatgt tgccaaactg aggcaagatg gtagtggtgt gagagcggat gttgtcatga 420
aatttcaatt cactagaaat aacaatggag catcaatgaa aagcagaatt gagtctgttt 480
tacgacaaat gctgaataac tctggaaacc tggaaataaa cccttcaact gagataacat 540
cacttactga ccaggctgca gcaaattggc ttattaatga atgtggggcc ggtccagacc 600
taataacatt gtctgagcag agaatccttg gaggcactga ggctgaggag ggaagctggc 660
cgtggcaagt cagtctgcgg ctcaataatg cccaccactg tggaggcagc ctgatcaata 720
acatgtggat cctgacagca gctcactgct tcagaagcaa ctctaatcct cgtgactgga 780
ttgccacgtc tggtatttcc acaacatttc ctaaactaag aatgagagta agaaatattt 840
taattcataa caattataaa tctgcaactc atgaaaatga cattgcactt gtgagacttg 900
agaacagtgt cacctttacc aaagatatcc atagtgtgtg tctcccagct gctacccaga 960
atattccacc tggctctact gcttatgtaa caggatgggg cgctcaagaa tatgctggcc 1020
acacagttcc agagctaagg caaggacagg tcagaataat aagtaatgat gtatgtaatg 1080
caccacatag ttataatgga gccatcttgt ctggaatgct gtgtgctgga gtacctcaag 1140
gtggagtgga cgcatgtcag ggtgactctg gtggcccact agtacaagaa gactcacggc 1200
ggctttggtt tattgtgggg atagtaagct ggggagatca gtgtggcctg ccggataagc 1260
caggagtgta tactcgagtg acagcctacc ttgactggat taggcaacaa actgggatct 1320
agtgcaacaa gtgcatccct gttgcaaagt ctgtatgcag gtgtgcctgt cttaaattcc 1380
aaagctttac atttcaactg aaaaagaaac tagaaatgtc ctaatttaac atcttgttac 1440
ataaatatgg tttaacaaac actgtttaac ctttctttat tattaaaggt tttctatttt 1500
ctccagagaa ctatatgaat gttgcatagt actgtggctg tgtaacagaa gaaacacact 1560
aaactaatta caaagttaac aatttcatta cagttgtgct aaatgcccgt agtgagaaga 1620
acaggaacct tgagcatgta tagtagagga acctgcacag gtctgatggg tcagaggggt 1680
cttctctggg tttcactgag gatgagaagt aagcaaactg tggaaacatg caaaggaaaa 1740
agtgatagaa taatattcaa gacaaaaaga acagtatgag gcaagagaaa taatatgtat 1800
ttaaaatttt tggttactca atatcttata cttagtatga gtcctaaaat taaaaatgtg 1860
aaactgttgt actatacgta taacctaacc ttaattattc tgtaagaaca tgcttccata 1920
ggaaatagtg gataattttc agctatttaa ggcaaaagct aaaatagttc actcctcaac 1980
tgagacccaa agaattatag atatttttca tgatgaccca tgaaaaatat cactcatcta 2040
cataaaggag agactatatc tattttatag agaagctaag aaatatacct acacaaactt 2100
gtcaggtgct ttacaactac atagtacttt ttaacaacaa aataataatt ttaagaatga 2160
aaaatttaat catcgggaag aacgtcccac tacagacttc ctatcactgg cagttatatt 2220
tttgagcgta aaagggtcgt caaacgctaa atctaagtaa cgaattgaaa gtttaaagag 2280
ggggaagagt tggtttgcaa aggaaaagtt taaatagctt aatatcaata gaatgatcct 2340
gaagacagaa aaaactttgt cactcttcct ctctcatttt ctttctctct ctctcccctt 2400
ctcatacaca tgcctccccc accaaagaat ataatgtaaa ttaaatccac taaaatgtaa 2460
tggcatgaaa atctctgtag tctgaatcac taatattcct gagtttttat gagctcctag 2520
tacagctaaa gtttgcctat gcatgatcat ctatgcgtca gagcttcctc cttctacaag 2580
ctaactccct gcatctgggc atcaggactg ctccatacat ttgctgaaaa cttcttgtat 2640
ttcctgatgt aaaattgtgc aaacacctac aataaagcca tctactttta gggaaaggga 2700
gttgaaaatg caaccaactc ttggcgaact gtacaaacaa atctttgcta tactttattt 2760
caaataaatt ctttttaaaa taaaaaaaaa aaaaaaaaaa 2800
<210>251
<211>418
<212>PRT
<213>Homo sapiens
<220>
<223>trypsin-like enzyme，respiratory
<400>251
Met Tyr Arg Pro Ala Arg Val Thr Ser Thr Ser Arg Phe Leu Asn Pro
1 5 10 15
Tyr Val Val Cys Phe Ile Val Val Ala Gly Val Val Ile Leu Ala Val
20 25 30
Thr Ile Ala Leu Leu Val Tyr Phe Leu Ala Phe Asp Gln Lys Ser Tyr
35 40 45
Phe Tyr Arg Ser Ser Phe Gln Leu Leu Asn Val Glu Tyr Asn Ser Gln
50 55 60
Leu Asn Ser Pro Ala Thr Gln Glu Tyr Arg Thr Leu Ser Gly Arg Ile
65 70 75 80
Glu Ser Leu Ile Thr Lys Thr Phe Lys Glu Ser Asn Leu Arg Asn Gln
85 90 95
Phe Ile Arg Ala His ValAla Lys Leu Arg Gln Asp Gly Ser Gly Val
100 105 110
Arg Ala Asp Val Val Met Lys Phe Gln Phe Thr Arg Asn Asn Asn Gly
115 120 125
Ala Ser Met Lys Ser Arg Ile Glu Ser Val Leu Arg Gln Met Leu Asn
130 135 140
Asn Ser Gly Asn Leu Glu Ile Asn Pro Ser Thr Glu Ile Thr Ser Leu
145 150 155 160
Thr Asp Gln Ala Ala Ala Asn Trp Leu Ile Asn Glu Cys Gly Ala Gly
165 170 175
Pro Asp Leu Ile Thr Leu Ser Glu Gln Arg Ile Leu Gly Gly Thr Glu
180 185 190
Ala Glu Glu Gly Ser Trp Pro Trp Gln Val Ser Leu Arg Leu Asn Asn
195 200 205
Ala His His Cys Gly Gly Ser Leu Ile Asn Asn Met Trp Ile Leu Thr
210 215 220
Ala Ala His Cys Phe Arg Ser Asn Ser Asn Pro Arg Asp Trp Ile Ala
225 230 235 240
Thr Ser Gly Ile Ser Thr Thr Phe Pro Lys Leu Arg Met Arg Val Arg
245 250 255
Asn Ile Leu Ile His Asn Asn Tyr Lys Ser Ala Thr His Glu Asn Asp
260 265 270
Ile Ala Leu Val Arg Leu Glu Asn Ser Val Thr Phe Thr Lys Asp Ile
275 280 285
His Ser Val Cys Leu Pro Ala Ala Thr Gln Asn Ile Pro Pro Gly Ser
290 295 300
Thr Ala Tyr Val Thr Gly Trp Gly Ala Gln Glu Tyr Ala Gly His Thr
305 310 315 320
Val Pro Glu Leu Arg Gln Gly Gln Val Arg Ile Ile Ser Asn Asp Val
325 330 335
Cys Asn Ala Pro His Ser Tyr Asn Gly Ala Ile Leu Ser Gly Met Leu
340 345 350
Cys Ala Gly Val Pro Gln Gly Gly Val Asp Ala Cys Gln Gly Asp Ser
355 360 365
Gly Gly Pro Leu Val Gln Glu Asp Ser Arg Arg Leu Trp Phe Ile Val
370 375 380
Gly Ile Val Ser Trp Gly Asp Gln Cys Gly Leu Pro Asp Lys Pro Gly
385 390 395 400
Val Tyr Thr Arg Val Thr Ala Tyr Leu Asp Trp Ile Arg Gln Gln Thr
405 410 415
Gly Ile
<210>252
<211>3149
<212>DNA
<213>Homo sapiens
<220>
<223>Matripase
<400>252
gacgcctgtg agacccgcga gcggcctcgg ggaccatggg gagcgatcgg gcccgcaagg 60
gcggaggggg cccgaaggac ttcggcgcgg gactcaagta caactcccgg cacgagaaag 120
tgaatggctt ggaggaaggc gtggagttcc tgccagtcaa caacgtcaag aaggtggaaa 180
agcatggccc ggggcgctgg gtggtgctgg cagccgtgct gatcggcctc ctcttggtct 240
tgctggggat cggcttcctg gtgtggcatt tgcagtaccg ggacgtgcgt gtccagaagg 300
tcttcaatgg ctacatgagg atcacaaatg agaattttgt ggatgcctac gagaactcca 360
actccactga gtttgtaagc ctggccagca aggtgaagga cgcgctgaag ctgctgtaca 420
gcggagtccc attcctgggc ccctaccaca aggagtcggc tgtgacggcc ttcagcgagg 480
gcagcgtcat cgcctactac tggtctgagt tcagcatccc gcagcacctg gtggaggagg 540
ccgagcgcgt catggccgag gagcgcgtag tcatgctgcc cccgcgggcg cgctccctga 600
agtcctttgt ggtcacctca gtggtggctt tccccacgga ctccaaaaca gtacagagga 660
cccaggacaa cagctgcagc tttggcctgc acgcccgcgg tgtggagctg atgcgcttca 720
ccacgcccgg cttccctgac agcccctacc ccgctcatgc ccgctgccag tgggccctgc 780
ggggggacgc cgactcagtg ctgagcctca ccttccgcag ctttgacctt gcgtcctgcg 840
acgagcgcgg cagcgacctg gtgacggtgt acaacaccct gagccccatg gagccccacg 900
ccctggtgca gttgtgtggc acctaccctc cctcctacaa cctgaccttc cactcctccc 960
agaacgtcct gctcatcaca ctgataacca acactgagcg gcggcatccc ggctttgagg 1020
ccaccttctt ccagctgcct aggatgagca gctgtggagg ccgcttacgt aaagcccagg 1080
ggacattcaa cagcccctac tacccaggcc actacccacc caacattgac tgcacatgga 1140
acattgaggt gcccaacaac cagcatgtga aggtgcgctt caaattcttc tacctgctgg 1200
agcccggcgt gcctgcgggc acctgcccca aggactacgt ggagatcaat ggggagaaat 1260
actgcggaga gaggtcccag ttcgtcgtca ccagcaacag caacaagatc acagttcgct 1320
tccactcaga tcagtcctac accgacaccg gcttcttagc tgaatacctc tcctacgact 1380
ccagtgaccc atgcccgggg cagttcacgt gccgcacggg gcggtgtatc cggaaggagc 1440
tgcgctgtga tggctgggcc gactgcaccg accacagcga tgagctcaac tgcagttgcg 1500
acgccggcca ccagttcacg tgcaagaaca agttctgcaa gcccctcttc tgggtctgcg 1560
acagtgtgaa cgactgcgga gacaacagcg acgagcaggg gtgcagttgt ccggcccaga 1620
ccttcaggtg ttccaatggg aagtgcctct cgaaaagcca gcagtgcaat gggaaggacg 1680
actgtgggga cgggtccgac gaggcctcct gccccaaggt gaacgtcgtc acttgtacca 1740
aacacaccta ccgctgcctc aatgggctct gcttgagcaa gggcaaccct gagtgtgacg 1800
ggaaggagga ctgtagcgac ggctcagatg agaaggactg cgactgtggg ctgcggtcat 1860
tcacgagaca ggctcgtgtt gttgggggca cggatgcgga tgagggcgag tggccctggc 1920
aggtaagcct gcatgctctg ggccagggcc acatctgcgg tgcttccctc atctctccca 1980
actggctggt ctctgccgca cactgctaca tcgatgacag aggattcagg tactcagacc 2040
ccacgcagtg gacggccttc ctgggcttgc acgaccagag ccagcgcagc gcccctgggg 2100
tgcaggagcg caggctcaag cgcatcatct cccacccctt cttcaatgac ttcaccttcg 2160
actatgacat cgcgctgctg gagctggaga aaccggcaga gtacagctcc atggtgcggc 2220
ccatctgcct gccggacgcc tcccatgtct tccctgccgg caaggccatc tgggtcacgg 2280
gctggggaca cacccagtat ggaggcactg gcgcgctgat cctgcaaaag ggtgagatcc 2340
gcgtcatcaa ccagaccacc tgcgagaacc tcctgccgca gcagatcacg ccgcgcatga 2400
tgtgcgtggg cttcctcagc ggcggcgtgg actcctgcca gggtgattcc gggggacccc 2460
tgtccagcgt ggaggcggat gggcggatct tccaggccgg tgtggtgagc tggggagacg 2520
gctgcgctca gaggaacaag ccaggcgtgt acacaaggct ccctctgttt cgggactgga 2580
tcaaagagaa cactggggta taggggccgg ggccacccaa atgtgtacac ctgcggggcc 2640
acccatcgtc caccccagtg tgcacgcctg caggctggag actggaccgc tgactgcacc 2700
agcgccccca gaacatacac tgtgaactca atctccaggg ctccaaatct gcctagaaaa 2760
cctctcgctt cctcagcctc caaagtggag ctgggaggta gaaggggagg acactggtgg 2820
ttctactgac ccaactgggg gcaaaggttt gaagacacag cctcccccgc cagccccaag 2880
ctgggccgag gcgcgtttgt gtatatctgc ctcccctgtc tgtaaggagc agcgggaacg 2940
gagcttcgga gcctcctcag tgaaggtggt ggggctgccg gatctgggct gtggggccct 3000
tgggccacgc tcttgaggaa gcccaggctc ggaggaccct ggaaaacaga cgggtctgag 3060
actgaaaatg gtttaccagc tcccaggtga cttcagtgtg tgtattgtgt aaatgagtaa 3120
aacattttat ttctttttaa aaaaaaaaa 3149
<210>253
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Matripase
<400>253
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala ValThr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>254
<211>1258
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK13
<400>254
tcaggccccg cccgccctgc cctcccctcc cgatcccgga gccatgtggc ccctggccct 60
agtgatcgcc tccctgacct tggccttgtc aggaggtgtc tcccaggagt cttccaaggt 120
tctcaacacc aatgggacca gtgggtttct cccaggtggc tacacctgct tcccccactc 180
tcagccctgg caggctgccc tactagtgca agggcggcta ctctgtgggg gagtcctggt 240
ccaccccaaa tgggtcctca ctgccgcaca ctgtctaaag gaggggctca aagtttacct 300
aggcaagcac gccctagggc gtgtggaagc tggtgagcag gtgagggaag ttgtccactc 360
tatcccccac cctgaatacc ggagaagccc cacccacctg aaccacgacc atgacatcat 420
gcttctggag ctgcagtccc cggtccagct cacaggctac atccaaaccc tgcccctttc 480
ccacaacaac cgcctaaccc ctggcaccac ctgtcgggtg tctggctggg gcaccaccac 540
cagcccccag gtgaattacc ccaaaactct acaatgtgcc aacatccaac ttcgctcaga 600
tgaggagtgt cgtcaagtct acccaggaaa gatcactgac aacatgttgt gtgccggcac 660
aaaagagggt ggcaaagact cctgtgaggg tgactctggg ggccccctgg tctgtaacag 720
aacactgtat ggcatcgtct cctggggaga cttcccatgt gggcaacctg accggcctgg 780
tgtctacacc cgtgtctcaa gatacgtcct gtggatccgt gaaacaatcc gaaaatatga 840
aacccagcag caaaaatggt tgaagggccc acaataaaag ttgagaaatg taccggcttc 900
catcctgtca ccatgacttc ctcacatggt ctgcttagcc cttctctgct ccttattccc 960
agtgttccat ttgaaccagt gatccatgtc ctgaaaaatg ctcaatctca gctaacattc 1020
catgtttcag aagcattcag gcactgccag gcttgcagtc tcccagatgt tgcatccctg 1080
aaacatctca acaacctgaa tgtcccaacc cagacaatgg cccaggtctc tcaacttcat 1140
cagtgtggct tctatgagcc cagatcacca cctgaacgtt ctgtctgtgg cacattctta 1200
aatatttcca tcagcccatc tcaacaatat atgtcctata aatggaccat ccttgaca 1258
<210>255
<211>277
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hK13
<400>255
Met Trp Pro Leu Ala Leu Val Ile Ala Ser Leu Thr Leu Ala Leu Ser
1 5 10 15
Gly Gly Val Ser Gln Glu Ser Ser Lys Val Leu Asn Thr Asn Gly Thr
20 25 30
Ser Gly Phe Leu Pro Gly Gly Tyr Thr Cys Phe Pro His Ser Gln Pro
35 40 45
Trp Gln Ala Ala Leu Leu Val Gln Gly Arg Leu Leu Cys Gly Gly Val
50 55 60
Leu Val His Pro Lys Trp Val Leu Thr Ala Ala His Cys Leu Lys Glu
65 70 75 80
Gly Leu Lys Val Tyr Leu Gly Lys His Ala Leu Gly Arg Val Glu Ala
85 90 95
Gly Glu Gln Val Arg Glu Val Val His Ser Ile Pro His Pro Glu Tyr
100 105 110
Arg Arg Ser Pro Thr His Leu Asn His Asp His Asp Ile Met Leu Leu
115 120 125
Glu Leu Gln Ser Pro Val Gln Leu Thr Gly Tyr Ile Gln Thr Leu Pro
130 135 140
Leu Ser His Asn Asn Arg Leu Thr Pro Gly Thr Thr Cys Arg Val Ser
145 150 155 160
Gly Trp Gly Thr Thr Thr Ser Pro Gln Val Asn Tyr Pro Lys Thr Leu
165 170 175
Gln Cys Ala Asn Ile Gln Leu Arg Ser Asp Glu Glu Cys Arg Gln Val
180 185 190
Tyr Pro Gly Lys Ile Thr Asp Asn Met Leu Cys Ala Gly Thr Lys Glu
195 200 205
Gly Gly Lys Asp Ser Cys Glu Gly Asp Ser Gly Gly Pro Leu Val Cys
210 215 220
Asn Arg Thr Leu Tyr Gly Ile Val Ser Trp Gly Asp Phe Pro Cys Gly
225 230 235 240
Gln Pro Asp Arg Pro Gly Val Tyr Thr Arg Val Ser Arg Tyr Val Leu
245 250 255
Trp Ile Arg Glu Thr Ile Arg Lys Tyr Glu Thr Gln Gln Gln Lys Trp
260 265 270
Leu Lys Gly Pro Gln
275
<210>256
<211>1438
<212>DNA
<213>Homo sapiens
<220>
<223>kallikrein hK9
<400>256
catggaggag gaaggagatg gcatggctta ccataaagaa gcactggacg ccgggtgcac 60
gttccaggat ccaggtgccc aggggtcatg aagctgggac tcctctgtgc tctgctctct 120
ctgctggcag ggcatggctg ggcagacacc cgtgccatcg gggccgagga atgtcgcccc 180
aactcccagc cttggcaggc cggcctcttc caccttactc ggctcttctg tggggcgacc 240
ctcatcagtg accgctggct gctcacagct gcccactgcc gcaagccgta tctgtgggtc 300
cgccttggag agcaccacct ctggaaatgg gagggtccgg agcagctgtt ccgggttacg 360
gacttcttcc cccaccctgg cttcaacaag gacctcagcg ccaatgacca caatgatgac 420
atcatgctga tccgcctgcc caggcaggca cgtctgagtc ctgctgtgca gcccctcaac 480
ctcagccaga cctgtgtctc cccaggcatg cagtgtctca tctcaggctg gggggccgtg 540
tccagcccca aggcgctgtt tccagtcaca ctgcagtgtg ccaacatcag catcctggag 600
aacaaactct gtcactgggc ataccctgga cacatctcgg acagcatgct ctgtgcgggc 660
ctgtgggagg ggggccgagg ttcctgccag ggtgactctg ggggccccct ggtttgcaat 720
ggaaccttgg caggcgtggt gtctgggggt gctgagccct gctccagacc ccggcgcccc 780
gcagtctaca ccagcgtatg ccactacctt gactggatcc aagaaatcat ggagaactga 840
gcccgcgcgc cacgggggca ccttggaaga ccaagagagg ccgaagggca cggggtaggg 900
ggttctcgta gggtcccagc ctcaatggtt cccgccctgg acctccagct gccctgactc 960
ccctctggac actaagactc cgcccctgag gctccgcccc ctcacgaggt caagcaagac 1020
acagtcgcgc cccctcggaa cggagcaggg acacgccctt cagagcccgt ctctatgacg 1080
tcaccgacag ccatcacctc cttcttggaa cagcacagcc tgtggctccg ccccaaggaa 1140
ccacttacac aaaatagctc cgcccctcgg aactttgccc agtgggactt cccctcggga 1200
ctccacccct tgtggccccg cctccttcac cagagatctc gcccctcgtg atgtcagggg 1260
cgcagtagct ccgcccacgt ggagctcggg cggtgtagag ctcagcccct tgtggccccg 1320
tcctgggcgt gtgctgggtt tgaatcctgg cggagacctg gggggaaatt gagggagggt 1380
ctggatacct ttagagccaa tgcaacggat gatttttcag taaacgcggg aaacctca 1438
<210>257
<211>250
<212>PRT
<213>Homo sapiens
<220>
<223>kallikrein hK9
<400>257
Met Lys Leu Gly Leu Leu Cys Ala Leu Leu Ser Leu Leu Ala Gly His
1 5 10 15
Gly Trp Ala Asp Thr Arg Ala Ile Gly Ala Glu Glu Cys Arg Pro Asn
20 25 30
Ser Gln Pro Trp Gln Ala Gly Leu Phe His Leu Thr Arg Leu Phe Cys
35 40 45
Gly Ala Thr Leu Ile Ser Asp Arg Trp Leu Leu Thr Ala Ala His Cys
50 55 60
Arg Lys Pro Tyr Leu Trp Val Arg Leu Gly Glu His His Leu Trp Lys
65 70 75 80
Trp Glu Gly Pro Glu Gln Leu Phe Arg Val Thr Asp Phe Phe Pro His
85 90 95
Pro Gly Phe Asn Lys Asp Leu Ser Ala Asn Asp His Asn Asp Asp Ile
100 105 110
Met Leu Ile Arg Leu Pro Arg Gln Ala Arg Leu Ser Pro Ala Val Gln
115 120 125
Pro Leu Asn Leu Ser Gln Thr Cys Val Ser Pro Gly Met Gln Cys Leu
130 135 140
Ile Ser Gly Trp Gly Ala Val Ser Ser Pro Lys Ala Leu Phe Pro Val
145 150 155 160
Thr Leu Gln Cys Ala Asn Ile Ser Ile Leu Glu Asn Lys Leu Cys His
165 170 175
Trp Ala Tyr Pro Gly His Ile Ser Asp Ser Met Leu Cys Ala Gly Leu
180 185 190
Trp Glu Gly Gly Arg Gly Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu
195 200 205
Val Cys Asn Gly Thr Leu Ala Gly Val Val Ser Gly Gly Ala Glu Pro
210 215 220
Cys Ser Arg Pro Arg Arg Pro Ala Val Tyr Thr Ser Val Cys His Tyr
225 230 235 240
Leu Asp Trp Ile Gln Glu Ile Met Glu Asn
245 250
<210>258
<211>3212
<212>DNA
<213>Homo sapiens
<220>
<223>Mername-AA035 peptidase
<400>258
cttgagccag acccagtcca gctctggtgc ctgccctctg gtgcgagctg acctgagatg 60
cacttccctc ctctgtgagc tgtctcggca cccacttgca gtcactgccg cctgatgttg 120
ttactcttcc actccaaaag gatgcccgtg gccgaggccc cccaggtggc tggcgggcag 180
ggggacggag gtgatggcga ggaagcggag ccggagggga tgttcaaggc ctgtgaggac 240
tccaagagaa aagcccgggg ctacctccgc ctggtgcccc tgtttgtgct gctggccctg 300
ctcgtgctgg cttcggcggg ggtgctactc tggtatttcc tagggtacaa ggcggaggtg 360
atggtcagcc aggtgtactc aggcagtctg cgtgtactca atcgccactt ctcccaggat 420
cttacccgcc gggaatctag tgccttccgc agtgaaaccg ccaaagccca gaagatgctc 480
aaggagctca tcaccagcac ccgcctggga acttactaca actccagctc cgtctattcc 540
tttggggagg gacccctcac ctgcttcttc tggttcattc tccaaatccc cgagcaccgc 600
cggctgatgc tgagccccga ggtggtgcag gcactgctgg tggaggagct gctgtccaca 660
gtcaacagct cggctgccgt cccctacagg gccgagtacg aagtggaccc cgagggccta 720
gtgatcctgg aagccagtgt gaaagacata gctgcattga attccacgct gggttgttac 780
cgctacagct acgtgggcca gggccaggtc ctccggctga aggggcctga ccacctggcc 840
tccagctgcc tgtggcacct gcagggcccc aaggacctca tgctcaaact ccggctggag 900
tggacgctgg cagagtgccg ggaccgactg gccatgtatg acgtggccgg gcccctggag 960
aagaggctca tcacctcggt gtacggctgc agccgccagg agcccgtggt ggaggttctg 1020
gcgtcggggg ccatcatggc ggtcgtctgg aagaagggcc tgcacagcta ctacgacccc 1080
ttcgtgctct ccgtgcagcc ggtggtcttc caggcctgtg aagtgaacct gacgctggac 1140
aacaggctcg actcccaggg cgtcctcagc accccgtact tccccagcta ctactcgccc 1200
caaacccact gctcctggca cctcacggtg ccctctctgg actacggctt ggccctctgg 1260
tttgatgcct atgcactgag gaggcagaag tatgatttgc cgtgcaccca gggccagtgg 1320
acgatccaga acaggaggct gtgtggcttg cgcatcctgc agccctacgc cgagaggatc 1380
cccgtggtgg ccacggccgg gatcaccatc aacttcacct cccagatctc cctcaccggg 1440
cccggtgtgc gggtgcacta tggcttgtac aaccagtcgg acccctgccc tggagagttc 1500
ctctgttctg tgaatggact ctgtgtccct gcctgtgatg gggtcaagga ctgccccaac 1560
ggcctggatg agagaaactg cgtttgcaga gccacattcc agtgcaaaga ggacagcaca 1620
tgcatctcac tgcccaaggt ctgtgatggg cagcctgatt gtctcaacgg cagcgacgaa 1680
gagcagtgcc aggaaggggt gccatgtggg acattcacct tccagtgtga ggaccggagc 1740
tgcgtgaaga agcccaaccc gcagtgtgat gggcggcccg actgcaggga cggctcggat 1800
gaggagcact gtgactgtgg cctccagggc ccctccagcc gcattgttgg tggagctgtg 1860
tcctccgagg gtgagtggcc atggcaggcc agcctccagg ttcggggtcg acacatctgt 1920
gggggggccc tcatcgctga ccgctgggtg ataacagctg cccactgctt ccaggaggac 1980
agcatggcct ccacggtgct gtggaccgtg ttcctgggca aggtgtggca gaactcgcgc 2040
tggcctggag aggtgtcctt caaggtgagc cgcctgctcc tgcacccgta ccacgaagag 2100
gacagccatg actacgacgt ggcgctgctg cagctcgacc acccggtggt gcgctcggcc 2160
gccgtgcgcc ccgtctgcct gcccgcgcgc tcccacttct tcgagcccgg cctgcactgc 2220
tggattacgg gctggggcgc cttgcgcgag ggcggcccca tcagcaacgc tctgcagaaa 2280
gtggatgtgc agttgatccc acaggacctg tgcagcgagg tctatcgcta ccaggtgacg 2340
ccacgcatgc tgtgtgccgg ctaccgcaag ggcaagaagg atgcctgtca gggtgactca 2400
ggtggtccgc tggtgtgcaa ggcactcagt ggccgctggt tcctggcggg gctggtcagc 2460
tggggcctgg gctgtggccg gcctaactac ttcggcgtct acacccgcat cacaggtgtg 2520
atcagctgga tccagcaagt ggtgacctga ggaactgccc ccctgcaaag cagggcccac 2580
ctcctggact cagagagccc agggcaactg ccaagcaggg ggacaagtat tctggcgggg 2640
ggtgggggag agagcaggcc ctgtggtggc aggaggtggc atcttgtctc gtccctgatg 2700
tctgctccag tgatggcagg aggatggaga agtgccagca gctgggggtc aagacgtccc 2760
ctgaggaccc aggcccacac ccagcccttc tgcctcccaa ttctctctcc tccgtcccct 2820
tcctccactg ctgcctaatg caaggcagtg gctcagcagc aagaatgctg gttctacatc 2880
ccgaggagtg tctgaggtgc gccccactct gtacagaggc tgtttgggca gccttgcctc 2940
cagagagcag attccagctt cggaagcccc tggtctaact tgggatctgg gaatggaagg 3000
tgctcccatc ggaggggacc ctcagagccc tggagactgc caggtgggcc tgctgccact 3060
gtaagccaaa aggtggggaa gtcctgactc cagggtcctt gccccacccc tgcctgccac 3120
ctgggccctc acagcccaga ccctcactgg gaggtgagct cagctgccct ttggaataaa 3180
gctgcctgat caaaaaaaaa aaaaaaaaaa aa 3212
<210>259
<211>811
<212>PRT
<213>Homo sapiens
<220>
<223>Mername-AA035 peptidase
<400>259
Met Leu Leu Leu Phe His Ser Lys Arg Met Pro Val Ala Glu Ala Pro
1 5 10 15
Gln Val Ala Gly Gly Gln Gly Asp Gly Gly Asp Gly Glu Glu Ala Glu
20 25 30
Pro Glu Gly Met Phe Lys Ala Cys Glu Asp Ser Lys Arg Lys Ala Arg
35 40 45
Gly Tyr Leu Arg Leu Val Pro Leu Phe Val Leu Leu Ala Leu Leu Val
50 55 60
Leu Ala Ser Ala Gly Val Leu Leu Trp Tyr Phe Leu Gly Tyr Lys Ala
65 70 75 80
Glu Val Met Val Ser Gln Val Tyr Ser Gly Ser Leu Arg Val Leu Asn
85 90 95
Arg His Phe Ser Gln Asp Leu Thr Arg Arg Glu Ser Ser Ala Phe Arg
100 105 110
Ser Glu Thr Ala Lys Ala Gln Lys Met Leu Lys Glu Leu Ile Thr Ser
115 120 125
Thr Arg Leu Gly Thr Tyr Tyr Asn Ser Ser Ser Val Tyr Ser Phe Gly
130 135 140
Glu Gly Pro Leu Thr Cys Phe Phe Trp Phe Ile Leu Gln Ile Pro Glu
145 150 155 160
His Arg Arg Leu Met Leu Ser Pro Glu Val Val Gln Ala Leu Leu Val
165 170 175
Glu Glu Leu Leu Ser Thr Val Asn Ser Ser Ala Ala Val Pro Tyr Arg
180 185 190
Ala Glu Tyr Glu Val Asp Pro Glu Gly Leu Val Ile Leu Glu Ala Ser
195 200 205
Val Lys Asp Ile Ala Ala Leu Asn Ser Thr Leu Gly Cys Tyr Arg Tyr
210 215 220
Ser Tyr Val Gly Gln Gly Gln Val Leu Arg Leu Lys Gly Pro Asp His
225 230 235 240
Leu Ala Ser Ser Cys Leu Trp His Leu Gln Gly Pro Lys Asp Leu Met
245 250 255
Leu Lys Leu Arg Leu Glu Trp Thr Leu Ala Glu Cys Arg Asp Arg Leu
260 265 270
Ala Met Tyr Asp Val Ala Gly Pro Leu Glu Lys Arg Leu Ile Thr Ser
275 280 285
Val Tyr Gly Cys Ser Arg Gln Glu Pro Val Val Glu Val Leu Ala Ser
290 295 300
Gly Ala Ile Met Ala Val Val Trp Lys Lys Gly Leu His Ser Tyr Tyr
305 310 315 320
Asp Pro Phe Val Leu Ser Val Gln Pro Val Val Phe Gln Ala Cys Glu
325 330 335
Val Asn Leu Thr Leu Asp Asn Arg Leu Asp Ser Gln Gly Val Leu Ser
340 345 350
Thr Pro Tyr Phe Pro Ser Tyr Tyr Ser Pro Gln Thr His Cys Ser Trp
355 360 365
His Leu Thr Val Pro Ser Leu Asp Tyr Gly Leu Ala Leu Trp Phe Asp
370 375 380
Ala Tyr Ala Leu Arg Arg Gln Lys Tyr Asp Leu Pro Cys Thr Gln Gly
385 390 395 400
Gln Trp Thr Ile Gln Asn Arg Arg Leu Cys Gly Leu Arg Ile Leu Gln
405 410 415
Pro Tyr Ala Glu Arg Ile Pro Val Val Ala Thr Ala Gly Ile Thr Ile
420 425 430
Asn Phe Thr Ser Gln Ile Ser Leu Thr Gly Pro Gly Val Arg Val His
435 440 445
Tyr Gly Leu Tyr Asn Gln Ser Asp Pro Cys Pro Gly Glu Phe Leu Cys
450 455 460
Ser Val Asn Gly Leu Cys Val Pro Ala Cys Asp Gly Val Lys Asp Cys
465 470 475 480
Pro Asn Gly Leu Asp Glu Arg Asn Cys Val Cys Arg Ala Thr Phe Gln
485 490 495
Cys Lys Glu Asp Ser Thr Cys Ile Ser Leu Pro Lys Val Cys Asp Gly
500 505 510
Gln Pro Asp Cys Leu Asn Gly Ser Asp Glu Glu Gln Cys Gln Glu Gly
515 520 525
Val Pro Cys Gly Thr Phe Thr Phe Gln Cys Glu Asp Arg Ser Cys Val
530 535 540
Lys Lys Pro Asn Pro Gln Cys Asp Gly Arg Pro Asp Cys Arg Asp Gly
545 550 555 560
Ser Asp Glu Glu His Cys Asp Cys Gly Leu Gln Gly Pro Ser Ser Arg
565 570 575
Ile Val Gly Gly Ala Val Ser Ser Glu Gly Glu Trp Pro Trp Gln Ala
580 585 590
Ser Leu Gln Val Arg Gly Arg His Ile Cys Gly Gly Ala Leu Ile Ala
595 600 605
Asp Arg Trp Val Ile Thr Ala Ala His Cys Phe Gln Glu Asp Ser Met
610 615 620
Ala Ser Thr Val Leu Trp Thr Val Phe Leu Gly Lys Val Trp Gln Asn
625 630 635 640
Ser Arg Trp Pro Gly Glu Val Ser Phe Lys ValSer Arg Leu Leu Leu
645 650 655
His Pro Tyr His Glu Glu Asp Ser His Asp Tyr Asp Val Ala Leu Leu
660 665 670
Gln Leu Asp His Pro Val Val Arg Ser Ala Ala Val Arg Pro Val Cys
675 680 685
Leu Pro Ala Arg Ser His Phe Phe Glu Pro Gly Leu His Cys Trp Ile
690 695 700
Thr Gly Trp Gly Ala Leu Arg Glu Gly Gly Pro Ile Ser Asn Ala Leu
705 710 715 720
Gln Lys Val Asp Val Gln Leu Ile Pro Gln Asp Leu Cys Ser Glu Val
725 730 735
Tyr Arg Tyr Gln Val Thr Pro Arg Met Leu Cys Ala Gly Tyr Arg Lys
740 745 750
Gly Lys Lys Asp Ala Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys
755 760 765
Lys Ala Leu Ser Gly Arg Trp Phe Leu Ala Gly Leu Val Ser Trp Gly
770 775 780
Leu Gly Cys Gly Arg Pro Asn Tyr Phe Gly Val Tyr Thr Arg Ile Thr
785 790 795 800
Gly gal Ile Ser Trp Ile Gln Gln Val Val Thr
805 810
<210>260
<211>3750
<212>DNA
<213>Homo sapiens
<220>
<223>umbilical vein proteinase
<400>260
gcgctgctcg ccagcttgct cgcactcggc tgtgcggcgg ggcaggcatg ggagccgcgc 60
gctctctccc ggcgcccaca cctgtctgag cggcgcagcg agccgcggcc cgggcgggct 120
gctcggcgcg gaacagtgct cggcatggca gggattccag ggctcctctt ccttctcttc 180
tttctgctct gtgctgttgg gcaagtgagc ccttacagtg ccccctggaa acccacttgg 240
cctgcatacc gcctccctgt cgtcttgccc cagtctaccc tcaatttagc caagccagac 300
tttggagccg aagccaaatt agaagtatct tctteatgtg gaccccagtg tcataaggga 360
actccactgc ccacttacga agaggccaag caatatctgt cttatgaaac gctctatgcc 420
aatggcagcc gcacagagac gcaggtgggc atctacatcc tcagcagtag tggagatggg 480
gcccaacacc gagactcagg gtcttcagga aagtctcgaa ggaagcggca gatttatggc 540
tatgacagca ggttcagcat ttttgggaag gacttcctgc tcaactaccc tttctcaaca 600
tcagtgaagt tatccacggg ctgcaccggc accctggtgg cagagaagca tgtcctcaca 660
gctgcccact gcatacacga tggaaaaacc tatgtgaaag gaacccagaa gcttcgagtg 720
ggcttcctaa agcccaagtt taaagatggt ggtcgagggg ccaacgactc cacttcagcc 780
atgcccgagc agatgaaatt tcagtggatc cgggtgaaac gcacccatgt gcccaagggt 840
tggatcaagg gcaatgccaa tgacatcggc atggattatg attatgccct cctggaactc 900
aaaaagcccc acaagagaaa atttatgaag attggggtga gccctcctgc taagcagctg 960
ccagggggca gaattcactt ctctggttat gacaatgacc gaccaggcaa tttggtgtat 1020
cgcttctgtg acgtcaaaga cgagacctat gacttgctct accagcaatg cgatgcccag 1080
ccaggggcca gcgggtctgg ggtctatgtg aggatgtgga agagacagca gcagaagtgg 1140
gagcgaaaaa ttattggcat tttttcaggg caccagtggg tggacatgaa tggttcccca 1200
caggatttca acgtggctgt cagaatcact cctctcaaat atgcccagat ttgctattgg 1260
attaaaggaa actacctgga ttgtagggag gggtgacaca gtgttccctc ctggcagcaa 1320
ttaagggtct tcatgttctt attttaggag aggccaaatt gttttttgtc attggcgtgc 1380
acacgtgtgt gtgtgtgtgt gtgtgtgtgt aaggtgtctt ataatctttt acctatttct 1440
tacaattgca agatgactgg ctttactatt tgaaaactgg tttgtgtatc atatcatata 1500
tcatttaagc agtttgaagg catacttttg catagaaata aaaaaaatac tgatttgggg 1560
caatgaggaa tatttgacaa ttaagttaat cttcacgttt ttgcaaactt tgatttttat 1620
ttcatctgaa cttgtttcaa agatttatat taaatatttg gcatacaaga gatatgaatt 1680
cttatatgtg tgcatgtgtg ttttcttctg agattcatct tggtggtggg tttttttgtt 1740
tttttaattc agtgcctgat ctttaatgct tccataaggc agtgttccca tttaggaact 1800
ttgacagcat ttgttaggca gaatattttg gatttggagg catttgcatg gtagtctttg 1860
aacagtaaaa tgatgtgttg actatactga tacacatatt aaactatacc ttatagtaaa 1920
ccagtatccc aagctgcttt tagttccaaa aatagtttct tttccaaagg ttgttgctct 1980
actttgtagg aagtctttgc atatggccct cccaacttta aagtcatacc agagtggcca 2040
agagtgttta tcccaaccct tccatttaac aggatttcac tcacatttct ggaactagct 2100
atttttcaga agacaataat cagggcttaa ttagaacagg ctgtatttcc tcccagcaaa 2160
cagttgtggc cacactaaaa acaatcatag cattttaccc ctggattata gcacatctca 2220
tgttttatca tttggatgga gtaatttaaa atgaattaaa ttccagagaa caatggaagc 2280
attgcctggc agatgtcaca acagaataac cacttgtttg gagcctggca cagtcctcca 2340
gcctgatcaa aaattattct gcatagtttt cagtgtgctt tctgggagct atgtacttct 2400
tcaatttgga aacttttctc tctcatttat agtgaaaata cttggaagtt actttaagaa 2460
aaccagtgtg gcctttttcc ctctagcttt aaaagggccg cttttgctgg aatgctctag 2520
gttatagata aacaattagg tataatagca aaaatgaaaa ttggaagaat gcaaaatgga 2580
tcagaatcat gccttccaat aaaggccttt acacatgttt tatcaatatg attatcaaat 2640
cacagcatat acagaaaaga cttggactta ttgtatgttt ttattttatg gctctcggcc 2700
taagcacttc tttctaaatg tatcggagaa aaaatcaaat ggactacaag cacgtgtttg 2760
ctgtgcttgc accccaggta aacctgcatt gtagcaattt gtaaggatat tcagatggag 2820
cactgtcact tagacattct ctgggggatt ttctgcttgt ctttcttgag ctttttggaa 2880
ggataattct gataaggcac tcaagaaacg tacaaccaca gtgctttctt caaatcatat 2940
gagaaatact atgcatagca aggagatgca gagccgccag gaaaattctg agttccagca 3000
caattttctt tggaatctaa caggaatcta gcctgaggaa gaagggaggt ctccatttct 3060
atgtctggta tttgggggtt ttgtttgttt ttgctttagc ttggtgaaaa aaagttcact 3120
gaacaccaag accagaatgg atttttttaa aaaaatagat gttccttttg tgaagcacct 3180
tgattccttg attttgattt tttgcaaagt tagacaatgg cacaaagtca aaatgaaatc 3240
aatgtttagt tcacaagtag atgtaattta ctaaagaatg atacacccat atgctatata 3300
cagcttaact cacagaactg taaaagaaaa ttataaaata attcaacatg tccatctttt 3360
tagtgataat aaaagaaagc atggtattaa actatcatag aagtagacag aaaaagaaaa 3420
aaggactcat ggcattatta atataattag tgctttacat gtgttagtta tacatattag 3480
aagcatattt gcctagtaag gctagtagaa ccacatttcc caaagtgtgc tccttaaaca 3540
ctcatgcctt atgattttct accaaaagta aaaagggttg tattaagtca gaggaagatg 3600
cctctccatt ttccctctct ttatcagagg ttcacatgcc tgtctgcaca ttaaaagctc 3660
tgggaagacc tgttgtaaag ggacaagttg aggttgtaaa atctgcattt aaataaacat 3720
ctttgatcac aaaaaaaaaa aaaaaaaaaa 3750
<210>261
<211>383
<212>PRT
<213>Homo sapiens
<220>
<223>umbilical vein proteinase
<400>261
Met Ala Gly Ile Pro Gly Leu Leu Phe Leu Leu Phe Phe Leu Leu Cys
1 5 10 15
Ala Val Gly Gln Val Ser Pro Tyr Ser Ala Pro Trp Lys Pro Thr Trp
20 25 30
Pro Ala Tyr Arg Leu Pro Val Val Leu Pro Gln Ser Thr Leu Asn Leu
35 40 45
Ala Lys Pro Asp Phe Gly Ala Glu Ala Lys Leu Glu Val Ser Ser Ser
50 55 60
Cys Gly Pro Gln Cys His Lys Gly Thr Pro Leu Pro Thr Tyr Glu Glu
65 70 75 80
Ala Lys Gln Tyr Leu Ser Tyr Glu Thr Leu Tyr Ala Asn Gly Ser Arg
85 90 95
Thr Glu Thr Gln Val Gly Ile Tyr Ile Leu Ser Ser Ser Gly Asp Gly
100 105 110
Ala Gln His Arg Asp Ser Gly Ser Ser Gly Lys Ser Arg Arg Lys Arg
115 120 125
Gln Ile Tyr Gly Tyr Asp Ser Arg Phe Ser Ile Phe Gly Lys Asp Phe
130 135 140
Leu Leu Asn Tyr Pro Phe Ser Thr Ser ValLys Leu Ser Thr Gly Cys
145 150 155 160
Thr Gly Thr Leu Val Ala Glu Lys His Val Leu Thr Ala Ala His Cys
165 170 175
Ile His Asp Gly Lys Thr Tyr Val Lys Gly Thr Gln Lys Leu Arg Val
180 185 190
Gly Phe Leu Lys Pro Lys Phe Lys Asp Gly Gly Arg Gly Ala Asn Asp
195 200 205
Ser Thr Ser Ala Met Pro Glu Gln Met Lys Phe Gln Trp Ile Arg Val
210 215 220
Lys Arg Thr His Val Pro Lys Gly Trp Ile Lys Gly Asn Ala Asn Asp
225 230 235 240
Ile Gly Met Asp Tyr Asp Tyr Ala Leu Leu Glu Leu Lys Lys Pro His
245 250 255
Lys Arg Lys Phe Met Lys Ile Gly Val Ser Pro Pro Ala Lys Gln Leu
260 265 270
Pro Gly Gly Arg Ile His Phe Ser Gly Tyr Asp Asn Asp Arg Pro Gly
275 280 285
Asn Leu Val Tyr Arg Phe Cys Asp Val Lys Asp Glu Thr Tyr Asp Leu
290 295 300
Leu Tyr Gln Gln Cys Asp Ala Gln Pro Gly Ala Ser Gly Ser Gly Val
305 310 315 320
Tyr Val Arg Met Trp Lys Arg Gln Gln Gln Lys Trp Glu Arg Lys Ile
325 330 335
Ile Gly Ile Phe Ser Gly His Gln Trp Val Asp Met Asn Gly Ser Pro
340 345 350
Gln Asp Phe Asn Val Ala Val Arg Ile Thr Pro Leu Lys Tyr Ala Gln
355 360 365
Ile Cys Tyr Trp Ile Lys Gly Asn Tyr Leu Asp Cys Arg Glu Gly
370 375 380
<210>262
<211>26
<212>PRT
<213>Homo sapiens
<220>
<223>LCLP proteinase (LCLP (N-terminus))
<400>262
Ile Ile Gly Gly Thr Glu Ser Lys Pro Asp Ser Arg Pro Tyr Met Ala
1 5 10 15
Leu Leu Gln Ile Val Glu Pro Ala Val His
20 25
<210>263
<211>2265
<212>DNA
<213>Homo sapiens
<220>
<223>Spinesin
<400>263
acgcgggata cagggagggg ccatgtgcga accagggaga cctcatcttc caaccaagct 60
tgctgggctt gcatttaatc aatgcatggc cagagaacag gagcggaaca ttgcctagta 120
gaccctgagg ctttacaaca gtgctactga cccctatgag cctgatgctg gatgaccaac 180
cccctatgga ggcccagtat gcagaggagg gcccaggacc tgggatcttc agagcagagc 240
ctggagacca gcagcatccc atttctcagg cggtgtgctg gcgttccatg cgacgtggct 300
gtgcagtgct gggagccctg gggctgctgg ccggtgcagg tgttggctca tggctcctag 360
tgctgtatct gtgtcctgct gcctctcagc ccatttccgg gaccttgcag gatgaggaga 420
taactttgag ctgctcagag gccagcgctg aggaagctct gctccctgca ctccccaaaa 480
cagtatcttt cagaataaac agcgaagact tcttgctgga agcgcaagtg agggatcagc 540
cacgctggct cctggtctgc catgagggct ggagccccgc cctggggctg cagatctgct 600
ggagccttgg gcatctcaga ctcactcacc acaagggagt aaacctcact gacatcaaac 660
tcaacagttc ccaggagttt gctcagctct ctcctagact gggaggcttc ctggaggagg 720
cgtggcagcc caggaacaac tgcacttctg gtcaagttgt ttccctcaga tgctctgagt 780
gtggagcgag gcccctggct tcccggatag ttggtgggca gtctgtggct cctgggcgct 840
ggccgtggca ggccagcgtg gccctgggct tccggcacac gtgtgggggc tctgtgctag 900
cgccacgctg ggtggtgact gctgcacatt gtatgcacag tttcaggctg gcccgcctgt 960
ccagctggcg ggttcatgcg gggctggtca gccacagtgc cgtcaggccccaccaagggg 1020
ctctggtgga gaggattatc ccacaccccc tctacagtgc ccagaatcat gactacgacg 1080
tcgccctcct gaggctccag accgctctca acttctcaga cactgtgggc gctgtgtgcc 1140
tgccggccaa ggaacagcat tttccgaagg gctcgcggtg ctgggtgtct ggctggggcc 1200
acacccaccc tagccatact tacagctcgg atatgctcca ggacacggtg gtgcccttgt 1260
tcagcactca gctctgcaac agctcttgcg tgtacagcgg agccctcacc ccccgcatgc 1320
tttgcgctgg ctacctggac ggaagggctg atgcatgcca gggagatagc gggggccccc 1380
tagtgtgccc agatggggac acatggcgcc tagtgggggt ggtcagctgg gggcgtgcgt 1440
gcgcagagcc caatcaccca ggtgtctacg ccaaggtagc tgagtttctg gactggatcc 1500
atgacactgc tcaggactcc ctcctctgag tcctgctgtt tcctccagtc tcactgcaca 1560
ccactgcctc atgcttcctg gggcctccag cagctccact aatggaggag aggcagtagc 1620
ctccgacaca gaacgcatgg acctcctact actgtgtgtg aggaacagtc actacccact 1680
ggccagccac ccagccaaca ggtctctcct cttgggccct gatttcagag tcctctttct 1740
cactagagac tcaatgacag aagagaggct gggacttggt tgggcatgct gtggttgctg 1800
agggatgagg gggaggagag aggtaggagc tggagatgaa gagactgcta gaagcagcag 1860
gaagcctgcc cttctgccct ctcccctccc tgcccctgtg tgagtctttt agggagggtg 1920
actgggaggt gccccccgtc ccaccttttt cctgtgctct aggtgggcta agtgcctccc 1980
tagaggactc catggctgag aggctcctgg gcagatgggg tcaaggctgg gccagtccca 2040
gatgaagcct atgggagtca ggaccctctc cactctccct ctccactccc cttcctgttc 2100
tcacctggct gtggctggcc ctgtgtgggg tgggtacact ggaaaacaag aaggttggag 2160
ttggtctagg acattggttt taaatgacag ttctgtgaac tggtccaagg aggttctgtt 2220
attaaagtga tatatggtct tgaaaaaaaa aaaaaaaaaa aaaaa 2265
<210>264
<211>457
<212>PRT
<213>Homo sapiens
<220>
<223>Spinesin
<400>264
Met Ser Leu Met Leu Asp Asp Gln Pro Pro Met Glu Ala Gln Tyr Ala
1 5 10 15
Glu Glu Gly Pro Gly Pro Gly Ile Phe Arg Ala Glu Pro Gly Asp Gln
20 25 30
Gln His Pro Ile Ser Gln Ala Val Cys Trp Arg Ser Met Arg Arg Gly
35 40 45
Cys Ala Val Leu Gly Ala Leu Gly Leu Leu Ala Gly Ala Gly Val Gly
50 55 60
Ser Trp Leu Leu Val Leu Tyr Leu Cys Pro Ala Ala Ser Gln Pro Ile
65 70 75 80
Ser Gly Thr Leu Gln Asp Glu Glu Ile Thr Leu Ser Cys Ser Glu Ala
85 90 95
Ser Ala Glu Glu Ala Leu Leu Pro Ala Leu Pro Lys Thr Val Ser Phe
100 105 110
Arg Ile Asn Ser Glu Asp Phe Leu Leu Glu Ala Gln Val Arg Asp Gln
115 120 125
Pro Arg Trp Leu Leu Val Cys His Glu Gly Trp Ser Pro Ala Leu Gly
130 135 140
Leu Gln Ile Cys Trp Ser Leu Gly His Leu Arg Leu Thr His His Lys
145 150 155 160
Gly Val Asn Leu Thr Asp Ile Lys Leu Asn Ser Ser Gln Glu Phe Ala
165 170 175
Gln Leu Ser Pro Arg Leu Gly Gly Phe Leu Glu Glu Ala Trp Gln Pro
180 185 190
Arg Asn Asn Cys Thr Ser Gly Gln Val Val Ser Leu Arg Cys Ser Glu
195 200 205
Cys Gly Ala Arg Pro Leu Ala Ser Arg Ile Val Gly Gly Gln Ser Val
210 215 220
Ala Pro Gly Arg Trp Pro Trp Gln Ala Ser Val Ala Leu Gly Phe Arg
225 230 235 240
His Thr Cys Gly Gly Ser Val Leu Ala Pro Arg Trp Val Val Thr Ala
245 250 255
Ala His Cys Met His Ser Phe Arg Leu Ala Arg Leu Ser Ser Trp Arg
260 265 270
Val His Ala Gly Leu Val Ser His Ser Ala Val Arg Pro His Gln Gly
275 280 285
Ala Leu Val Glu Arg Ile Ile Pro His Pro Leu Tyr Ser Ala Gln Asn
290 295 300
His Asp Tyr Asp Val Ala Leu Leu Arg Leu Gln Thr Ala Leu Asn Phe
305 310 315 320
Ser Asp Thr Val Gly Ala Val Cys Leu Pro Ala Lys Glu Gln His Phe
325 330 335
Pro Lys Gly Ser Arg Cys Trp Val Ser Gly Trp Gly His Thr His Pro
340 345 350
Ser His Thr Tyr Ser Ser Asp Met Leu Gln Asp Thr Val Val Pro Leu
355 360 365
Phe Ser Thr Gln Leu Cys Asn Ser Ser Cys Val Tyr Ser Gly Ala Leu
370 375 380
Thr Pro Arg Met Leu Cys Ala Gly Tyr Leu Asp Gly Arg Ala Asp Ala
385 390 395 400
Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Pro Asp Gly Asp Thr
405 410 415
Trp Arg Leu Val Gly Val Val Ser Trp Gly Arg Ala Cys Ala Glu Pro
420 425 430
Asn His Pro Gly Val Tyr Ala Lys Val Ala Glu Phe Leu Asp Trp Ile
435 440 445
His Asp Thr Ala Gln Asp Ser Leu Leu
450 455
<210>265
<211>1243
<212>DNA
<213>Homo sapiens
<220>
<223>Mername-AA178 peptidase
<400>265
atggctgccc ctgcttccgt catgggccca ctcgggccct ctgccctggg ccttctgctg 60
ctgctcctgg tggtggcccc tccccgggtc gcagcattgg tccacagaca gccagagaac 120
cagggaatct ccctaactgg cagcgtggcc tgtggtcggc ccagcatgga ggggaaaatc 180
ctgggcggcg tccctgcgcc cgagaggaag tggccgtggc aggtcagcgt gcactacgca 240
ggcctccacg tctgcggcgg ctccatcctc aatgagtact gggtgctgtc agctgcgcac 300
tgctttcaca gggacaagaa tatcaaaatc tatgacatgt acgtaggcct cgtaaacctc 360
agggtggccg gcaaccacac ccagtggtat gaggtgaaca gggtgatcct gcaccccaca 420
tatgagatgt accaccccat cggaggtgac gtggccctgg tgcagctgaa gacccgcatt 480
gtgttttctg agtccgtgct cccggtttgc cttgcaactc cagaagtgaa ccttaccagt 540
gccaattgct gggctacggg atggggacta gtctcaaaac aaggtgagac ctcagacgag 600
ctgcaggaga tgcagctccc gctgatcctg gagccctggt gccacctgct ctacggacac 660
atgtcctaca tcatgcccga catgctgtgt gctggggaca tcctgaatgc taagaccgtg 720
tgtgagggcg actccggggg cccacttgtc tgtgaattca accgcagctg gttgcagatt 780
ggaattgtga gctggggccg aggctgctcc aaccctctgt accctggagt gtatgccagt 840
gtttcctatt tctcaaaatg gatatgtgat aacatagaaa tcacgcccac tcctgctcag 900
ccagcccctg ctctctctcc agctctgggg cccactctca gcgtcctaat ggccatgctg 960
gctggctggt cagtgctgtg aggtcaggat acccactcta ggattctcat ggctgcacac 1020
cctgccccag cccagctgcc tccagacccc taagcatctc ctgtcctggc ctctctgaag 1080
cagacaaggg ccacctatcc cgggggtgga tgctgagtcc aggaggtgat gagcaagtgt 1140
acaaaagaaa aaagggaagg gggagagggg ctggtcaggg agaacccagc ttgggcagag 1200
tgcacctgag atttgataag atcattaaat atttacaaag caa 1243
<210>266
<211>326
<212>PRT
<213>Homo sapiens
<220>
<223>Mername-AA178 peptidase
<400>266
Met Ala Ala Pro Ala Ser Val Met Gly Pro Leu Gly Pro Ser Ala Leu
1 5 10 15
Gly Leu Leu Leu Leu Leu Leu Val Val Ala Pro Pro Arg Val Ala Ala
20 25 30
Leu Val His Arg Gln Pro Glu Asn Gln Gly Ile Ser Leu Thr Gly Ser
35 40 45
Val Ala Cys Gly Arg Pro Ser Met Glu Gly Lys Ile Leu Gly Gly Val
50 55 60
Pro Ala Pro Glu Arg Lys Trp Pro Trp Gln Val Ser Val His Tyr Ala
65 70 75 80
Gly Leu His Val Cys Gly Gly Ser Ile Leu Asn Glu Tyr Trp Val Leu
85 90 95
Ser Ala Ala His Cys Phe His Arg Asp Lys Asn Ile Lys Ile Tyr Asp
100 105 110
Met Tyr Val Gly Leu Val Asn Leu Arg Val Ala Gly Asn His Thr Gln
115 120 125
Trp Tyr Glu Val Asn Arg Val Ile Leu His Pro Thr Tyr Glu Met Tyr
130 135 140
His Pro Ile Gly Gly Asp Val Ala Leu Val Gln Leu Lys Thr Arg Ile
145 150 155 160
Val Phe Ser Glu Ser Val Leu Pro Val Cys Leu Ala Thr Pro Glu Val
165 170 175
Asn Leu Thr Ser Ala Asn Cys Trp Ala Thr Gly Trp Gly Leu Val Ser
180 185 190
Lys Gln Gly Glu Thr Ser Asp Glu Leu Gln Glu Met Gln Leu Pro Leu
195 200 205
Ile Leu Glu Pro Trp Cys His Leu Leu Tyr Gly His Met Ser Tyr Ile
210 215 220
Met Pro Asp Met Leu Cys Ala Gly Asp Ile Leu Asn Ala Lys Thr Val
225 230 235 240
Cys Glu Gly Asp Ser Gly Gly Pro Leu Val Cys Glu Phe Asn Arg Ser
245 250 255
Trp Leu Gln Ile Gly Ile Val Ser Trp Gly Arg Gly Cys Ser Asn Pro
260 265 270
Leu Tyr Pro Gly Val Tyr Ala Ser Val Ser Tyr Phe Ser Lys Trp Ile
275 280 285
Cys Asp Asn Ile Glu Ile Thr Pro Thr Pro Ala Gln Pro Ala Pro Ala
290 295 300
Leu Ser Pro Ala Leu Gly Pro Thr Leu Ser Val Leu Met Ala Met Leu
305 310 315 320
Ala Gly Trp Ser Val Leu
325
<210>267
<211>1695
<212>DNA
<213>Homo sapiens
<220>
<223>Mername-AA180 peptidase
<400>267
atgcttataa gcaggaacaa gctgatttta ctactaggaa tagtcttttt tgaacgaggt 60
aaatctgcaa ctctttcgct ccccaaagct cccagttgtg ggcagagtct ggttaaggta 120
cagccttgga attattttaa cattttcagt cgcattcttg gaggaagcca agtggagaag 180
ggttcctatc cctggcaggt atctctgaaa caaaggcaga agcatatttg tggaggaagc 240
atcgtctcac cacagtgggt gatcacggcg gctcactgca ttgcaaacag aaacattgtg 300
tctactttga atgttactgc tggagagtat gacttaagcc agacagaccc aggagagcaa 360
actctcacta ttgaaactgt catcatacat ccacatttct ccaccaagaa accaatggac 420
tatgatattg cccttttgaa gatggctgga gccttccaat ttggccactt tgtggggccc 480
atatgtcttc cagagctgcg ggagcaattt gaggctggtt ttatttgtac aactgcaggc 540
tggggccgct taactgaagg tggcgtcctc tcacaagtct tgcaggaagt gaatctgcct 600
attttgacct gggaagagtg tgtggcagct ctgttaacac taaagaggcc catcagtggg 660
aagacctttc tttgcacagg ttttcctgat ggagggagag acgcatgtca gggagattca 720
ggaggttcac tcatgtgccg gaataagaaa ggggcctgga ctctggctgg tgtgacttcc 780
tggggtttgg gctgtggtcg aggctggaga aacaatgtga ggaaaagtga tcaaggatcc 840
cctgggatct tcacagacat tagtaaagtg cttccctgga tccacgaaca catccaaact 900
ggtaatcgga gaaagagctc cagagcctgg tgcagtgagc aggatgtcat agtcagcggg 960
gctgagggga agctgcactt cccagaaagc ctccacctat attatgagag caagcaacgg 1020
tgtgtctgga ccctgctggt accagaggaa atgcatgtgt tgctcagttt ttcccaccta 1080
gatgttgagt cttgtcacca cagttacctg tcaatgtatt ctttagaaga cagacccatt 1140
ggaaaatttt gtggagaaag cctcccttca tccattctta ttggctctaa ttctctaagg 1200
ctgaaattcg tctctgatgc cacagattat gcagctgggt ttaatcttac ctataaagct 1260
cttaaaccaa actacattcc tgattcaggt tgcagttact taactgtcct ttttgaagaa 1320
ggtctcatac agagtctaaa ctatcctgaa aactacagtg acaaggctaa ctgtgactgg 1380
atttttcaag cctccaaaca tcacctaatt aagctttcat ttcagagtct ggaaatagaa 1440
gaaagtggag actgcacttc cgactatgtg acagtgcaca gcgatgtaga aaggaagaag 1500
gaaatagctc ggctgtgtgg ctatgatgtc cccacccctg tgctgagccc ctccagcatc 1560
atgctcatca gcttccaatc agatgaaaac gggacctgca ggggctttca ggctacagtc 1620
tccttcattc ctaaagcagg taagaagata gagttaccaa ctctttggtt cccagttcta 1680
atccttgtga tgtga 1695
<210>268
<211>564
<212>PRT
<213>Homo sapiens
<220>
<223>Mername-AA180 peptidase
<400>268
Met Leu Ile Ser Arg Asn Lys Leu Ile Leu Leu Leu Gly Ile Val Phe
1 5 10 15
Phe Glu Arg Gly Lys Ser Ala Thr Leu Ser Leu Pro Lys Ala Pro Ser
20 25 30
Cys Gly Gln Ser Leu Val Lys Val Gln Pro Trp Asn Tyr Phe Asn Ile
35 40 45
Phe Ser Arg Ile Leu Gly Gly Ser Gln Val Glu Lys Gly Ser Tyr Pro
50 55 60
Trp Gln Val Ser Leu Lys Gln Arg Gln Lys His Ile Cys Gly Gly Ser
65 70 75 80
Ile Val Ser Pro Gln Trp Val Ile Thr Ala Ala His Cys Ile Ala Asn
85 90 95
Arg Asn Ile Val Ser Thr Leu Asn Val Thr Ala Gly Glu Tyr Asp Leu
100 105 110
Ser Gln Thr Asp Pro Gly Glu Gln Thr Leu Thr Ile Glu Thr Val Ile
115 120 125
Ile His Pro His Phe Ser Thr Lys Lys Pro Met Asp Tyr Asp Ile Ala
130 135 140
Leu Leu Lys Met Ala Gly Ala Phe Gln Phe Gly His Phe Val Gly Pro
145 150 155 160
Ile Cys Leu Pro Glu Leu Arg Glu Gln Phe Glu Ala Gly Phe Ile Cys
165 170 175
Thr Thr Ala Gly Trp Gly Arg Leu Thr Glu Gly Gly Val Leu Ser Gln
180 185 190
Val Leu Gln Glu Val Asn Leu Pro Ile Leu Thr Trp Glu Glu Cys Val
195 200 205
Ala Ala Leu Leu Thr Leu Lys Arg Pro Ile Ser Gly Lys Thr Phe Leu
210 215 220
Cys Thr Gly Phe Pro Asp Gly Gly Arg Asp Ala Cys Gln Gly Asp Ser
225 230 235 240
Gly Gly Ser Leu Met Cys Arg Asn Lys Lys Gly Ala Trp Thr Leu Ala
245 250 255
Gly Val Thr Ser Trp Gly Leu Gly Cys Gly Arg Gly Trp Arg Asn Asn
260 265 270
Val Arg Lys Ser Asp Gln Gly Ser Pro Gly Ile Phe Thr Asp Ile Ser
275 280 285
Lys Val Leu Pro Trp Ile His Glu His Ile Gln Thr Gly Asn Arg Arg
290 295 300
Lys Ser Ser Arg Ala Trp Cys Ser Glu Gln Asp Val Ile Val Ser Gly
305 310 315 320
Ala Glu Gly Lys Leu His Phe Pro Glu Ser Leu His Leu Tyr Tyr Glu
325 330 335
Ser Lys Gln Arg Cys Val Trp Thr Leu Leu Val Pro Glu Glu Met His
340 345 350
Val Leu Leu Ser Phe Ser His Leu Asp Val Glu Ser Cys His His Ser
355 360 365
Tyr Leu Ser Met Tyr Ser Leu Glu Asp Arg Pro Ile Gly Lys Phe Cys
370 375 380
Gly Glu Ser Leu Pro Ser Ser Ile Leu Ile Gly Ser Asn Ser Leu Arg
385 390 395 400
Leu Lys Phe Val Ser Asp Ala Thr Asp Tyr Ala Ala Gly Phe Asn Leu
405 410 415
Thr Tyr Lys Ala Leu Lys Pro Asn Tyr Ile Pro Asp Ser Gly Cys Ser
420 425 430
Tyr Leu Thr Val Leu Phe Glu Glu Gly Leu Ile Gln Ser Leu Asn Tyr
435 440 445
Pro Glu Asn Tyr Ser Asp Lys Ala Asn Cys Asp Trp Ile Phe Gln Ala
450 455 460
Ser Lys His His Leu Ile Lys Leu Ser Phe Gln Ser Leu Glu Ile Glu
465 470 475 480
Glu Ser Gly Asp Cys Thr Ser Asp Tyr Val Thr Val His Ser Asp Val
485 490 495
Glu Arg Lys Lys Glu Ile Ala Arg Leu Cys Gly Tyr Asp Val Pro Thr
500 505 510
Pro Val Leu Ser Pro Ser Ser Ile Met Leu Ile Ser Phe Gln Ser Asp
515 520 525
Glu Asn Gly Thr Cys Arg Gly Phe Gln Ala Thr Val Ser Phe Ile Pro
530 535 540
Lys Ala Gly Lys Lys Ile Glu Leu Pro Thr Leu Trp Phe Pro Val Leu
545 550 555 560
Ile Leu Val Met
<210>269
<211>3420
<212>DNA
<213>Homo sapiens
<220>
<223>Mername-AA182 peptidase
<400>269
atgggcctgc tggccagtgc tggtttgttg ctgttgctgg tcatcggcca ccccagaagc 60
ctaggactga agtgtggaat tcgcatggtc aacatgaaaa gtaaggaacc tgccgtggga 120
tctagattct tctctagaat tagtagttgg agaaattcaa cagtgactgg acatccatgg 180
caggtctccc taaaatcaga tgagcaccac ttctgtggag gaagcttgat tcaagaagat 240
cgggttgtta cagcagcaca ctgcctggac agcctcagtg agaagcagct gaagaatata 300
actgtgactt ctggggagta cagcctcttt cagaaggata agcaagaaca gaatattcct 360
gtctcaaaaa ttattaccca tcctgaatac aacagccgtg aatatatgag tcctgatatt 420
gcactgctgt atctaaaaca caaagtcaag tttggaaatg ctgttcagcc aatctgtctt 480
cctgacagcg atgataaagt tgaaccagga attctttgct tatccagtgg atggggcaag 540
atttccaaaa catcagaata ttcaaatgtc ctacaagaaa tggaacttcc catcatggat 600
gacagagcgt gtaatactgt gctcaagagc atgaacctcc ctcccctggg aaggaccatg 660
ctgtgtgctg gcttccctga ttggggaatg gacgcctgcc agggggactc tggaggacca 720
ctggtttgta gaagaggtgg tggaatctgg attcttgctg ggataacttc ctgggtagct 780
ggttgtgctg gaggttcagt tcccgtaaga aacaaccatg tgaaggcatc acttggcatt 840
ttctccaaag tgtctgagtt gatggatttt atcactcaaa acctgttcac aggtttggat 900
cggggccaac ccctctcaaa agtgggctca aggtatataa caaaggccct gagttctgtc 960
caagaagtga atggaagcca gagaggaaag ggtattttag acatggaaaa gcaagttgga 1020
tgtgatcatg actatgtatc tttacgatca agcagtggag tgctttttaa tcagagatct 1080
ctcatggaag acgatggaaa acaaaacaaa cgtgtctgtg gaaaaatatt gccttcacca 1140
ttgctggcag agaccagtga ggccatggtt ccatttgttt ctgatacaga agacagtggc 1200
agtggctttg agcttaccgt tactgctgta cagaagtcag aagcagggtc aggttgtggg 1260
agtctggcta tattggtaga agaagggaca aatcactctg ccaagtatcc tgatttgtat 1320
cccagtaaca caaggtgtca ttggttcatt tgtgctccag agaagcacat tataaagttg 1380
acatttgagg actttgctgt caaatttagt ccaaactgta tttatgatgc tgttgtgatt 1440
tacggtgatt ctgaagaaaa gcacaagtta gctaaacttt gtggaatgtt gaccatcact 1500
tcaatattca gttctagtaa catgacggtg atatacttta aaagtgatgg taaaaatcgt 1560
ttacaaggct tcaaggccag atttaccatt ttgccctcag agtctttaaa caaatttgaa 1620
ccaaagttac ctccccaaaa caatcctgta tctaccgtaa aagctattct gcatgatgtc 1680
tgtggcatcc ctccatttag tccccagtgg ctttccagaa gaatcgcagg aggggaagaa 1740
gcctgccccc actgttggcc atggcaggtg ggtctgaggt ttctaggcga ttaccaatgt 1800
ggaggtgcca tcatcaaccc agtgtggatt ctgaccgcag cccactgtgt gcaattgaag 1860
aataatccac tctcctggac tattattgct ggggaccatg acagaaacct gaaggaatca 1920
acagagcagg tgagaagggc caaacacata atagtgcatg aagactttaa cacactaagt 1980
tatgactctg acattgccct aatacaacta agctctcctc tggagtacaa ctcggtggtg 2040
aggccagtat gtctcccaca cagcgcagag cctctatttt cctcggagat ctgtgctgtg 2100
accggatggg gaagcatcag tgcagatggt ggcctagcaa gtcgcctaca gcagattcaa 2160
gtgcatgtgt tagaaagaga ggtctgtgaa cacacttact attctgccca tccaggaggg 2220
atcacagaga agatgatctg tgctggcttt gcagcatctg gagagaaaga tttctgccag 2280
ggagactctg gtgggccact agtatgtaga catgaaaatg gtccctttgt cctctatggc 2340
attgtcagct ggggagctgg ctgtgtccag ccatggaagc cgggtgtatt tgccagagtg 2400
atgatcttct tggactggat ccaatcaaaa atcaatggtc ctgcttcact tcagacaaat 2460
aataaatgca aaaccttaaa acaacaattg ccaccaccca caccttcacc agacagtgca 2520
tcttggccag gttgttgctc tgaagcagag ctagaaaagc ctagaggctt ttttcccaca 2580
ccacggtatc tactggatta tagaggaaga ctggaatgtt cttgggtgct cagagtttca 2640
gcaagcagta tggcaaaatt taccattgag tatctgtcac tcctggggtc tcctgtgtgt 2700
caagactcag ttctaattat ttatgaagaa agacacagta agagaaagac ggcaggtgga 2760
ttacatggaa gaagacttta ctcaatgact ttcatgagtc ctggaccgct ggtgagggtg 2820
acattccatg cccttgtacg aggtgcattt ggtataagct atattgactt gaaagtccta 2880
ggtccaaagg acagtaaaat aaccagactt tcccaaagtt caaacagaga gcacttggtc 2940
ccttgtgagg atgttcttct gaccaagcca gaagggatca tgcagatccc aagaaattct 3000
cacagaacta ctatggggtg ccaatggaga ttagtagccc ctttaaatca catcattcag 3060
cttaatatta ttaacttccc gatgaagcca acaacttttg tctgtcatgg tcatctgcgt 3120
gtttacgaag gatttggacc aggaaaaaaa ttaatagctt cattcgctgg aactctggca 3180
atgatactga caaaagacat attaaaaaga gaaaaactga attttattaa cacatatatc 3240
atgcatatat gggaaaattc agtgtatgat aatgtaagaa gtgttggaaa aagaaaacag 3300
aagaaatttg catcaaattt aagttatagt atggaggctg agaagtccag gatccaggtg 3360
ccagcagatt tggtgcctgc taagggctca ctttctggtt cttaggcaca gcccattctc 3420
<210>270
<211>1134
<212>PRT
<213>Homo sapiens
<220>
<223>Mername-AA182 peptidase
<400>270
Met Gly Leu Leu Ala Ser Ala Gly Leu Leu Leu Leu Leu Val Ile Gly
1 5 10 15
His Pro Arg Ser Leu Gly Leu Lys Cys Gly Ile Arg Met Val Asn Met
20 25 30
Lys Ser Lys Glu Pro Ala Val Gly Ser Arg Phe Phe Ser Arg Ile Ser
35 40 45
Ser Trp Arg Asn Ser Thr Val Thr Gly His Pro Trp Gln Val Ser Leu
50 55 60
Lys Ser Asp Glu His His Phe Cys Gly Gly Ser Leu Ile Gln Glu Asp
65 70 75 80
Arg Val Val Thr Ala Ala His Cys Leu Asp Ser Leu Ser Glu Lys Gln
85 90 95
Leu Lys Asn Ile Thr Val Thr Ser Gly Glu Tyr Ser Leu Phe Gln Lys
100 105 110
Asp Lys Gln Glu Gln Asn Ile Pro Val Ser Lys Ile Ile Thr His Pro
115 120 125
Glu Tyr Asn Ser Arg Glu Tyr Met Ser Pro Asp Ile Ala Leu Leu Tyr
130 135 140
Leu Lys His Lys Val Lys Phe Gly Asn Ala Val Gln Pro Ile Cys Leu
145 150 155 160
Pro Asp Ser Asp Asp Lys Val Glu Pro Gly Ile Leu Cys Leu Ser Ser
165 170 175
Gly Trp Gly Lys Ile Ser Lys Thr Ser Glu Tyr Ser Asn Val Leu Gln
180 185 190
Glu Met Glu Leu Pro Ile Met Asp Asp Arg Ala Cys Asn Thr Val Leu
195 200 205
Lys Ser Met Asn Leu Pro Pro Leu Gly Arg Thr Met Leu Cys Ala Gly
210 215 220
Phe Pro Asp Trp Gly Met Asp Ala Cys Gln Gly Asp Ser Gly Gly Pro
225 230 235 240
Leu Val Cys Arg Arg Gly Gly Gly Ile Trp Ile Leu Ala Gly Ile Thr
245 250 255
Ser Trp Val Ala Gly Cys Ala Gly Gly Ser Val Pro Val Arg Asn Asn
260 265 270
His Val Lys Ala Ser Leu Gly Ile Phe Ser Lys Val Ser Glu Leu Met
275 280 285
Asp Phe Ile Thr Gln Asn Leu Phe Thr Gly Leu Asp Arg Gly Gln Pro
290 295 300
Leu Ser Lys Val Gly Ser Arg Tyr Ile Thr Lys Ala Leu Ser Ser Val
305 310 315 320
Gln Glu Val Asn Gly Ser Gln Arg Gly Lys Gly Ile Leu Asp Met Glu
325 330 335
Lys Gln Val Gly Cys Asp His Asp Tyr Val Ser Leu Arg Ser Ser Ser
340 345 350
Gly Val Leu Phe Asn Gln Arg Ser Leu Met Glu Asp Asp Gly Lys Gln
355 360 365
Asn Lys Arg Val Cys Gly Lys Ile Leu Pro Ser Pro Leu Leu Ala Glu
370 375 380
Thr Ser Glu Ala Met Val Pro Phe Val Ser Asp Thr Glu Asp Ser Gly
385 390 395 400
Ser Gly Phe Glu Leu Thr Val Thr Ala Val Gln Lys Ser Glu Ala Gly
405 410 415
Ser Gly Cys Gly Ser Leu Ala Ile Leu Val Glu Glu Gly Thr Asn His
420 425 430
Ser Ala Lys Tyr Pro Asp Leu Tyr Pro Ser Asn Thr Arg Cys His Trp
435 440 445
Phe Ile Cys Ala Pro Glu Lys His Ile Ile Lys Leu Thr Phe Glu Asp
450 455 460
Phe Ala Val Lys Phe Ser Pro Asn Cys Ile Tyr Asp Ala Val Val Ile
465 470 475 480
Tyr Gly Asp Ser Glu Glu Lys His Lys Leu Ala Lys Leu Cys Gly Met
485 490 495
Leu Thr Ile Thr Ser Ile Phe Ser Ser Ser Asn Met Thr Val Ile Tyr
500 505 510
Phe Lys Ser Asp Gly Lys Asn Arg Leu Gln Gly Phe Lys Ala Arg Phe
515 520 525
Thr Ile Leu Pro Ser Glu Ser Leu Asn Lys Phe Glu Pro Lys Leu Pro
530 535 540
Pro Gln Asn Asn Pro Val Ser Thr Val Lys Ala Ile Leu His Asp Val
545 550 555 560
Cys Gly Ile Pro Pro Phe Ser Pro Gln Trp Leu Ser Arg Arg Ile Ala
565 570 575
Gly Gly Glu Glu Ala Cys Pro His Cys Trp Pro Trp Gln Val Gly Leu
580 585 590
Arg Phe Leu Gly Asp Tyr Gln Cys Gly Gly Ala Ile Ile Asn Pro Val
595 600 605
Trp Ile Leu Thr Ala Ala His Cys Val Gln Leu Lys Asn Asn Pro Leu
610 615 620
Ser Trp Thr Ile Ile Ala Gly Asp His Asp Arg Asn Leu Lys Glu Ser
625 630 635 640
Thr Glu Gln Val Arg Arg Ala Lys His Ile Ile Val His Glu Asp Phe
645 650 655
Asn Thr Leu Ser Tyr Asp Ser Asp Ile Ala Leu Ile Gln Leu Ser Ser
660 665 670
Pro Leu Glu Tyr Asn Ser Val Val Arg Pro Val Cys Leu Pro His Ser
675 680 685
Ala Glu Pro Leu Phe Ser Ser Glu Ile Cys Ala Val Thr Gly Trp Gly
690 695 700
Ser Ile Ser Ala Asp Gly Gly Leu Ala Ser Arg Leu Gln Gln Ile Gln
705 710 715 720
Val His Val Leu Glu Arg Glu Val Cys Glu His Thr Tyr Tyr Ser Ala
725 730 735
His Pro Gly Gly Ile Thr Glu Lys Met Ile Cys Ala Gly Phe Ala Ala
740 745 750
Ser Gly Glu Lys Asp Phe Cys Gln Gly Asp Ser Gly Gly Pro Leu Val
755 760 765
Cys Arg His Glu Asn Gly Pro Phe Val Leu Tyr Gly Ile Val Ser Trp
770 775 780
Gly Ala Gly Cys Val Gln Pro Trp Lys Pro Gly Val Phe Ala Arg Val
785 790 795 800
Met Ile Phe Leu Asp Trp Ile Gln Ser Lys Ile Asn Gly Pro Ala Ser
805 810 815
Leu Gln Thr Asn Asn Lys Cys Lys Thr Leu Lys Gln Gln Leu Pro Pro
820 825 830
Pro Thr Pro Ser Pro Asp Ser Ala Ser Trp Pro Gly Cys Cys Ser Glu
835 840 845
Ala Glu Leu Glu Lys Pro Arg Gly Phe Phe Pro Thr Pro Arg Tyr Leu
850 855 860
Leu Asp Tyr Arg Gly Arg Leu Glu Cys Ser Trp Val Leu Arg Val Ser
865 870 875 880
Ala Ser Ser Met Ala Lys Phe Thr Ile Glu Tyr Leu Ser Leu Leu Gly
885 890 895
Ser Pro Val Cys Gln Asp Ser Val Leu Ile Ile Tyr Glu Glu Arg His
900 905 910
Ser Lys Arg Lys Thr Ala Gly Gly Leu His Gly Arg Arg Leu Tyr Ser
915 920 925
Met Thr Phe Met Ser Pro Gly Pro Leu Val Arg Val Thr Phe His Ala
930 935 940
Leu Val Arg Gly Ala Phe Gly Ile Ser Tyr Ile Asp Leu Lys Val Leu
945 950 955 960
Gly Pro Lys Asp Ser Lys Ile Thr Arg Leu Ser Gln Ser Ser Asn Arg
965 970 975
Glu His Leu Val Pro Cys Glu Asp Val Leu Leu Thr Lys Pro Glu Gly
980 985 990
Ile Met Gln Ile Pro Arg Asn Ser His Arg Thr Thr Met Gly Cys Gln
995 1000 1005
Trp Arg Leu Val Ala Pro Leu Asn His Ile Ile Gln Leu Asn Ile Ile
1010 1015 1020
Asn Phe Pro Met Lys Pro Thr Thr Phe Val Cys His Gly His Leu Arg
1025 1030 1035 1040
Val Tyr Glu Gly Phe Gly Pro Gly Lys Lys Leu Ile Ala Ser Phe Ala
1045 1050 1055
Gly Thr Leu Ala Met Ile Leu Thr Lys Asp Ile Leu Lys Arg Glu Lys
1060 1065 1070
Leu Asn Phe Ile Asn Thr Tyr Ile Met His Ile Trp Glu Asn Ser Val
1075 1080 1085
Tyr Asp Asn Val Arg Ser Val Gly Lys Arg Lys Gln Lys Lys Phe Ala
1090 1095 1100
Ser Asn Leu Ser Tyr Ser Met Glu Ala Glu Lys Ser Arg Ile Gln Val
1105 1110 1115 1120
Pro Ala Asp Leu Val Pro Ala Lys Gly Ser Leu Ser Gly Ser
1125 1130
<210>271
<211>2810
<212>DNA
<213>Homo sapiens
<220>
<223>Mername-AA122 peptidase
<400>271
acttggctct gcctccgctg actctagcgc catggcccgg cacctgctcc tcccccttgt 60
gatgcttgtc atcagtccca tcccaggagc cttccaggac tcagctctca gtcctaccca 120
ggaagaacct gaagatctgg actgcgggcg ccctgagccc tcggcccgca tcgtgggggg 180
gctcaaacgc gcagccgggc acctggcctt ggcaagtgag cctgcaccat ggaggtggcc 240
acatctgcgg gggctccctc atcgccccct cctgggtcct ctccgctgct cactgtttca 300
tgacgaatgg gacgctggag cccgcggccg agtggtcggt actgctgggc gtgcactccc 360
aggacgggcc cctggacggc gcgcacaccc gcgcagtggc cgccatcgtg gtgccggcca 420
actacagcca agtggagctg ggcgccgacc tggccctgct gcgcctggcc tcacccgcca 480
gcctgggccc cgccgtgtgg cctgtctgcc tgccccgcgc ctcacaccgc ttcgtgcacg 540
gcaccgcctg ctgggccacc ggctggggag acgtccagga ggcagatcct ctgcctctcc 600
cctgggtgct acaggaagtg gagctaaggc tgctgggcga ggccacctgt caatgtctct 660
acagccagcc cggtcccttc aacctcactc tccagatatt gccagggatg ctgtgtgctg 720
gctacccaga gggccgcagg gacacctgcc agggcgactc tggggggccc ctggtctgtg 780
aggaaggcgg ccgctggttc caggcaggaa tcaccagctt tggctttggc tgtggacgga 840
gaaaccgccc tggagttttc actgctgtgg ctacctatga ggcatggata cgggagcagg 900
tgatgggttc agagcctggg cctgcctttc ccacccagcc ccagaagacc cagtcagatc 960
cccaggagcc cagggaggag aactgcacca ttgccctgcc tgagtgcggg aaggccccgc 1020
ggccaggggc ctggccctgg gaggcccagg tgatggtgcc aggatccaga ccctgccatg 1080
gggcgctggt gtctgaaagc tgggtcttgg cacctgccag ctgctttctg gacccgaaca 1140
gctccgacag cccaccccgc gacctcgacg cctggcgcgt gctgctgccc tcgcgcccgc 1200
gcgcggagcg ggtggcgcgc ctggtgcagc acgagaacgc ttcgtgggac aacgcctcgg 1260
acctggcgct gctgcagctg cgcacgcccg tgaacctgag tgcggcttcg cggcccgtgt 1320
gcctacccca cccggaacac tacttcctgc ccgggagccg ctgccgcctg gcccgctggg 1380
gccgcgggga acccgcgctt ggcccaggcg cgctgctgga ggcggagctg ttaggcggct 1440
ggtggtgcca ctgcctgtac ggccgccagg gggcggcagt accgctgccc ggagacccgc 1500
cgcacgcgct ctgccctgcc taccaggaaa aggaggaggt gggcagctgc tggaatgact 1560
cgcgttggag ccttttgtgc caggaggagg ggacctggtt tctggctgga atcagagact 1620
ttcccagtgg ctgtctacgt ccccgagcct tcttccctct gcagactcat ggcccatgga 1680
tcagccatgt gactcgggga gcctacctgg aggaccagct agcctgggac tggggccctg 1740
atggggagga gactgagaca cagacttgtc ccccacacac agagcatggt gcctgtggcc 1800
tgcggctgga ggctgctcca gtgggggtcc tgtggccctg gctggcagag gtgcatgtgg 1860
ctggtgatcg agtctgcact gggatcctcc tggccccagg ctgggtcctg gcagccactc 1920
actgtgtcct caggccaggc tctacaacag tgccttacat tgaagtgtat ctgggccggg 1980
caggggccag ctccctccca cagggccacc aggtatcccg cttggtcatc agcatccggc 2040
tgccccagca cctgggactc aggccccccc tggccctcct ggagctgagc tcccgggtgg 2100
agccctcccc atcagccctg cccatctgtc tccacccggc gggtatcccc ccgggggcca 2160
gctgctgggt gttgggctgg aaagaacccc aggaccgagt ccctgtggct gctgctgtct 2220
ccatcttgac acaacgaatc tgtgactgcc tctatcaggg catcctgccc cctggaaccc 2280
tctgtgtcct gtatgcagag gggcaggaga acaggtgtga gatgacctca gcaccgcccc 2340
tcctgtgcca gatgacggaa gggtcctgga tcctcgtggg catggctgtt caagggagcc 2400
gggagctgtt tgctgccatt ggtcctgaag aggcctggat ctcccagaca gtgggagagg 2460
ccaacctcct gccccccagt ggctccccac actggcccac tggaggcagc aatctctgcc 2520
ccccagaact ggccaaggcc tcgggatccc cgcatgcagt ctacttcctg ctcctgctga 2580
ctctcctgat ccagagctga ggggctaggg tcccagcacc acttccccct tctccaccct 2640
ctacttcccg cccagtgggg ctggaatgtg gcccagccgg ctggaacctc aagggcccca 2700
cccaccgaga ttgcagcggc tctggctaat tgggcctctg tgcccgggct attttgaacc 2760
caggaatcct tgggggtggt gggaggagcg gacaataaag gtgtaaacac 2810
<210>272
<211>766
<212>PRT
<213>Homo sapiens
<220>
<223>Mername-AA122 peptidase
<400>272
Met Thr Asn Gly Thr Leu Glu Pro Ala Ala Glu Trp Ser Val Leu Leu
1 5 10 15
Gly Val His Ser Gln Asp Gly Pro Leu Asp Gly Ala His Thr Arg Ala
20 25 30
Val Ala Ala Ile Val Val Pro Ala Asn Tyr Ser Gln Val Glu Leu Gly
35 40 45
Ala Asp Leu Ala Leu Leu Arg Leu Ala Ser Pro Ala Ser Leu Gly Pro
50 55 60
Ala Val Trp Pro Val Cys Leu Pro Arg Ala Ser His Arg Phe Val His
65 70 75 80
Gly Thr Ala Cys Trp Ala Thr Gly Trp Gly Asp Val Gln Glu Ala Asp
85 90 95
Pro Leu Pro Leu Pro Trp Val Leu Gln Glu Val Glu Leu Arg Leu Leu
100 105 110
Gly Glu Ala Thr Cys Gln Cys Leu Tyr Ser Gln Pro Gly Pro Phe Asn
115 120 125
Leu Thr Leu Gln Ile Leu Pro Gly Met Leu Cys Ala Gly Tyr Pro Glu
130 135 140
Gly Arg Arg Asp Thr Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys
145 150 155 160
Glu Glu Gly Gly Arg Trp Phe Gln Ala Gly Ile Thr Ser Phe Gly Phe
165 170 175
Gly Cys Gly Arg Arg Asn Arg Pro Gly Val Phe Thr Ala Val Ala Thr
180 185 190
Tyr Glu Ala Trp Ile Arg Glu Gln Val Met Gly Ser Glu Pro Gly Pro
195 200 205
Ala Phe Pro Thr Gln Pro Gln Lys Thr Gln Ser Asp Pro Gln Glu Pro
210 215 220
Arg Glu Glu Asn Cys Thr Ile Ala Leu Pro Glu Cys Gly Lys Ala Pro
225 230 235 240
Arg Pro Gly Ala Trp Pro Trp Glu Ala Gln Val Met Val Pro Gly Ser
245 250 255
Arg Pro Cys His Gly Ala Leu Val Ser Glu Ser Trp Val Leu Ala Pro
260 265 270
Ala Ser Cys Phe Leu Asp Pro Asn Ser Ser Asp Ser Pro Pro Arg Asp
275 280 285
Leu Asp Ala Trp Arg Val Leu Leu Pro Ser Arg Pro Arg Ala Glu Arg
290 295 300
Val Ala Arg Leu Val Gln His Glu Asn Ala Ser Trp Asp Asn Ala Ser
305 310 315 320
Asp Leu Ala Leu Leu Gln Leu Arg Thr Pro Val Asn Leu Ser Ala Ala
325 330 335
Ser Arg Pro Val Cys Leu Pro His Pro Glu His Tyr Phe Leu Pro Gly
340 345 350
Ser Arg Cys Arg Leu Ala Arg Trp Gly Arg Gly Glu Pro Ala Leu Gly
355 360 365
Pro Gly Ala Leu Leu Glu Ala Glu Leu Leu Gly Gly Trp Trp Cys His
370 375 380
Cys Leu Tyr Gly Arg Gln Gly Ala Ala Val Pro Leu Pro Gly Asp Pro
385 390 395 400
Pro His Ala Leu Cys Pro Ala Tyr Gln Glu Lys Glu Glu Val Gly Ser
405 410 415
Cys Trp Asn Asp Ser Arg Trp Ser Leu Leu Cys Gln Glu Glu Gly Thr
420 425 430
Trp Phe Leu Ala Gly Ile Arg Asp Phe Pro Ser Gly Cys Leu Arg Pro
435 440 445
Arg Ala Phe Phe Pro Leu Gln Thr His Gly Pro Trp Ile Ser His Val
450 455 460
Thr Arg Gly Ala Tyr Leu Glu Asp Gln Leu Ala Trp Asp Trp Gly Pro
465 470 475 480
Asp Gly Glu Glu Thr Glu Thr Gln Thr Cys Pro Pro His Thr Glu His
485 490 495
Gly Ala Cys Gly Leu Arg Leu Glu Ala Ala Pro Val Gly Val Leu Trp
500 505 510
Pro Trp Leu Ala Glu Val His Val Ala Gly Asp Arg Val Cys Thr Gly
515 520 525
Ile Leu Leu Ala Pro Gly Trp Val Leu Ala Ala Thr His Cys Val Leu
530 535 540
Arg Pro Gly Ser Thr Thr Val Pro Tyr Ile Glu Val Tyr Leu Gly Arg
545 550 555 560
Ala Gly Ala Ser Ser Leu Pro Gln Gly His Gln Val Ser Arg Leu Val
565 570 575
Ile Ser Ile Arg Leu Pro Gln His Leu Gly Leu Arg Pro Pro Leu Ala
580 585 590
Leu Leu Glu Leu Ser Ser Arg Val Glu Pro Ser Pro Ser Ala Leu Pro
595 600 605
Ile Cys Leu His Pro Ala Gly Ile Pro Pro Gly Ala Ser Cys Trp Val
610 615 620
Leu Gly Trp Lys Glu Pro Gln Asp Arg Val Pro Val Ala Ala Ala Val
625 630 635 640
Ser Ile Leu Thr Gln Arg Ile Cys Asp Cys Leu Tyr Gln Gly Ile Leu
645 650 655
Pro Pro Gly Thr Leu Cys Val Leu Tyr Ala Glu Gly Gln Glu Asn Arg
660 665 670
Cys Glu Met Thr Ser Ala Pro Pro Leu Leu Cys Gln Met Thr Glu Gly
675 680 685
Ser Trp Ile Leu Val Gly Met Ala Val Gln Gly Ser Arg Glu Leu Phe
690 695 700
Ala Ala Ile Gly Pro Glu Glu Ala Trp Ile Ser Gln Thr Val Gly Glu
705 710 715 720
Ala Asn Leu Leu Pro Pro Ser Gly Ser Pro His Trp Pro Thr Gly Gly
725 730 735
Ser Asn Leu Cys Pro Pro Glu Leu Ala Lys Ala Ser Gly Ser Pro His
740 745 750
Ala Val Tyr Phe Leu Leu Leu Leu Thr Leu Leu Ile Gln Ser
755 760 765
<210>273
<211>1575
<212>DNA
<213>Homo sapiens
<220>
<223>Cathepsin L
<400>273
agaaccgcga cctccgcaac cttgagcggc atccgtggag tgcgcctgca gctacgaccg 60
cagcaggaaa gcgccgccgg ccaggcccag ctgtggccgg acagggactg gaagagagga 120
cgcggtcgag taggtgtgca ccagccctgg caacgagagc gtctaccccg aactctgctg 180
gccttgaggt ggggaagccg gggagggcag ttgaggaccc cgcggaggcg cgtgactggt 240
tgagcgggca ggccagcctc cgagccgggt ggacacaggt tttaaaacat gaatcctaca 300
ctcatccttg ctgccttttg cctgggaatt gcctcagcta ctctaacatt tgatcacagt 360
ttagaggcac agtggaccaa gtggaaggcg atgcacaaca gattatacgg catgaatgaa 420
gaaggatgga ggagagcagt gtgggagaag aacatgaaga tgattgaact gcacaatcag 480
gaatacaggg aagggaaaca cagcttcaca atggccatga acgcctttgg agacatgacc 540
agtgaagaat tcaggcaggt gatgaatggc tttcaaaacc gtaagcccag gaaggggaaa 600
gtgttccagg aacctctgtt ttatgaggcc cccagatctg tggattggag agagaaaggc 660
tacgtgactc ctgtgaagaa tcagggtcag tgtggttctt gttgggcttt tagtgctact 720
ggtgctcttg aaggacagat gttccggaaa actgggaggc ttatctcact gagtgagcag 780
aatctggtag actgctctgg gcctcaaggc aatgaaggct gcaatggtgg cctaatggat 840
tatgctttcc agtatgttca ggataatgga ggcctggact ctgaggaatc ctatccatat 900
gaggcaacag aagaatcctg taagtacaat cccaagtatt ctgttgctaa tgacaccggc 960
tttgtggaca tccctaagca ggagaaggcc ctgatgaagg cagttgcaac tgtggggccc 1020
atttctgttg ctattgatgc aggtcatgag tccttcctgt tctataaaga aggcatttat 1080
tttgagccag actgtagcag tgaagacatg gatcatggtg tgctggtggt tggctacgga 1140
tttgaaagca cagaatcaga taacaataaa tattggctgg tgaagaacag ctggggtgaa 1200
gaatggggca tgggtggcta cgtaaagatg gccaaagacc ggagaaacca ttgtggaatt 1260
gcctcagcag ccagctaccc cactgtgtga gctggtggac ggtgatgagg aaggacttga 1320
ctggggatgg cgcatgcatg ggaggaattc atcttcagtc taccagcccc cgctgtgtcg 1380
gatacacact cgaatcattg aagatccgag tgtgatttga attctgtgat attttcacac 1440
tggtaaatgt tacctctatt ttaattactg ctataaatag gtttatatta ttgattcact 1500
tactgacttt gcattttcgt ttttaaaagg atgtataaat ttttacctgt ttaaataaaa 1560
tttaatttca aatgt 1575
<210>274
<211>333
<212>PRT
<213>Homo sapiens
<220>
<223>Cathepsin L
<400>274
Met Asn Pro Thr Leu Ile Leu Ala Ala Phe Cys Leu Gly Ile Ala Ser
1 5 10 15
Ala Thr Leu Thr Phe Asp His Ser Leu Glu Ala Gln Trp Thr Lys Trp
20 25 30
Lys Ala Met His Asn Arg Leu Tyr Gly Met Asn Glu Glu Gly Trp Arg
35 40 45
Arg Ala Val Trp Glu Lys Asn Met Lys Met Ile Glu Leu His Asn Gln
50 55 60
Glu Tyr Arg Glu Gly Lys His Ser Phe Thr Met Ala Met Asn Ala Phe
65 70 75 80
Gly Asp Met Thr Ser Glu Glu Phe Arg Gln Val Met Asn Gly Phe Gln
85 90 95
Asn Arg Lys Pro Arg Lys Gly Lys Val Phe Gln Glu Pro Leu Phe Tyr
100 105 110
Glu Ala Pro Arg Ser Val Asp Trp Arg Glu Lys Gly Tyr Val Thr Pro
115 120 125
Val Lys Asn Gln Gly Gln Cys Gly Ser Cys Trp Ala Phe Ser Ala Thr
130 135 140
Gly Ala Leu Glu Gly Gln Met Phe Arg Lys Thr Gly Arg Leu Ile Ser
145 150 155 160
Leu Ser Glu Gln Asn Leu Val Asp Cys Ser Gly Pro Gln Gly Asn Glu
165 170 175
Gly Cys Asn Gly Gly Leu Met Asp Tyr Ala Phe Gln Tyr Val Gln Asp
180 185 190
Asn Gly Gly Leu Asp Ser Glu Glu Ser Tyr Pro Tyr Glu Ala Thr Glu
195 200 205
Glu Ser Cys Lys Tyr Asn Pro Lys Tyr Ser Val Ala Asn Asp Thr Gly
210 215 220
Phe Val Asp Ile Pro Lys Gln Glu Lys Ala Leu Met Lys Ala Val Ala
225 230 235 240
Thr Val Gly Pro Ile Ser Val Ala Ile Asp Ala Gly His Glu Ser Phe
245 250 255
Leu Phe Tyr Lys Glu Gly Ile Tyr Phe Glu Pro Asp Cys Ser Ser Glu
260 265 270
Asp Met Asp His Gly Val Leu Val Val Gly Tyr Gly Phe Glu Ser Thr
275 280 285
Glu Ser Asp Asn Asn Lys Tyr Trp Leu ValLys Asn Ser Trp Gly Glu
290 295 300
Glu Trp Gly Met Gly Gly Tyr Val Lys Met Ala Lys Asp Arg Arg Asn
305 310 315 320
His Cys Gly Ile Ala Ser Ala Ala Ser Tyr Pro Thr Val
325 330
<210>275
<211>1342
<212>DNA
<213>Homo sapiens
<220>
<223>Cathepsin V
<400>275
cggctgtaat ctcagaggct tgtttgctga gggtgcctgc gcacgtgcga cggctgctgg 60
ttttgaaaca tgaatctttc gctcgtcctg gctgcctttt gcttgggaat agcctccgct 120
gttccaaaat ttgaccaaaa tttggataca aagtggtacc agtggaaggc aacacacaga 180
agattatatg gcgcgaatga agaaggatgg aggagagcag tgtgggaaaa gaatatgaaa 240
atgattgaac tgcacaatgg ggaatacagc caagggaaac atggcttcac aatggccatg 300
aatgcttttc ctgacatgac caatgaagaa ttcaggcaga tgatgggttg ctttcgaaac 360
cagaaattca ggaaggggaa agtgttccgt gagcctctgt ttcttgatct tcccaaatct 420
gtggattgga gaaagaaagg ctacgtgacg ccagtgaaga atcagaaaca gtgtggttct 480
tgttgggctt ttagtgcgac tggtgctctt gaaggacaga tgttccggaa aactgggaaa 540
cttgtctcac tgagcgagca gaatctggtg gactgttcgc gtcctcaagg caatcagggc 600
tgcaatggtg gcttcatggc tagggccttc cagtatgtca aggagaacgg aggcctggac 660
tctgaggaat cctatccata tgtagcagtg gatgaaatct gtaagtacag acctgagaat 720
tctgttgcta atgacactgg cttcacagtg gtcgcacctg gaaaggagaa ggccctgatg 780
aaagcagtcg caactgtggg gcccatctcc gttgctatgg atgcaggcca ttcgtccttc 840
cagttctaca aatcaggcat ttattttgaa ccagactgca gcagcaaaaa cctggatcat 900
ggtgttctgg tggttggcta cggctttgaa ggagcaaatt cgaataacag caagtattgg 960
ctcgtcaaaa acagctgggg tccagaatgg ggctcgaatg gctatgtaaa aatagccaaa 1020
gacaagaaca accactgtgg aatcgccaca gcagccagct accccaatgt gtgagctgat 1080
ggatggtgag gaggaaggac ttaaggacag catgtctggg gaaattttat cttgaaactg 1140
accaaacgct tattgtgtaa gataaaccag ttgaatcatt gaggatccaa gttgagattt 1200
taattctgtg acatttttac aagggtaaaa tgttaccact actttaatta ttgttataca 1260
cagctttatg atatcaaaga ctcattgctt aattctaaga cttttgaatt ttcatttttt 1320
aaaaagatgt acaaaacagt tt 1342
<210>276
<211>334
<212>PRT
<213>Homo sapiens
<220>
<223>Cathepsin V
<400>276
Met Asn Leu Ser Leu Val Leu Ala Ala Phe Cys Leu Gly Ile Ala Ser
1 5 10 15
Ala Val Pro Lys Phe Asp Gln Asn Leu Asp Thr Lys Trp Tyr Gln Trp
20 25 30
Lys Ala Thr His Arg Arg Leu Tyr Gly Ala Asn Glu Glu Gly Trp Arg
35 40 45
Arg Ala Val Trp Glu Lys Asn Met Lys Met Ile Glu Leu His Asn Gly
50 55 60
Glu Tyr Ser Gln Gly Lys His Gly Phe Thr Met Ala Met Asn Ala Phe
65 70 75 80
Gly Asp Met Thr Asn Glu Glu Phe Arg Gln Met Met Gly Cys Phe Arg
85 90 95
Asn Gln Lys Phe Arg Lys Gly Lys Val Phe Arg Glu Pro Leu Phe Leu
100 105 110
Asp Leu Pro Lys Ser Val Asp Trp Arg Lys Lys Gly Tyr Val Thr Pro
115 120 125
Val Lys Asn Gln Lys Gln Cys Gly Ser Cys Trp Ala Phe Ser Ala Thr
130 135 140
Gly Ala Leu Glu Gly Gln Met Phe Arg Lys Thr Gly Lys Leu Val Ser
145 150 155 160
Leu Ser Glu Gln Asn Leu Val Asp Cys Ser Arg Pro Gln Gly Asn Gln
165 170 175
Gly Cys Asn Gly Gly Phe Met Ala Arg Ala Phe Gln Tyr Val Lys Glu
180 185 190
Asn Gly Gly Leu Asp Ser Glu Glu Ser Tyr Pro Tyr Val Ala Val Asp
195 200 205
Glu Ile Cys Lys Tyr Arg Pro Glu Asn Ser Val Ala Asn Asp Thr Gly
210 215 220
Phe Thr Val Val Ala Pro Gly Lys Glu Lys Ala Leu Met Lys Ala Val
225 230 235 240
Ala Thr Val Gly Pro Ile Ser Val Ala Met Asp Ala Gly His Ser Ser
245 250 255
Phe Gln Phe Tyr Lys Ser Gly Ile Tyr Phe Glu Pro Asp Cys Ser Ser
260 265 270
Lys Asn Leu Asp His Gly Val Leu Val Val Gly Tyr Gly Phe Glu Gly
275 280 285
Ala Asn Ser Asn Asn Ser Lys Tyr Trp Leu Val Lys Asn Ser Trp Gly
290 295 300
Pro Glu Trp Gly Ser Asn Gly Tyr Val Lys Ile Ala Lys Asp Lys Asn
305 310 315 320
Asn His Cys Gly Ile Ala Thr Ala Ala Ser Tyr Pro Asn Val
325 330
<210>277
<211>1661
<212>DNA
<213>Homo sapiens
<220>
<223>Cathepsin K
<400>277
attcggcacg agccgcaatc ccgatggaat aaatctagca cccctgatgg tgtgcccaca 60
ctttgctgcc gaaacgaagc cagacaacag atttccatca gcaggatgtg ggggctcaag 120
gttctgctgc tacctgtggt gagctttgct ctgtaccctg aggagatact ggacacccac 180
tgggagctat ggaagaagac ccacaggaag caatataaca acaaggtgga tgaaatctct 240
cggcgtttaa tttgggaaaa aaacctgaag tatatttcca tccataacct tgaggcttct 300
cttggtgtcc atacatatga actggctatg aaccacctgg gggacatgac cagtgaagag 360
gtggttcaga agatgactgg actcaaagta cccctgtctc attcccgcag taatgacacc 420
ctttatatcc cagaatggga aggtagagcc ccagactctg tcgactatcg aaagaaagga 480
tatgttactc ctgtcaaaaa tcagggtcag tgtggttcct gttgggcttt tagctctgtg 540
ggtgccctgg agggccaact caagaagaaa actggcaaac tcttaaatct gagtccccag 600
aacctagtgg attgtgtgtc tgagaatgat ggctgtggag ggggctacat gaccaatgcc 660
ttccaatatg tgcagaagaa ccggggtatt gactctgaag atgcctaccc atatgtggga 720
caggaagaga gttgtatgta caacccaaca ggcaaggcag ctaaatgcag agggtacaga 780
gagatccccg aggggaatga gaaagccctg aagagggcag tggcccgagt gggacctgtc 840
tctgtggcca ttgatgcaag cctgacctcc ttccagtttt acagcaaagg tgtgtattat 900
gatgaaagct gcaatagcga taatctgaac catgcggttt tggcagtggg atatggaatc 960
cagaagggaa acaagcactg gataattaaa aacagctggg gagaaaactg gggaaacaaa 1020
ggatatatcc tcatggctcg aaataagaac aacgcctgtg gcattgccaa cctggccagc 1080
ttccccaaga tgtgactcca gccagccaaa tccatcctgc tcttccattt cttccacgat 1140
ggtgcagtgt aacgatgcac tttggaaggg agttggtgtg ctatttttga agcagatgtg 1200
gtgatactga gattgtctgt tcagtttccc catttgtttg tgcttcaaat gatccttcct 1260
actttggttc tctcctccca tgaccttttt cactgtggcc atcaggactt tccctgacag 1320
ctgtgtactc ttaggctaag agatgtgact acagcctgcc cctgactgtg ttgtcccagg 1380
gctgatgctg tacaggtaca ggctggagat tttcacatag gttagattct cattcacggg 1440
actagttagc tttaagcacc ctagaggact agggtaatct gacttctcac ttcctaagtt 1500
cccttcaata tcctcaaggt agaaatgtct atgttttcta ctccaattca taaatctatt 1560
cataagtctt tggtacaagt ttacatgata aaaagaaatg tgatttgtct tcccttcttt 1620
gcacttttga aataaagtat ttatctcctg tctacagttt a 1661
<210>278
<211>329
<212>PRT
<213>Homo sapiens
<220>
<223>Cathepsin K
<400>278
Met Trp Gly Leu Lys Val Leu Leu Leu Pro Val Val Ser Phe Ala Leu
1 5 10 15
Tyr Pro Glu Glu Ile Leu Asp Thr His Trp Glu Leu Trp Lys Lys Thr
20 25 30
His Arg Lys Gln Tyr Asn Asn Lys Val Asp Glu Ile Ser Arg Arg Leu
35 40 45
Ile Trp Glu Lys Asn Leu Lys Tyr Ile Ser Ile His Asn Leu Glu Ala
50 55 60
Ser Leu Gly Val His Thr Tyr Glu Leu Ala Met Asn His Leu Gly Asp
65 70 75 80
Met Thr Ser Glu Glu Val ValGln Lys Met Thr Gly Leu Lys Val Pro
85 90 95
Leu Ser His Ser Arg Ser Asn Asp Thr Leu Tyr Ile Pro Glu Trp Glu
100 105 110
Gly Arg Ala Pro Asp Ser Val Asp Tyr Arg Lys Lys Gly Tyr Val Thr
115 120 125
Pro Val Lys Asn Gln Gly Gln Cys Gly Ser Cys Trp Ala Phe Ser Ser
130 135 140
Val Gly Ala Leu Glu Gly Gln Leu Lys Lys Lys Thr Gly Lys Leu Leu
145 150 155 160
Asn Leu Ser Pro Gln Asn Leu Val Asp Cys Val Ser Glu Asn Asp Gly
165 170 175
Cys Gly Gly Gly Tyr Met Thr Asn Ala Phe Gln Tyr Val Gln Lys Asn
180 185 190
Arg Gly Ile Asp Ser Glu Asp Ala Tyr Pro Tyr Val Gly Gln Glu Glu
195 200 205
Ser Cys Met Tyr Asn Pro Thr Gly Lys Ala Ala Lys Cys Arg Gly Tyr
210 215 220
Arg Glu Ile Pro Glu Gly Asn Glu Lys Ala Leu Lys Arg Ala Val Ala
225 230 235 240
Arg Val Gly Pro Val Ser Val Ala Ile Asp Ala Ser Leu Thr Ser Phe
245 250 255
Gln Phe Tyr Ser Lys Gly Val Tyr Tyr Asp Glu Ser Cys Asn Ser Asp
260 265 270
Asn Leu Asn His Ala Val Leu Ala Val Gly Tyr Gly Ile Gln Lys Gly
275 280 285
Asn Lys His Trp Ile Ile Lys Asn Ser Trp Gly Glu Asn Trp Gly Asn
290 295 300
Lys Gly Tyr Ile Leu Met Ala Arg Asn Lys Asn Asn Ala Cys Gly Ile
305 310 315 320
Ala Asn Leu Ala Ser Phe Pro Lys Met
325
<210>279
<211>1255
<212>DNA
<213>Homo sapiens
<220>
<223>Cathepsin S
<400>279
ctaaagatga aacggctggt ttgtgtgctc ttggtgtgct cctctgcagt ggcacagttg 60
cataaagatc ctaccctgga tcaccactgg catctctgga agaaaaccta tggcaaacaa 120
tacaaggaaa agaatgaaga agcagtacga cgtctcatct gggaaaagaa tctaaagttt 180
gtgatgcttc acaacctgga gcattcaatg ggaatgcact catacgatct gggcatgaac 240
cacctgggag acatgaccag tgaagaagtg atgtctttga cgagttccct gagagttccc 300
agccagtggc agagaaatat cacatataag tcaaacccta atcggatatt gcctgattct 360
gtggactgga gagagaaagg gtgtgttact gaagtgaaat atcaaggttc ttgtggtgct 420
tgctgggctt tcagtgctgt gggggccctg gaagcacagc tgaagctgaa aacaggaaag 480
ctggtgactc tcagtgccca gaacctggtg gattgctcaa ctgaaaaata tggaaacaaa 540
ggctgcaatg gtggcttcat gacaacggct ttccagtaca tcattgataa caagggcatc 600
gactcagacg cttcctatcc ctacaaagcc atggatcaga aatgtcaata tgactcaaaa 660
tatcgtgctg ccacatgttc aaagtacact gaacttcctt atgggagaga agatgtcctg 720
aaagaagctg tggccaataa aggcccagtg tctgttggtg tagatgcgcg tcatccttct 780
ttcttcctct acagaagtgg tgtctactat gaaccatcct gtactcagaa tgtgaatcat 840
ggtgtacttg tggttggcta tggtgatctt aatgggaaag aatactggct tgtgaaaaac 900
agctggggcc acaactttgg tgaagaagga tatattcgga tggcaagaaa taaaggaaat 960
cattgtggga ttgctagctt tccctcttac ccagaaatct agaggatctc tcctttttat 1020
aacaaatcaa gaaatatgaa gcactttctc ttaacttaat ttttcctgct gtatccagaa 1080
gaaataattg tgtcatgatt aatgtgtatt tactgtacta atagaaaata tagtttgagg 1140
ccgggcactg tctggctcac gcctgtaatc ccagtacttg ggaggccaag gaggcatatc 1200
aacttgaggc caggagttaa agagcagcct ggctaactgt gaaaccctct ctact 1255
<210>280
<211>331
<212>PRT
<213>Homo sapiens
<220>
<223>Cathepsin S
<400>280
Met Lys Arg Leu Val Cys Val Leu Leu Val Cys Ser Ser Ala Val Ala
1 5 10 15
Gln Leu His Lys Asp Pro Thr Leu Asp His His Trp His Leu Trp Lys
20 25 30
Lys Thr Tyr Gly Lys Gln Tyr Lys Glu Lys Asn Glu Glu Ala Val Arg
35 40 45
Arg Leu Ile Trp Glu Lys Asn Leu Lys Phe Val Met Leu His Asn Leu
50 55 60
Glu His Ser Met Gly Met His Ser Tyr Asp Leu Gly Met Asn His Leu
65 70 75 80
Gly Asp Met Thr Ser Glu Glu Val Met Ser Leu Met Ser Ser Leu Arg
85 90 95
Val Pro Ser Gln Trp Gln Arg Asn Ile Thr Tyr Lys Ser Asn Pro Asn
100 105 110
Arg Ile Leu Pro Asp Ser Val Asp Trp Arg Glu Lys Gly Cys Val Thr
115 120 125
Glu Val Lys Tyr Gln Gly Ser Cys Gly Ala Cys Trp Ala Phe Ser Ala
130 135 140
Val Gly Ala Leu Glu Ala Gln Leu Lys Leu Lys Thr Gly Lys Leu Val
145 150 155 160
Thr Leu Ser Ala Gln Asn Leu Val Asp Cys Ser Thr Glu Lys Tyr Gly
165 170 175
Asn Lys Gly Cys Asn Gly Gly Phe Met Thr Thr Ala Phe Gln Tyr Ile
180 185 190
Ile Asp Asn Lys Gly Ile Asp Ser Asp Ala Ser Tyr Pro Tyr Lys Ala
195 200 205
Met Asp Gln Lys Cys Gln Tyr Asp Ser Lys Tyr Arg Ala Ala Thr Cys
210 215 220
Ser Lys Tyr Thr Glu Leu Pro Tyr Gly Arg Glu Asp Val Leu Lys Glu
225 230 235 240
Ala Val Ala Asn Lys Gly Pro Val Ser Val Gly Val Asp Ala Arg His
245 250 255
Pro Ser Phe Phe Leu Tyr Arg Ser Gly Val Tyr Tyr Glu Pro Ser Cys
260 265 270
Thr Gln Asn Val Asn His Gly Val Leu Val Val Gly Tyr Gly Asp Leu
275 280 285
Asn Gly Lys Glu Tyr Trp Leu ValLys Asn Ser Trp Gly His Asn Phe
290 295 300
Gly Glu Glu Gly Tyr Ile Arg Met Ala Arg Asn Lys Gly Asn His Cys
305 310 315 320
Gly Ile Ala Ser Phe Pro Ser Tyr Pro Glu Ile
325 330
<210>281
<211>1517
<212>DNA
<213>Homo sapiens
<220>
<223>Cathepsin F
<400>281
gaattccggg tctgttgggt ccgggccgcc ggcttcgccc tcgccatggc gccctggctg 60
cagctcctgt cgctgctggg gctgctcccg ggcgcagtgg ccgcccccgc ccagccccga 120
gccgccagct ttcaggcctg ggggccgccg tccccggagc tgctggcgcc cacccgcttc 180
gcgctggaga tgttcaaccg cggccgggct gcggggacgc gggccgtgctgggccttgtg 240
cgcggccgcg tccgccgggc gggtcagggg tcgctgtact ccctggaggc caccctggag 300
gagccaccct gcaacgaccc catggtgtgc cggctccccg tgtccaagaa aaccctgctc 360
tgcagcttcc aagtcctgga tgagctcgga agacacgtgc tgctgcggaa ggactgtggc 420
ccagtggaca ccaaggttcc aggtgctggg gagcccaagt cagccttcac tcagggctca 480
gccatgattt cttctctgtc ccaaaaccat ccagacaaca gaaacgagac tttcagctca 540
gtcatttccc tgttgaatga ggatcccctg tcccaggact tgcctgtgaa gatggcttca 600
atcttcaaga actttgtcat tacctataac cggacatatg agtcaaagga agaagcccgg 660
tggcgcctgt ccgtctttgt caataacatg gtgcgagcac agaagatcca ggccctggac 720
cgtgggacag ctcagtatgg agtcaccaag ttcagtgatc tcacagagga ggagttccgc 780
actatctacc tgaatactct cctgaggaaa gagcctggca acaagatgaa gcaagccaag 840
tctgtgggtg acctcgcccc acctgaatgg gactggagga gtaagggggc tgtcacaaaa 900
gtcaaagacc agggcatgtg tggctcctgc tgggccttct cagtcacagg caatgtggag 960
ggccagtggt ttctcaacca ggggaccctg ctctccctct ctgaacagga gctcttggac 1020
tgtgacaaga tggacaaggc ctgcatgggc ggcttgccct ccaatgccta ctcggccata 1080
aagaatttgg gagggctgga gacagaggat gactacagct accagggtca catgcagtcc 1140
tgcaacttct cagcagagaa ggccaaggtc tacatcaatg actccgtgga gctgagccag 1200
aacgagcaga agctggcagc ctggctggcc aagagaggcc caatctccgt ggccatcaat 1260
gcctttggca tgcagtttta ccgccacggg atctcccgcc ctctccggcc cctctgcagc 1320
ccttggctca ttgaccatgc ggtgttgctt gtgggctacg gcaaccgctc tgacgttccc 1380
ttttgggcca tcaagaacag ctggggcact gactggggtg agaagggtta ctactacttg 1440
catcgtgggt ccggggcctg tggcgtgaac accatggcca gctcggcggt ggtggactga 1500
agaggggccc cgaattc 1517
<210>282
<211>484
<212>PRT
<213>Homo sapiens
<220>
<223>Cathepsin F
<400>282
Met Ala Pro Trp Leu Gln Leu Leu Ser Leu Leu Gly Leu Leu Pro Gly
1 5 10 15
Ala Val Ala Ala Pro Ala Gln Pro Arg Ala Ala Ser Phe Gln Ala Trp
20 25 30
Gly Pro Pro Ser Pro Glu Leu Leu Ala Pro Thr Arg Phe Ala Leu Glu
35 40 45
Met Phe Asn Arg Gly Arg Ala Ala Gly Thr Arg Ala Val Leu Gly Leu
50 55 60
Val Arg Gly Arg Val Arg Arg Ala Gly Gln Gly Ser Leu Tyr Ser Leu
65 70 75 80
Glu Ala Thr Leu Glu Glu Pro Pro Cys Asn Asp Pro Met Val Cys Arg
85 90 95
Leu Pro Val Ser Lys Lys Thr Leu Leu Cys Ser Phe Gln Val Leu Asp
100 105 110
Glu Leu Gly Arg His Val Leu Leu Arg Lys Asp Cys Gly Pro Val Asp
115 120 125
Thr Lys Val Pro Gly Ala Gly Glu Pro Lys Ser Ala Phe Thr Gln Gly
130 135 140
Ser Ala Met Ile Ser Ser Leu Ser Gln Asn His Pro Asp Asn Arg Asn
145 150 155 160
Glu Thr Phe Ser Ser Val Ile Ser Leu Leu Asn Glu Asp Pro Leu Ser
165 170 175
Gln Asp Leu Pro Val Lys Met Ala Ser Ile Phe Lys Asn Phe Val Ile
180 185 190
Thr Tyr Asn Arg Thr Tyr Glu Ser Lys Glu Glu Ala Arg Trp Arg Leu
195 200 205
Ser Val Phe Val Asn Asn Met Val Arg Ala Gln Lys Ile Gln Ala Leu
210 215 220
Asp Arg Gly Thr Ala Gln Tyr Gly Val Thr Lys Phe Ser Asp Leu Thr
225 230 235 240
Glu Glu Glu Phe Arg Thr Ile Tyr Leu Asn Thr Leu Leu Arg Lys Glu
245 250 255
Pro Gly Asn Lys Met Lys Gln Ala Lys Ser Val Gly Asp Leu Ala Pro
260 265 270
Pro Glu Trp Asp Trp Arg Ser Lys Gly Ala Val Thr Lys Val Lys Asp
275 280 285
Gln Gly Met Cys Gly Ser Cys Trp Ala Phe Ser Val Thr Gly Asn Val
290 295 300
Glu Gly Gln Trp Phe Leu Asn Gln Gly Thr Leu Leu Ser Leu Ser Glu
305 310 315 320
Gln Glu Leu Leu Asp Cys Asp Lys Met Asp Lys Ala Cys Met Gly Gly
325 330 335
Leu Pro Ser Asn Ala Tyr Ser Ala Ile Lys Asn Leu Gly Gly Leu Glu
340 345 350
Thr Glu Asp Asp Tyr Ser Tyr Gln Gly His Met Gln Ser Cys Asn Phe
355 360 365
Ser Ala Glu Lys Ala Lys Val Tyr Ile Asn Asp Ser Val Glu Leu Ser
370 375 380
Gln Asn Glu Gln Lys Leu Ala Ala Trp Leu Ala Lys Arg Gly Pro Ile
385 390 395 400
Ser Val Ala Ile Asn Ala Phe Gly Met Gln Phe Tyr Arg His Gly Ile
405 410 415
Ser Arg Pro Leu Arg Pro Leu Cys Ser Pro Trp Leu Ile Asp His Ala
420 425 430
Val Leu Leu Val Gly Tyr Gly Asn Arg Ser Asp Val Pro Phe Trp Ala
435 440 445
Ile Lys Asn Ser Trp Gly Thr Asp Trp Gly Glu Lys Gly Tyr Tyr Tyr
450 455 460
Leu His Arg Gly Ser Gly Ala Cys Gly ValAsn Thr Met Ala Ser Ser
465 470 475 480
Ala Val Val Asp
<210>283
<211>2002
<212>DNA
<213>Homo sapiens
<220>
<223>Cathepsin B
<400>283
aattccgcgg caaccgctcc ggcaacgcca accgctccgc tgcgcgcagg ctgggctgca 60
ggctctcggc tgcagcgctg ggctggtgtg cagtggtgcg accacggctc acggcagcct 120
cagccaccca gatgtaagcg atctggttcc cacctcagcc ttccgagtag tggatctagg 180
atctggcttc caacatgtgg cagctctggg cctccctctg ctgcctgctg gtgttggcca 240
atgcccggag caggccctct ttccatcccg tgtcggatga gctggtcaac tatgtcaaca 300
aacggaatac cacgtggcag gccgggcaca acttctacaa cgtggacatg agctacttga 360
agaggctatg tggtaccttc ctgggtgggc ccaagccacc ccagagagtt atgtttaccg 420
aggacctgaa gctgcctgca agcttcgatg cacgggaaca atggccacag tgtcccacca 480
tcaaagagat cagagaccag ggctcctgtg gctcctgctg ggccttcggg gctgtggaag 540
ccatctctga ccgcatctgc atccacacca atgcgcacgt cagcgtggag gtgtcggcgg 600
aggacctgct cacctgctgt ggcagcatgt gtggggacgg ctgtaatggt ggctatcctg 660
ctgaagcttg gaacttctgg acaagaaaag gcctggtttc tggtggcctc tatgaatccc 720
atgtagggtg cagaccgtac tccatccctc cctgtgagca ccacgtcaac ggctcccggc 780
ccccatgcac gggggaggga gataccccca agtgtagcaa gatctgtgag cctggctaca 840
gcccgaccta caaacaggac aagcactacg gatacaattc ctacagcgtc tccaatagcg 900
agaaggacat catggccgag atctacaaaa acggccccgt ggagggagct ttctctgtgt 960
attcggactt cctgctctac aagtcaggag tgtaccaaca cgtcaccgga gagatgatgg 1020
gtggccatgc catccgcatc ctgggctggg gagtggagaa tggcacaccc tactggctgg 1080
ttgccaactc ctggaacact gactggggtg acaatggctt ctttaaaata ctcagaggac 1140
aggatcactg cggaatcgaa tcagaagtgg tggctggaat tccacgcacc gatcagtact 1200
gggaaaagat ctaatctgcc gtgggcctgt cgtgccagtc ctgggggcga gatcggggta 1260
gaaagtcatt ttattcttta agttcacgta agatacaagt ttcaggcagg gtctgaagga 1320
ctggattggc caaagtcctc caaggagacc aagtcctggc tacatcccag cctgtggtta 1380
cagtgcagac aggccatgtg agccaccgct gccagcacag agcgtccttc cccctgtaga 1440
ctagtgccgt gggagtacct gctgcccagc tgctgtggcc ccctccgtga tccatccatc 1500
tccagggagc aagacagaga cgcaggatgg aaagcggagt tcctaacagg atgaaagttc 1560
ccccatcagt tcccccagta cctccaagca agtagctttc cacatttgtc acagaaatca 1620
gaggagagat ggtgttggga gccctttgga gaacgccagt ctccaggtcc ccctgcatct 1680
atcgagtttg caatgtcaca acctctctga tcttgtgctc agcatgattc tttaatagaa 1740
gttttatttt tcgtgcactc tgctaatcat gtgggtgagc cagtggaaca gcgggagcct 1800
gtgctggttt gcagattgcc tcctaatgac gcggctcaaa aggaaaccaa gtggtcagga 1860
gttgtttctg acccactgat ctctactacc acaaggaaaa tagtttagga gaaaccagct 1920
tttactgttt ttgaaaaatt acagcttcac cctgtcaagt taacaaggaa tgcctgtgcc 1980
aataaaaggt ttctccaact tg 2002
<210>284
<211>339
<212>PRT
<213>Homo sapiens
<220>
<223>Cathepsin B
<400>284
get Trp Gln Leu Trp Ala Ser Leu Cys Cys Leu Leu Val Leu Ala Asn
1 5 10 15
Ala Arg Ser Arg Pro Ser Phe His Pro Leu Ser Asp Glu Leu Val Asn
20 25 30
Tyr Val Asn Lys Arg Asn Thr Thr Trp Gln Ala Gly His Asn Phe Tyr
35 40 45
Asn Val Asp Met Ser Tyr Leu Lys Arg Leu Cys Gly Thr Phe Leu Gly
50 55 60
Gly Pro Lys Pro Pro Gln Arg Val Met Phe Thr Glu Asp Leu Lys Leu
65 70 75 80
Pro Ala Ser Phe Asp Ala Arg Glu Gln Trp Pro Gln Cys Pro Thr Ile
85 90 95
Lys Glu Ile Arg Asp Gln Gly Ser Cys Gly Ser Cys Trp Ala Phe Gly
100 105 110
Ala Val Glu Ala Ile Ser Asp Arg Ile Cys Ile His Thr Asn Ala His
115 120 125
Val Ser Val Glu Val Ser Ala Glu Asp Leu Leu Thr Cys Cys Gly Ser
130 135 140
Met Cys Gly Asp Gly Cys Asn Gly Gly Tyr Pro Ala Glu Ala Trp Asn
145 150 155 160
Phe Trp Thr Arg Lys Gly Leu Val Ser Gly Gly Leu Tyr Glu Ser His
165 170 175
Val Gly Cys Arg Pro Tyr Ser Ile Pro Pro Cys Glu His His Val Asn
180 185 190
Gly Ser Arg Pro Pro Cys Thr Gly Glu Gly Asp Thr Pro Lys Cys Ser
195 200 205
Lys Ile Cys Glu Pro Gly Tyr Ser Pro Thr Tyr Lys Gln Asp Lys His
210 215 220
Tyr Gly Tyr Asn Ser Tyr Ser Val Ser Asn Ser Glu Lys Asp Ile Met
225 230 235 240
Ala Glu Ile Tyr Lys Asn Gly Pro Val Glu Gly Ala Phe Ser Val Tyr
245 250 255
Ser Asp Phe Leu Leu Tyr Lys Ser Gly Val Tyr Gln His Val Thr Gly
260 265 270
Glu Met Met Gly Gly His Ala Ile Arg Ile Leu Gly Trp Gly Val Glu
275 280 285
Asn Gly Thr Pro Tyr Trp Leu Val Ala Asn Ser Trp Asn Thr Asp Trp
290 295 300
Gly Asp Asn Gly Phe Phe Lys Ile Leu Arg Gly Gln Asp His Cys Gly
305 310 315 320
Ile Glu Ser Glu Val Val Ala Gly Ile Pro Arg Thr Asp Gln Tyr Trp
325 330 335
Glu Lys Ile
<210>285
<211>1292
<212>DNA
<213>Carica papaya
<220>
<223>Papain
<400>285
atccattccc acttaagaag taaaaagata tagctagtgt cacaatggct atgatacctt 60
caatttcaaa gttgcttttt gttgcaatat gtctttttgt ttatatgggt ttgtcatttg 120
gtgatttttc tattgtgggt tattctcaaa atgacttgac atccactgaa agacttattc 180
agctatttga atcgtggatg ttgaagcaca ataagattta taagaatatt gatgaaaaaa 240
tctacagatt tgaaattttt aaagataatc ttaaatatat tgatgagaca aataagaaaa 300
ataacagtta ttggcttgga ttaaatgtgt ttgctgatat gagcaatgat gaattcaaag 360
aaaagtatac tggttctatt gctggaaatt atacaacaac cgaactatca tacgaagaag 420
tgcttaatga tggtgatgta aatatcccgg agtatgtcga ttggagacaa aaaggagctg 480
tcactcctgt aaaaaatcag ggttcttgtg gtagttgttg ggcattctca gctgttgtaa 540
ctatagaggg aataattaag attagaactg ggaacttaaa tgaatactca gagcaagaac 600
tgcttgactg cgacagacgt agctacgggt gcaatggagg ttacccttgg agtgcacttc 660
aattagtggc tcaatatggt attcactaca gaaatactta cccatatgag ggagtgcaac 720
gttattgtcg ctcaagggag aaaggtcctt atgcagccaa aaccgatggg gttcgacaag 780
tgcaaccata taatgaaggg gctctcttat attcaattgc aaaccaacct gtgagcgttg 840
tccttgaagc tgctggaaaa gatttccaat tatatagagg gggaatattt gtggggccat 900
gcggaaacaa agtagatcat gcagtcgcag cagttgggta tggaccaaat tacatactca 960
taaagaattc atggggtaca ggatggggtg aaaatggata tataagaatc aaaagaggca 1020
ctggaaactc ctatggagta tgtggacttt atacaagctc attctatcct gttaaaaact 1080
gatgagatca cggctttcat aaaatccctt atatatatat atatatatag aacttgtata 1140
ctcatcgtgt gttgaaataa taaatgagag gattaataat ttgtgtaatc ctatatatca 1200
cagtttgttg tgacaaactt ttgcatcgct tgttatatta tttgtaataa tgtttgtttt 1260
gattgaataa acttttacat atacttttat gc 1292
<210>286
<211>345
<212>PRT
<213>Carica papaya
<220>
<223>Papain
<400>286
Met Ala Met Ile Pro Ser Ile Ser Lys Leu Leu Phe Val Ala Ile Cys
1 5 10 15
Leu Phe Val Tyr Met Gly Leu Ser Phe Gly Asp Phe Ser Ile Val Gly
20 25 30
Tyr Ser Gln Asn Asp Leu Thr Ser Thr Glu Arg Leu Ile Gln Leu Phe
35 40 45
Glu Ser Trp Met Leu Lys His Asn Lys Ile Tyr Lys Asn Ile Asp Glu
50 55 60
Lys Ile Tyr Arg Phe Glu Ile Phe Lys Asp Asn Leu Lys Tyr Ile Asp
65 70 75 80
Glu Thr Asn Lys Lys Asn Asn Ser Tyr Trp Leu Gly Leu Asn Val Phe
85 90 95
Ala Asp Met Ser Asn Asp Glu Phe Lys Glu Lys Tyr Thr Gly Ser Ile
100 105 110
Ala Gly Asn Tyr Thr Thr Thr Glu Leu Ser Tyr G1u Glu Val Leu Asn
115 120 125
Asp Gly Asp Val Asn Ile Pro Glu Tyr Val Asp Trp Arg Gln Lys Gly
130 135 140
Ala Val Thr Pro Val Lys Asn Gln Gly Ser Cys Gly Ser Cys Trp Ala
145 150 155 160
Phe Ser Ala Val Val Thr Ile Glu Gly Ile Ile Lys Ile Arg Thr Gly
165 170 175
Asn Leu Asn Glu Tyr Ser Glu Gln Glu Leu Leu Asp Cys Asp Arg Arg
180 185 190
Ser Tyr Gly Cys Asn Gly Gly Tyr Pro Trp Ser Ala Leu Gln Leu Val
195 200 205
Ala Gln Tyr Gly Ile His Tyr Arg Asn Thr Tyr Pro Tyr Glu Gly Val
210 215 220
Gln Arg Tyr Cys Arg Ser Arg Glu Lys Gly Pro Tyr Ala Ala Lys Thr
225 230 235 240
Asp Gly Val Arg Gln Val Gln Pro Tyr Asn Glu Gly Ala Leu Leu Tyr
245 250 255
Ser Ile Ala Asn Gln Pro Val Ser Val Val Leu Glu Ala Ala Gly Lys
260 265 270
Asp Phe Gln Leu Tyr Arg Gly Gly Ile Phe Val Gly Pro Cys Gly Asn
275 280 285
Lys Val Asp His Ala Val Ala Ala Val Gly Tyr Gly Pro Asn Tyr Ile
290 295 300
Leu Ile Lys Asn Ser Trp Gly Thr Gly Trp Gly Glu Asn Gly Tyr Ile
305 310 315 320
Arg Ile Lys Arg Gly Thr Gly Asn Ser Tyr Gly Val Cys Gly Leu Tyr
325 330 335
Thr Ser Ser Phe Tyr Pro Val Lys Asn
340 345
<210>287
<211>1845
<212>DNA
<213>Trypanosoma cruzi
<220>
<223>Cruzain (Cruzapain)
<400>287
atgtctggct gggcgcgtgc gctgttgctc gcggccgtcc tggtcgtcat ggcgtgcctc 60
gtccccgcgg cgacggcgag cctgcatgcg gaggagacgc tgacgtcgca attcgcagaa 120
ttcaagcaga agcatggcag ggtgtacgag agcgccgcgg aggaggcgtt ccgcctgagc 180
gtgttcaggg agaacctgtt tcttgcgagg ctgcacgccg cggcaaaccc acacgcgacc 240
ttcggcgtca cgcccttctc ggacctcacg cgcgaggaat tccggtcccg ctaccacaac 300
ggcgcggcgc actttgcggc ggcgcaggag cgcgcgagag tgccggtgaa ggtggaggta 360
gttggcgcgc ccgcggcagt ggattggcgt gcgagaggcg ccgtgacagc cgtcaaggac 420
cagggccaat gcggttcgtg ctgggccttc tccgccattg gcaacgttga gtgccagtgg 480
tttcttgccg gccacccgct gacgaacctg tcggagcaga tgctcgtgtc gtgcgacaaa 540
acggactctg gctgcagtgg tggcctgatg aacaacgcct ttgagtggat tgtgcaggag 600
aataacggcg ccgtgtacac ggaggacagc tacccttatg cgtcgggcga ggggatatcg 660
ccgccgtgca cgacgtcagg ccacacggtg ggtgccacga ttaccggtca cgttgaatta 720
cctcaggacg aggcccaaat agccgcatgg cttgcagtca atggcccggt tgccgttgcc 780
gtcgacgcca gcagctggat gacctacacg ggcggcgtta tgacgagctg cgtctccgag 840
cagctggatc acggcgttct tctcgtcggc tacaatgaca gcgccgcagt gccgtactgg 900
atcatcaaga actcgtggac cacgcagtgg ggcgaggaag gctacatccg cattgcaaag 960
ggctcgaacc agtgccttgt caaggaggag gcgagctccg cggtggtcgg tggtcccgga 1020
cccactcccg agccaaccac cacgacaacc acaagtgccc caggaccgtc cccatcgtac 1080
tttgtgcaga tgtcctgcac tgacgctgcg tgcattgtcg ggtgcgagaa cgtgacgtta 1140
ccgaccggtc agtgtctcct gaccaccagc ggcgtctctg ccattgtcac gtgcggtgct 1200
gagactctca cagaagaagt cttccttacg agtacgcact gcagcggccc atcggtgagg 1260
tcctctgttc ctctcaacaa atgcaaccgg cttttaagag gctccgttga gttcttctgc 1320
ggctccagct ccagtggccg actggccgac gtggacaggc agcgtcgcca tcagccatac 1380
cacagccgtc atcgccgcct ctgagggcac tctttgtttc tgatgctgcc ttttctctca 1440
ttttactttt ttattattat tgagcgagcg tcggtgctgc ggcacactgg gcaaataagt 1500
cccagtggag tggattgttt attgatttgc tccatgtatg tgttttcttt tttgtgcaca 1560
actgcaaata tgtgtccgcg atcgacgtct cagccacgcg tgtgtttttt ctattatatg 1620
cttctgttga gcgcccgagc attccccact gtggctgaac ccaccgccgt ccccgactgt 1680
cacccgacca tgcggggcgg ggtggcgtgg caacgctcct ttgccgcggc gtgcacatac 1740
acgcgcgcat gcttcttctc tccacgtcaa cgccgacacc accacagtct gaaggtgacg 1800
ctgttgaagg cacacaagga aggaaacaca cccacacaag cagta 1845
<210>288
<211>467
<212>PRT
<213>Trypanosoma cruzi
<220>
<223>Cruzain (Cruzapain)
<400>288
Met Ser Gly Trp Ala Arg Ala Leu Leu Leu Ala Ala Val Leu Val Val
1 5 10 15
Met Ala Cys Leu Val Pro Ala Ala Thr Ala Ser Leu His Ala Glu Glu
20 25 30
Thr Leu Thr Ser Gln Phe Ala Glu Phe Lys Gln Lys His Gly Arg Val
35 40 45
Tyr Glu Ser Ala Ala Glu Glu Ala Phe Arg Leu Ser Val Phe Arg Glu
50 55 60
Asn Leu Phe Leu Ala Arg Leu His Ala Ala Ala Asn Pro His Ala Thr
65 70 75 80
Phe Gly Val Thr Pro Phe Ser Asp Leu Thr Arg Glu Glu Phe Arg Ser
85 90 95
Arg Tyr His Asn Gly Ala Ala His Phe Ala Ala Ala Gln Glu Arg Ala
100 105 110
Arg Val Pro Val Lys Val Glu Val Val Gly Ala Pro Ala Ala Val Asp
115 120 125
Trp Arg Ala Arg Gly Ala Val Thr Ala Val Lys Asp Gln Gly Gln Cys
130 135 140
Gly Ser Cys Trp Ala Phe Ser Ala Ile Gly Asn Val Glu Cys Gln Trp
145 150 155 160
Phe Leu Ala Gly His Pro Leu Thr Asn Leu Ser Glu Gln Met Leu Val
165 170 175
Ser Cys Asp Lys Thr Asp Ser Gly Cys Ser Gly Gly Leu Met Asn Asn
180 185 190
Ala Phe Glu Trp Ile Val Gln Glu Asn Asn Gly Ala Val Tyr Thr Glu
195 200 205
Asp Ser Tyr Pro Tyr Ala Ser Gly Glu Gly Ile Ser Pro Pro Cys Thr
210 215 220
Thr Ser Gly His Thr Val Gly Ala Thr Ile Thr Gly His Val Glu Leu
225 230 235 240
Pro Gln Asp Glu Ala Gln Ile Ala Ala Trp Leu Ala Val Asn Gly Pro
245 250 255
Val Ala Val Ala Val Asp Ala Ser Ser Trp Met Thr Tyr Thr Gly Gly
260 265 270
Val Met Thr Ser Cys Val Ser Glu Gln Leu Asp His Gly Val Leu Leu
275 280 285
Val Gly Tyr Asn Asp Ser Ala Ala Val Pro Tyr Trp Ile Ile Lys Asn
290 295 300
Ser Trp Thr Thr Gln Trp Gly Glu Glu Gly Tyr Ile Arg Ile Ala Lys
305 310 315 320
Gly Ser Asn Gln Cys Leu Val Lys Glu Glu Ala Ser Ser Ala Val Val
325 330 335
Gly Gly Pro Gly Pro Thr Pro Glu Pro Thr Thr Thr Thr Thr Thr Ser
340 345 350
Ala Pro Gly Pro Ser Pro Ser Tyr Phe Val Gln Met Ser Cys Thr Asp
355 360 365
Ala Ala Cys Ile Val Gly Cys Glu Asn ValThr Leu Pro Thr Gly Gln
370 375 380
Cys Leu Leu Thr Thr Ser Gly Val Ser Ala Ile ValThr Cys Gly Ala
385 390 395 400
Glu Thr Leu Thr Glu Glu Val Phe Leu Thr Ser Thr His Cys Ser Gly
405 410 415
Pro Ser Val Arg Ser Ser Val Pro Leu Asn Lys Cys Asn Arg Leu Leu
420 425 430
Arg Gly Ser ValGlu Phe Phe Cys Gly Ser Ser Ser Ser Gly Arg Leu
435 440 445
Ala Asp Val Asp Arg Gln Arg Arg His Gln Pro Tyr His Ser Arg His
450 455 460
Arg Arg Leu
465
<210>289
<211>379
<212>PRT
<213>Bacillus licheniformis
<220>
<223>Subtilisin Carlsberg precursor S08.001
<400>289
Met Met Arg Lys Lys Ser Phe Trp Leu Gly Met Leu Thr Ala Phe Met
1 5 10 15
Leu Val Phe Thr Met Ala Phe Ser Asp Ser Ala Ser Ala Ala Gln Pro
20 25 30
Ala Lys Asn Val Glu Lys Asp Tyr Ile ValGly Phe Lys Ser Gly Val
35 40 45
Lys Thr Ala Ser ValLys Lys Asp Ile Ile Lys Glu Ser Gly Gly Lys
50 55 60
Val Asp Lys Gln Phe Arg Ile Ile Asn Ala Ala Lys Ala Lys Leu Asp
65 70 75 80
Lys Glu Ala Leu Lys Glu Val Lys Asn Asp Pro Asp Val Ala Tyr Val
85 90 95
Glu Glu Asp His Val Ala His Ala Leu Ala Gln Thr Val Pro Tyr Gly
100 105 110
Ile Pro Leu Ile Lys Ala Asp Lys Val Gln Ala Gln Gly Phe Lys Gly
115 120 125
Ala Asn Val Lys Val Ala Val Leu Asp Thr Gly Ile Gln Ala Ser His
130 135 140
Pro Asp Leu Asn Val ValGly Gly Ala Ser Phe Val Ala Gly Glu Ala
145 150 155 160
Tyr Asn Thr Asp Gly Asn Gly His Gly Thr His Val Ala Gly Thr Val
165 170 175
Ala Ala Leu Asp Asn Thr Thr Gly Val Leu Gly Val Ala Pro Ser Val
180 185 190
Ser Leu Tyr Ala Val Lys Val Leu Asn Ser Ser Gly Ser Gly Thr Tyr
195 200 205
Ser Gly Ile Val Ser Gly Ile Glu Trp Ala Thr Thr Asn Gly Met Asp
210 215 220
Val Ile Asn Met Ser Leu Gly Gly Pro Ser Gly Ser Thr Ala Met Lys
225 230 235 240
Gln Ala Val Asp Asn Ala Tyr Ala Arg Gly Val Val Val Val Ala Ala
245 250 255
Ala Gly Asn Ser Gly Ser Ser Gly Asn Thr Asn Thr Ile Gly Tyr Pro
260 265 270
Ala Lys Tyr Asp Ser Val Ile Ala Val Gly Ala Val Asp Ser Asn Ser
275 280 285
Asn Arg Ala Ser Phe Ser Ser Val Gly Ala Glu Leu Glu Val Met Ala
290 295 300
Pro Gly Ala Gly Val Tyr Ser Thr Tyr Pro Thr Ser Thr Tyr Ala Thr
305 310 315 320
Leu Asn Gly Thr Ser Met Ala Ser Pro His Val Ala Gly Ala Ala Ala
325 330 335
Leu Ile Leu Ser Lys His Pro Asn Leu Ser Ala Ser Gln Val Arg Asn
340 345 350
Arg Leu Ser Ser Thr Ala Thr Tyr Leu Gly Ser Ser Phe Tyr Tyr Gly
355 360 365
Lys Gly Leu Ile Asn Val Glu Ala Ala Ala Gln
370 375
<210>290
<211>1588
<212>DNA
<213>Bacillus licheniformis
<220>
<223>Subtilisin Carlsberg precursor S08.001
<400>290
gatcgacaag accgcaacct ccttcgacaa aaaatgatct cttaaaataa atgaatagta 60
ttttcataaa atgaatcaga tggagcaatc tcctgtcatt cgcggccctc gggacctctt 120
tccctgccag gctgaagcgg tctattcata ctttcgaact gaacattttt ctaaaacagt 180
tattaataac caaaaaattt taaattggtc ctccaaaaaa ataggcctac catataattc 240
attttttttc tataataaat taacagaata attggaatag attatattat ccttctattt 300
aaattattct gaataaagag gaggagagtg agtaatgatg aggaaaaaga gtttttggct 360
tgggatgctg acggccttca tgctcgtgtt cacgatggca ttcagcgatt ccgcttctgc 420
tgctcaaccg gcgaaaaatg ttgaaaagga ttatattgtc ggatttaagt caggagtgaa 480
aaccgcatct gtcaaaaagg acatcatcaa agagagcggc ggaaaagtgg acaagcagtt 540
tagaatcatc aacgcggcaa aagcgaagct agacaaagaa gcgcttaagg aagtcaaaaa 600
tgatccggat gtcgcttatg tggaagagga tcatgtggcc catgccttgg cgcaaaccgt 660
tccttacggc attcctctca ttaaagcgga caaagtgcag gctcaaggct ttaagggagc 720
gaatgtaaaa gtagccgtcc tggatacagg aatccaagct tctcatccgg acttgaacgt 780
agtcggcgga gcaagctttg tggctggcga agcttataac accgacggca acggacacgg 840
cacacatgtt gccggtacag tagctgcgct tgacaataca acgggtgtat taggcgttgc 900
gccaagcgta tccttgtacg cggttaaagt actgaattca agcggaagcg gaacttacag 960
cggcattgta agcggaatcg agtgggcgac gacaaacggc atggatgtta tcaacatgag 1020
tcttggagga ccatcaggct caacagcgat gaaacaggcg gttgacaatg catatgcaag 1080
aggggttgtc gttgtggcgg ctgctgggaa cagcggatct tcaggaaaca cgaatacaat 1140
cggctatcct gcgaaatacg actctgtcat cgcagttggc gcggtagact ctaacagcaa 1200
cagagcttca ttttccagcg tcggagcaga gcttgaagtc atggctcctg gcgcaggcgt 1260
gtacagcact tacccaacca gcacttatgc aacattgaac ggaacgtcaa tggcttctcc 1320
tcatgtagcg ggagcagcag ctttgatctt gtcaaaacat ccgaaccttt cagcttcaca 1380
agtccgcaac cgtctctcca gtacggcgac ttatttggga agctccttct actatggaaa 1440
aggtctgatc aatgtcgaag ctgccgctca ataacatatt ctaacaaatg gcatatagaa 1500
aaagctagtg tttttagcac tagctttttc ttcattcagt tgaagactgt tcaatatttt 1560
gaatccgttc cattatggtc ggatggcc 1588
<210>291
<211>275
<212>PRT
<213>Bacillus pumilus
<220>
<223>Subtilisin(Alkaline mesentericopeptidase)S08.002
<400>291
Ala Gln Ser Val Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu
1 5 10 15
His Ser Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp
20 25 30
Ser Gly Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala
35 40 45
Ser Phe Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His
50 55 60
Gly Thr His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly
65 70 75 80
Val Leu Gly Val Ala Pro Ser Ser Ala Leu Tyr Ala Val Lys Val Leu
85 90 95
Asp Ser Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu
100 105 110
Trp Ala Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly
115 120 125
Pro Thr Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser
130 135 140
Ser Gly Ile Val Val Ala Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly
145 150 155 160
Ser Thr Ser Thr Val Gly Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala
165 170 175
Val Gly Ala Val Asn Ser Ala Asn Gln Arg Ala Ser Phe Ser Ser Ala
180 185 190
Gly Ser Glu Leu Asp Val Met Ala Pro Gly Val Ser Ile Gln Ser Thr
195 200 205
Leu Pro Gly Gly Thr Tyr Gly Ala Tyr Asn Gly Thr Ser Met Ala Thr
210 215 220
Pro His Val Ala Gly Ala Ala Ala Leu Ile Leu Ser Lys His Pro Thr
225 230 235 240
Trp Thr Asn Ala Gln Val Arg Asp Arg Leu Glu Ser Thr Ala Thr Tyr
245 250 255
Leu Gly Ser Ser Phe Tyr Tyr Gly Lys Gly Leu Ile Asn Val Gln Ala
260 265 270
Ala Ala Gln
275
<210>292
<211>269
<212>PRT
<213>Bacillus lentus
<220>
<223>Subtilisin Savinase (Alkaline protease)S08.003
<400>292
Ala Gln Ser Val Pro Trp Gly Ile Ser Arg Val Gln Ala Pro Ala Ala
1 5 10 15
His Asn Arg Gly Leu Thr Gly Ser Gly Val Lys Val Ala Val Leu Asp
20 25 30
Thr Gly Ile Ser Thr His Pro Asp Leu Asn Ile Arg Gly Gly Ala Ser
35 40 45
Phe Val Pro Gly Glu Pro Ser Thr Gln Asp Gly Asn Gly His Gly Thr
50 55 60
His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu
65 70 75 80
Gly Val Ala Pro Ser Ala Glu Leu Tyr Ala Val Lys Val Leu Gly Ala
85 90 95
Ser Gly Ser Gly Ser Val Ser Ser Ile Ala Gln Gly Leu Glu Trp Ala
100 105 110
Gly Asn Asn Gly Met His Val Ala Asn Leu Ser Leu Gly Ser Pro Ser
115 120 125
Pro Ser Ala Thr Leu Glu Gln Ala Val Asn Ser Ala Thr Ser Arg Gly
130 135 140
Val Leu Val Val Ala Ala Ser Gly Asn Ser Gly Ala Gly Ser Ile Ser
145 150 155 160
Tyr Pro Ala Arg Tyr Ala Asn Ala Met Ala Val Gly Ala Thr Asp Gln
165 170 175
Asn Asn Asn Arg Ala Ser Phe Ser Gln Tyr Gly Ala Gly Leu Asp Ile
180 185 190
Val Ala Pro Gly Val Asn Val Gln Ser Thr Tyr Pro Gly Ser Thr Tyr
195 200 205
Ala Ser Leu Asn Gly Thr Ser Met Ala Thr Pro His Val Ala Gly Ala
210 215 220
Ala Ala Leu Val Lys Gln Lys Asn Pro Ser Trp Ser Asn Val Gln Ile
225 230 235 240
Arg Asn His Leu Lys Asn Thr Ala Thr Ser Leu Gly Ser Thr Asn Leu
245 250 255
Tyr Gly Ser Gly Leu Val Asn Ala Glu Ala Ala Thr Arg
260 265
<210>293
<211>279
<212>PRT
<213>Thermoactinomyces vulgaris
<220>
<223>Thermitase S08.007
<400>293
Tyr Thr Pro Asn Asp Pro Tyr Phe Ser Ser Arg Gln Tyr Gly Pro Gln
1 5 10 15
Lys Ile Gln Ala Pro Gln Ala Trp Asp Ile Ala Glu Gly Ser Gly Ala
20 25 30
Lys Ile Ala Ile Val Asp Thr Gly Val Gln Ser Asn His Pro Asp Leu
35 40 45
Ala Gly Lys Val Val Gly Gly Trp Asp Phe Val Asp Asn Asp Ser Thr
50 55 60
Pro Gln Asn Gly Asn Gly His Gly Thr His Cys Ala Gly Ile Ala Ala
65 70 75 80
Ala Val Thr Asn Asn Ser Thr Gly Ile Ala Gly Thr Ala Pro Lys Ala
85 90 95
Ser Ile Leu Ala Val Arg Val Leu Asp Asn Ser Gly Ser Gly Thr Trp
100 105 110
Thr Ala Val Ala Asn Gly Ile Thr Tyr Ala Ala Asp Gln Gly Ala Lys
115 120 125
Val Ile Ser Leu Ser Leu Gly Gly Thr Val Gly Asn Ser Gly Leu Gln
130 135 140
Gln Ala Val Asn Tyr Ala Trp Asn Lys Gly Ser Val Val Val Ala Ala
145 150 155 160
Ala Gly Asn Ala Gly Asn Thr Ala Pro Asn Tyr Pro Ala Tyr Tyr Ser
165 170 175
Asn Ala Ile Ala Val Ala Ser Thr Asp Gln Asn Asp Asn Lys Ser Ser
180 185 190
Phe Ser Thr Tyr Gly Ser Val Val Asp Val Ala Ala Pro Gly Ser Trp
195 200 205
Ile Tyr Ser Thr Tyr Pro Thr Ser Thr Tyr Ala Ser Leu Ser Gly Thr
210 215 220
Ser Met Ala Thr Pro His Val Ala Gly Val Ala Gly Leu Leu Ala Ser
225 230 235 240
Gln Gly Arg Ser Ala Ser Asn Ile Arg Ala Ala Ile Glu Asn Thr Ala
245 250 255
Asp Lys Ile Ser Gly Thr Gly Thr Tyr Trp Ala Lys Gly Arg Val Asn
260 265 270
Ala Tyr Lys Ala Val Gln Tyr
275
<210>294
<211>401
<212>PRT
<213>Bacillus sp.AK1
<220>
<223>Thermophilic serine proteinase precursor (Ak.1
protease)S08.009
<400>294
Met Lys Phe Lys Ala Ile Val Ser Leu Ser Leu Ala Val Ser Met Ser
1 5 10 15
Leu Phe Pro Phe Leu Val Glu Ala Ala Ser Asn Asp Gly Val Glu Ser
20 25 30
Pro Lys Thr Val Ser Glu Ile Asn Val Ser His Glu Lys Gly Ala Tyr
35 40 45
Val Gln Gly Glu Val Ile Val Gln Phe Lys Glu Gln Val Asn Ala Glu
50 55 60
Glu Lys Ala Lys Ala Leu Lys Glu Val Gly Ala Thr Ala Val Pro Asp
65 70 75 80
Asn Asp Arg Val Lys Ser Lys Phe Asn Val Leu Lys Val Gly Asn Val
85 90 95
Glu Ala Val Val Lys Ala Leu Asn Asn Asn Pro Leu Val Glu Tyr Ala
100 105 110
Glu Pro Asn Tyr Leu Phe Asn Ala Ala Trp Thr Pro Asn Asp Thr Tyr
115 120 125
Tyr Gln Gly Tyr Gln Tyr Gly Pro Gln Asn Thr Tyr Thr Asp Tyr Ala
130 135 140
Trp Asp Val Thr Lys Gly Ser Ser Gly Gln Glu Ile Ala Val Ile Asp
145 150 155 160
Thr Gly Val Asp Tyr Thr His Pro Asp Leu Asp Gly Lys Val Ile Lys
165 170 175
Gly Tyr Asp Phe Val Asp Asn Asp Tyr Asp Pro Met Asp Leu Asn Asn
180 185 190
His Gly Thr His Val Ala Gly Ile Ala Ala Ala Glu Thr Asn Asn Ala
195 200 205
Thr Gly Ile Ala Gly Met Ala Pro Asn Thr Arg Ile Leu Ala Val Arg
210 215 220
Ala Leu Asp Arg Asn Gly Ser Gly Thr Leu Ser Asp Ile Ala Asp Ala
225 230 235 240
Ile Ile Tyr Ala Ala Asp Ser Gly Ala Glu Val Ile Asn Leu Ser Leu
245 250 255
Gly Cys Asp Cys His Thr Thr Thr Leu Glu Asn Ala Val Asn Tyr Ala
260 265 270
Trp Asn Lys Gly Ser Val Val Val Ala Ala Ala Gly Asn Asn Gly Ser
275 280 285
Ser Thr Thr Phe Glu Pro Ala Ser Tyr Glu Asn Val Ile Ala Val Gly
290 295 300
Ala Val Asp Gln Tyr Asp Arg Leu Ala Ser Phe Ser Asn Tyr Gly Thr
305 310 315 320
Trp Val Asp Val Val Ala Pro Gly Val Asp Ile Val Ser Thr Ile Thr
325 330 335
Gly Asn Arg Tyr Ala Tyr Met Ser Gly Thr Ser Met Ala Ser Pro His
340 345 350
Val Ala Gly Leu Ala Ala Leu Leu Ala Ser Gln Gly Arg Asn Asn Ile
355 360 365
Glu Ile Arg Gln Ala Ile Glu Gln Thr Ala Asp Lys Ile Ser Gly Thr
370 375 380
Gly Thr Tyr Phe Lys Tyr Gly Arg Ile Asn Ser Tyr Asn Ala Val Thr
385 390 395 400
Tyr
<210>295
<211>1591
<212>DNA
<213>Bacillus sp
<220>
<223>Thermophilic serine proteinase precursor(Ak.1
protease)S08.009
<400>295
gcatgctacg attaaatatc taagaatccc caattaaata ccattatttt acaaaattat 60
attatattaa aataaatgtc gagatttttg agaaaaaatt tttcattcct cccccttcct 120
caacgtaaaa ttaataccgc aattccattt aaagggggaa tatttccatg aagtttaaag 180
cgattgtaag tttatccctc gctgtttcca tgtcactttt tccattcctt gtggaagcag 240
cctctaatga tggggtagaa tctccaaaaa ccgtttccga aattaacgtg tctcatgaaa 300
aaggagcata tgtccaggga gaagtcattg tccaattcaa agaacaagta aatgctgaag 360
aaaaggcaaa ggcattaaaa gaagttgggg caacggcggt tccagataat gatcgagtta 420
aatctaaatt taatgtacta aaagtaggaa atgtggaagc tgttgtgaaa gcattaaata 480
ataacccgtt agtagaatac gcagagccaa actatttatt taatgcagct tggactccaa 540
atgatacgta ctatcagggt tatcaatatg gtccacaaaa tacgtatacc gactatgctt 600
gggatgttac aaaaggcagt agcggtcaag agattgctgt tattgataca ggtgtagatt 660
atacacatcc tgatttagat ggaaaagtca tcaaaggata tgatttcgta gataatgatt 720
acgacccaat ggatttgaat aatcatggta cgcacgtagc tggaatagca gctgcggaaa 780
caaacaatgc tacaggtatt gccggcatgg ccccaaacac aagaattttg gctgtgcgcg 840
ctttagatcg aaatggcagt ggtactctaa gtgatattgc cgatgcgatc atttatgctg 900
ctgattcagg cgctgaagtc attaacctgt cacttggttg tgattgtcat acaaccacat 960
tggagaatgc tgtaaactat gcatggaata agggttctgt agtagttgcc gcagccggaa 1020
ataatggatc ctctacaaca tttgaaccgg cttcttatga aaatgtaatt gcagttggcg 1080
cagtagatca atatgatcgg ttagcatcat tctcgaacta tggaacatgg gtagatgtcg 1140
tagctccagg tgtagacatt gtctcaacta ttactggtaa ccgatatgcc tatatgtcgg 1200
gcacttccat ggcatcccct catgtagccg gtcttgcagc cttactagcg agtcaaggac 1260
ggaataacat agaaattcgt caagccatcg agcaaacagc agacaaaatc tccggaactg 1320
gaacatactt caaatatgga agaatcaatt cttataatgc tgtaacatat taaatagatt 1380
taaataacga caccgtgcca acgccaaggg cacggtgtct taattcaagc tttttggaaa 1440
atcacgacat acaaattatg taagaaagct tattctccca taggaattga agaggaaaac 1500
caatattatt cttttatctc agcaaaaata aaaaaactgc caaatggcag actctaacta 1560
actatttttt gcttccttct ttctgctgca g 1591
<210>296
<211>1167
<212>PRT
<213>Streptococcus pyogenes
<220>
<223>C5a peptidase precursor(SCP)S08.020
<400>296
Met Arg Lys Lys Gln Lys Leu Pro Phe Asp Lys Leu Ala Ile Ala Leu
1 5 10 15
Met Ser Thr Ser Ile Leu Leu Asn Ala Gln Ser Asp Ile Lys Ala Asn
20 25 30
Thr Val Thr Glu Asp Thr Pro Val Thr Glu Gln Ala Val Glu Thr Pro
35 40 45
Gln Pro Thr Ala Val Ser Glu Glu Val Pro Ser Ser Lys Glu Thr Lys
50 55 60
Thr Pro Gln Thr Pro Asp Asp Ala Glu Glu Thr Ile Ala Asp Asp Ala
65 70 75 80
Asn Asp Leu Ala Pro Gln Ala Pro Ala Lys Thr Ala Asp Thr Pro Ala
85 90 95
Thr Ser Lys Ala Thr Ile Arg Asp Leu Asn Asp Pro Ser Gln Val Lys
100 105 110
Thr Leu Gln Glu Lys Ala Gly Lys Gly Ala Gly Thr Val Val Ala Val
115 120 125
Ile Asp Ala Gly Phe Asp Lys Asn His Glu Ala Trp Arg Leu Thr Asp
130 135 140
Lys Thr Lys Ala Arg Tyr Gln Ser Lys Glu Asp Leu Glu Lys Ala Lys
145 150 155 160
Lys Glu His Gly Ile Thr Tyr Gly Glu Trp Val Asn Asp Lys Val Ala
165 170 175
Tyr Tyr His Asp Tyr Ser Lys Asp Gly Lys Thr Ala Val Asp Gln Glu
180 185 190
His Gly Thr His Val Ser Gly Ile Leu Ser Gly Asn Ala Pro Ser Glu
195 200 205
Thr Lys Glu Pro Tyr Arg Leu Glu Gly Ala Met Pro Glu Ala Gln Leu
210 215 220
Leu Leu Met Arg Val Glu Ile Val Asn Gly Leu Ala Asp Tyr Ala Arg
225 230 235 240
Asn Tyr Ala Gln Ala Ile Arg Asp Ala Val Asn Leu Gly Ala Lys Val
245 250 255
Ile Asn Met Ser Phe Gly Asn Ala Ala Leu Ala Tyr Ala Asn Leu Pro
260 265 270
Asp Glu Thr Lys Lys Ala Phe Asp Tyr Ala Lys Ser Lys Gly Val Ser
275 280 285
Ile Val Thr Ser Ala Gly Asn Asp Ser Ser Phe Gly Gly Lys Thr Arg
290 295 300
Leu Pro Leu Ala Asp His Pro Asp Tyr Gly Val Val Gly Thr Pro Ala
305 310 315 320
Ala Ala Asp Ser Thr Leu Thr Val Ala Ser Tyr Ser Pro Asp Lys Gln
325 330 335
Leu Thr Glu Thr Ala Met Val Lys Thr Asp Asp Gln Gln Asp Lys Glu
340 345 350
Met Pro Val Leu Ser Thr Asn Arg Phe Glu Pro Asn Lys Ala Tyr Asp
355 360 365
Tyr Ala Tyr Ala Asn Arg Gly Met Lys Glu Asp Asp Phe Lys Asp Val
370 375 380
Lys Gly Lys Ile Ala Leu Ile Glu Arg Gly Asp Ile Asp Phe Lys Asp
385 390 395 400
Lys Val Ala Asn Ala Lys Lys Ala Gly Ala Val Gly Val Leu Ile Tyr
405 410 415
Asp Asn Gln Asp Lys Gly Phe Pro Ile Glu Leu Pro Asn Val Asp Gln
420 430 425
Met Pro Ala Ala Phe Ile Ser Arg Lys Asp Gly Leu Leu Leu Lys Asp
435 440 445
Asn Pro Gln Lys Thr Ile Thr Phe Asn Ala Thr Pro Lys Val Leu Pro
450 455 460
Thr Ala Ser Gly Thr Lys Leu Ser Arg Phe Ser Ser Trp Gly Leu Thr
465 470 475 480
Ala Asp Gly Asn Ile Lys Pro Asp Ile Ala Ala Pro Gly Gln Asp Ile
485 490 495
Leu Ser Ser Val Ala Asn Asn Lys Tyr Ala Lys Leu Ser Gly Thr Ser
500 505 510
Met Ser Ala Pro Leu Val Ala Gly Ile Met Gly Leu Leu Gln Lys Gln
515 520 525
Tyr Glu Thr Gln Tyr Pro Asp Met Thr Pro Ser Glu Arg Leu Asp Leu
530 535 540
Ala Lys Lys Val Leu Met Ser Ser Ala Thr Ala Leu Tyr Asp Glu Asp
545 550 555 560
Glu Lys Ala Tyr Phe Ser Pro Arg Gln Gln Gly Ala Gly Ala Val Asp
565 570 575
Ala Lys Lys Ala Ser Ala Ala Thr Met Tyr Val Thr Asp Lys Asp Asn
580 585 590
Thr Ser Ser Lys Val His Leu Asn Asn Val Ser Asp Lys Phe Glu Val
595 600 605
Thr Val Thr Val His Asn Lys Ser Asp Lys Pro Gln Glu Leu Tyr Tyr
610 615 620
Gln Ala Thr Val Gln Thr Asp Lys Val Asp Gly Lys His Phe Ala Leu
625 630 635 640
Ala Pro Lys Val Leu Tyr Glu Ala Ser Trp Gln Lys Ile Thr Ile Pro
645 650 655
Ala Asn Ser Ser Lys Gln Val Thr Val Pro Ile Asp Ala Ser Arg Phe
660 665 670
Ser Lys Asp Leu Leu Ala Gln Met Lys Asn Gly Tyr Phe Leu Glu Gly
675 680 685
Phe Val Arg Phe Lys Gln Asp Pro Thr Lys Glu Glu Leu Met Ser Ile
690 695 700
Pro Tyr Ile Gly Phe Arg Gly Asp Phe Gly Asn Leu Ser Ala Val Glu
705 710 715 720
Lys Pro Ile Tyr Asp Ser Lys Asp Gly Ser Ser Tyr Tyr His Glu Ala
725 730 735
Asn Ser Asp Ala Lys Asp Gln Leu Asp Gly Asp Gly Leu Gln Phe Tyr
740 745 750
Ala Leu Lys Asn Asn Phe Thr Ala Leu Thr Thr Glu Ser Asn Pro Trp
755 760 765
Thr Ile Ile Lys Ala Val Lys Glu Gly Val Glu Asn Ile Glu Asp Ile
770 775 780
Glu Ser Ser Glu Ile Thr Glu Thr Ile Phe Ala Gly Thr Phe Ala Lys
785 790 795 800
Gln Asp Asp Asp Ser His Tyr Tyr Ile His Arg His Ala Asn Gly Glu
805 810 815
Pro Tyr Ala Ala Ile Ser Pro Asn Gly Asp Gly Asn Arg Asp Tyr Val
820 825 830
Gln Phe Gln Gly Thr Phe Leu Arg Asn Ala Lys Asn Leu Val Ala Glu
835 840 845
Val Leu Asp Lys Glu Gly Asn Val Val Trp Thr Ser Glu Val Thr Glu
850 855 860
Gln Val Val Lys Asn Tyr Asn Asn Asp Leu Ala Ser Thr Leu Gly Ser
865 870 875 880
Thr Arg Phe Glu Lys Thr Arg Trp Asp Gly Lys Asp Lys Asp Gly Lys
885 890 895
Val Val Ala Asn Gly Thr Tyr Thr Tyr Arg Val Arg Tyr Thr Pro Ile
900 905 910
Ser Ser Gly Ala Lys Glu Gln His Thr Asp Phe Asp Val Ile Val Asp
915 920 925
Asn Thr Thr Pro Glu Val Ala Thr Ser Ala Thr Phe Ser Thr Glu Asp
930 935 940
Arg Arg Leu Thr Leu Ala Ser Lys Pro Lys Thr Ser Gln Pro Val Tyr
945 950 955 960
Arg Glu Arg Ile Ala Tyr Thr Tyr Met Asp Glu Asp Leu Pro Thr Thr
965 970 975
Glu Tyr Ile Ser Pro Asn Glu Asp Gly Thr Phe Thr Leu Pro Glu Glu
980 985 990
Ala Glu Thr Met Glu Gly Ala Thr Val Pro Leu Lys Met Ser Asp Phe
995 1000 1005
Thr Tyr Val Val Glu Asp Met Ala Gly Asn Ile Thr Tyr Thr Pro Val
1010 1015 1020
Thr Lys Leu Leu Glu Gly His Ser Asn Lys Pro Glu Gln Asp Gly Ser
1025 1030 1035 1040
Gly Gln Thr Pro Asp Lys Lys Pro Glu Ala Lys Pro Glu Gln Asp Gly
1045 1050 1055
Ser Asp Gln Ala Pro Asp Lys Lys Pro Glu Ala Lys Pro Glu Gln Asp
1060 1065 1070
Gly Ser Gly Gln Thr Pro Asp Lys Lys Pro Glu Thr Lys Pro Glu Lys
1075 1080 1085
Asp Ser Ser Gly Gln Thr Pro Gly Lys Thr Pro Gln Lys Gly Gln Pro
1090 1095 1100
Ser Arg Thr Leu Glu Lys Arg Ser Ser Lys Arg Ala Leu Ala Thr Lys
1105 1110 1115 1120
Ala Ser Thr Arg Asp Gln Leu Pro Thr Thr Asn Asp Lys Asp Thr Asn
1125 1130 1135
Arg Leu His Leu Leu Lys Leu Val Met Thr Thr Phe Phe Phe Gly Leu
1140 1145 1150
Val Ala His Ile Phe Lys Thr Lys Arg Gln Lys Glu Thr Lys Lys
1155 1160 1165
<210>297
<211>4637
<212>DNA
<213>Streptococcus pyogenes
<220>
<223>C5a peptidase precursor(SCP)S08.020
<400>297
cctgtattga tacatagccc aagaatatgc gtaaaaatag actcaatcaa gcctttttct 60
tgaaagcgac gggaataatt tttgccatag gttgtgaaat gtggcacctt gtcatccaaa 120
gtcaaaccta aaaaccagcg ataggcagta ttcacttcaa tatccttgat ggtctgacgc 180
atggaacgaa tgccataaaa gcactgaatc agagggattt taaccaacat gacaggatca 240
aggctagggc gaccattatc agacgaatag gtgtcttcta ctaggtcata gatgaagtcg 300
aagtcaatca cagcttccac ttagcgcagg aaatgttcat caggcactaa ctggtcaagg 360
gtatagaaac cgtactgatt gcggttatag tcaggttttt ctttgtgtaa cataggagaa 420
cctcacaagt cctttttctt atattatact cctaaatatc atgaaaagcc cctagaaaag 480
cgattctaag gactttgtct tcagtctgaa gagagaaccg atcggttctc tcttttatgt 540
ctaaaagaat gaggataagg aggtcacaaa ctaaacaact cttaaaaagc tgacctttac 600
taataatcgt ctttttttta taataaagat gttagtaata taattgataa atgagataca 660
tttaatcatt atggcaaaag caagaaaaat agctgtatca tatgcaaata acccctgttt 720
gctctttaaa aaagatatta tccttatttc tccacgcaca gatggacagc taggagagaa 780
tcgtttgatt ctctcttttc ttaatggtca taaagacaaa gtctctcatc aggaaaggac 840
gacacattgc gtaaaaaaca aaaattacca tttgataaac ttgccattgc gctcatgtct 900
acgagcatct tgctcaatgc acaatcagac attaaagcaa atactgtgac agaagacact 960
cctgttaccg aacaagccgt agaaacccca caaccaacag cggtttctga ggaagtacca 1020
tcatcaaagg aaaccaaaac cccacaaact cctgatgacg cagaagaaac aatagcagat 1080
gacgctaatg atctagcccc tcaagctcct gctaaaactg ctgatacacc agcaacctca 1140
aaagcgacta ttagggattt gaacgaccct tctcaggtca aaaccctgca ggaaaaagca 1200
ggcaagggag ctgggactgt tgttgcagtg attgatgctg gttttgataa aaatcatgaa 1260
gcgtggcgct taacagacaa aaccaaagca cgttaccaat caaaagaaga tcttgaaaaa 1320
gctaaaaaag agcacggtat tacctatggc gagtgggtca atgataaggt tgcttattac 1380
cacgactata gcaaagatgg taaaaccgct gtcgatcaag agcacggcac acacgtgtca 1440
gggatcttgt caggaaatgc tccatctgaa acgaaagaac cttaccgcct agaaggtgcg 1500
atgcctgagg ctcaattgct tttgatgcgt gtcgaaattg taaatggact agcagactat 1560
gctcgtaact acgctcaagc tatcagagat gctgtcaact tgggagctaa ggtgattaat 1620
atgagctttg gtaatgctgc actagcctac gctaacctcc cagacgaaac caaaaaagcc 1680
tttgactatg ccaaatcaaa aggtgttagc attgtgacct cagctggtaa tgatagtagc 1740
tttgggggca agacccgtct acctctagca gatcatcctg attatggggt ggttgggaca 1800
cctgcagcgg cagactcaac attgacagtt gcttcttaca gcccagataa acagctcact 1860
gaaactgcta tggtcaaaac agacgatcag caagataaag aaatgcctgt tctttcaaca 1920
aaccgttttg agccaaacaa ggcttacgac tatgcttatg ctaatcgtgg gatgaaagag 1980
gatgatttta aggatgtcaa aggcaagatt gcccttattg aacgtggcga tattgatttc 2040
aaagataagg ttgcaaacgc taaaaaagct ggtgctgtag gggtcttgat ctatgacaat 2100
caggacaagg gcttcccgat tgaattgcca aatgttgatc agatgcctgc ggcctttatc 2160
agtcgaaaag acggtctctt attaaaagac aatccccaaa aaaccatcac cttcaatgcg 2220
acacctaagg tattgccaac agcaagtggc accaaactaa gccgcttctc aagctggggt 2280
ctgacagctg acggcaatat taagccagat attgcagcac ccggccaaga tattttgtca 2340
tcagtggcta acaacaagta tgccaaactt tctggaacta gtatgtctgc gccattagta 2400
gcgggtatca tgggactgtt gcaaaagcaa tatgagacac agtatcctga tatgacacca 2460
tcagagcgtc ttgatttagc taaaaaagta ttgatgagct cagcaactgc cttatatgat 2520
gaagatgaaa aagcttattt ttctcctcgc caacaaggag caggagcggt cgatgctaaa 2580
aaagcttcag cagcaacgat gtatgtgaca gataaggata atacctcaag caaggttcac 2640
ctgaacaatg tttctgataa atttgaagta acagtaactg ttcacaacaa atctgataaa 2700
cctcaagagt tgtattacca agcaactgtt caaacagata aagtagatgg aaaacacttt 2760
gccttggctc ctaaagtatt gtatgaggca tcatggcaaa aaatcacaat tccagccaat 2820
agcagcaaac aagtcaccgt tccaatcgat gctagtcgat ttagcaagga cttgcttgcc 2880
caaatgaaaa atggctattt cttagaaggt tttgttcgtt tcaaacaaga tcctacaaaa 2940
gaagagctta tgagcattcc atatattggt ttccgaggtg attttggcaa tctgtcagcc 3000
gtagaaaaac caatctatga tagcaaagac ggtagcagct actatcatga agcaaatagt 3060
gatgccaaag accaattaga tggtgatgga ttacagtttt acgctctgaa aaacaacttt 3120
acagcactta ccacagagtc taacccatgg acgattatta aggctgtcaa agaaggggtt 3180
gaaaacatag aggatatcga atcttcagag atcacagaaa ccatttttgc aggtactttt 3240
gcaaaacaag acgatgatag ccactactat atccaccgtc acgctaatgg cgagccatat 3300
gctgcgatct ctccaaatgg ggacggtaac agagattatg tccaattcca aggtactttc 3360
ttgcgtaatg ctaaaaacct tgtggctgaa gtcttggaca aagaaggaaa tgttgtttgg 3420
acaagtgagg taaccgagca agttgttaaa aactacaaca atgacttggc aagcacactt 3480
ggttcaaccc gttttgaaaa aactcgttgg gacggtaaag ataaagacgg caaagttgtt 3540
gctaacggaa cctacaccta tcgtgtccgc tacactccga ttagttcagg tgcaaaagaa 3600
caacacactg attttgatgt gattgtagac aatacgacac ctgaagtcgc aacatcggca 3660
acattctcaa cagaagatcg tcgtttgaca cttgcatcta aaccaaaaac cagccaaccg 3720
gtttaccgtg agcgtattgc ttacacttat atggatgagg atctgccaac aacagagtat 3780
atttctccaa atgaagatgg tacctttact cttcctgaag aggctgaaac aatggaaggc 3840
gctactgttc cattgaaaat gtcagacttt acttatgttg ttgaagatat ggctggtaat 3900
atcacttata caccagtgac taagctattg gagggccact ctaataagcc agaacaagac 3960
ggttcaggtc aaacaccaga taaaaaacca gaagctaaac cagaacaaga cggttcagat 4020
caagcaccag acaaaaaacc agaagctaaa ccagaacaag acggttcagg tcaaacacca 4080
gataaaaaac cagaaactaa accagaaaaa gatagttcag gtcaaacacc aggtaaaact 4140
cctcaaaaag gtcaaccttc tcgtactcta gagaaacgct cttctaagcg tgctttagct 4200
acaaaagcat caacaagaga tcagttacca acgactaatg acaaggatac aaatcgttta 4260
catctcctta agttagttat gaccactttc ttctttggat tagtagctca tatctttaaa 4320
acaaaacgcc aaaaagaaac taaaaaatag tattacttaa actgataaga gcttatttaa 4380
tagagggaac caatttggtt ctctctattt ttttagatgt gctgcctcat caaaaagaag 4440
ttaggacaaa tagctgtcaa caattgcctt ttgcttatct tttcagttgc tgtgataaaa 4500
gatggcacgc caaacgcatg aagaaacttt tgttttcaga aaactagatt cataatgaca 4560
aaatcaatca aagtagtaca ctatagatga ggtgactacg atgattgatt ttattatttc 4620
tcaaccattc tatttct 4637
<210>298
<211>699
<212>PRT
<213>Fervidobacterium pennivorans
<220>
<223>fervidolysin S08.020
<400>298
Met Arg Lys Val Leu Leu Ile Ala Ser Ile Val Ala Leu Ile Leu Ala
1 5 10 15
Leu Phe Ser Cys Ala Asn Pro Ser Phe Glu Pro Arg Ser Lys Ala Lys
20 25 30
Asp Leu Ala Ser Leu Pro Glu Ile Lys Ser Gln Gly Tyr His Ile Leu
35 40 45
Phe Gly Glu Leu Arg Asp Gly Glu Tyr Thr Glu Gly Lys Ile Leu Val
50 55 60
Gly Tyr Asn Asp Arg Ser Glu Val Asp Lys Ile ValLys Ala Val Asn
65 70 75 80
Gly Lys Val Val Leu Glu Leu Pro Gln Ile Lys Val ValSer Ile Lys
85 90 95
Leu Asn Gly Met Thr Val Lys Gln Ala Tyr Asp Lys Ile Lys Ala Leu
100 105 110
Ala Leu Lys Gly Ile Arg Tyr Val Glu Pro Ser Tyr Lys Arg Glu Leu
115 120 125
Ile Lys Pro Thr Val Val Lys Pro Asn Pro Asp Met Tyr Lys Ile Arg
130 135 140
Lys Pro Gly Leu Asn Ser Thr Ala Arg Asp Tyr Gly Glu Glu Leu Ser
145 150 155 160
Asn Glu Leu Trp Gly Leu Glu Ala Ile Gly Val Thr Gln Gln Leu Trp
165 170 175
Glu Glu Ala Ser Gly Thr Asn Ile Ile Val Ala Val Val Asp Thr Gly
180 185 190
Val Asp Gly Thr His Pro Asp Leu Glu Gly Gln Val Ile Ala Gly Tyr
195 200 205
Arg Pro Ala Phe Asp Glu Glu Leu Pro Ala Gly Thr Asp Ser Ser Tyr
210 215 220
Gly Gly Ser His Gly Thr His Val Ala Gly Thr Ile Ala Ala Lys Lys
225 230 235 240
Asp Gly Lys Gly Ile Val Gly Val Ala Pro Gly Ala Lys Ile Met Pro
245 250 255
Ile Val Ile Phe Asp Asp Pro Ala Leu Val Gly Gly Asn Gly Tyr Val
260 265 270
Gly Asp Asp Tyr Val Ala Ala Gly Ile Ile Trp Ala Thr Asp His Gly
275 280 285
Ala Lys Val Met Asn His Ser Trp Gly Gly Trp Gly Tyr Ser Tyr Thr
290 295 300
Met Lys Glu Ala Phe Asp Tyr Ala Met Glu His Gly Val Val Met Val
305 310 315 320
Val Ser Ala Gly Asn Asn Thr Ser Asp Ser His His Gln Tyr Pro Ala
325 330 335
Gly Tyr Pro Gly Val Ile Gln Val Ala Ala Leu Asp Tyr Tyr Gly Gly
340 345 350
Thr Phe Arg Val Ala Gly Phe Ser Ser Arg Ser Asp Gly Val Ser Val
355 360 365
Gly Ala Pro Gly Val Thr Ile Leu Ser Thr Val Pro Gly Glu Asp Ser
370 375 380
Ile Gly Tyr Glu Gly His Asn Glu Asn Val Pro Ala Thr Asn Gly Gly
385 390 395 400
Thr Tyr Asp Tyr Tyr Gln Gly Thr Ser Met Ala Ala Pro His Val Thr
405 410 415
Gly Val Val Ala Val Leu Leu Gln Lys Phe Pro Asn Ala Lys Pro Trp
420 425 430
Gln Ile Arg Lys Leu Leu Glu Asn Thr Ala Phe Asp Phe Asn Gly Asn
435 440 445
Gly Trp Asp His Asp Thr Gly Tyr Gly Leu Val Lys Leu Asp Ala Ala
450 455 460
Leu Gln Gly Pro Leu Pro Thr Gln Gly Gly Val Glu Glu Phe Gln Val
465 470 475 480
Val Val Thr Asp Ala Lys Gly Asn Phe Gly Val Pro Thr Val Phe Val
485 490 495
Ser Met Met Arg Asp Asn Gly Ser Cys Tyr Tyr Ala Lys Thr Gly Pro
500 505 510
Asp Gly Ile Ala Arg Phe Pro His Ile Asp Ser Gly Thr Tyr Asp Ile
515 520 525
Phe Val Gly Gly Pro Asp His Trp Asp Arg Ala Leu Ala Pro Tyr Asp
530 535 540
Gly Glu Ser Ile Pro Gly Gly Tyr Ala Ile Ala Leu Arg Met Ala Glu
545 550 555 560
Glu Arg Gln Ala Ser Phe Val Gly Phe Gly Val Ser Pro Asp Ala Thr
565 570 575
Gln Leu Asn Val Asn Phe Asn Ser Thr Leu Gln Val Lys Phe Ser Thr
580 585 590
Asn Leu Ser Thr Leu Lys Asp Pro Gln Phe Val Val Val Asp Pro Leu
595 600 605
Leu Arg Gly Val Tyr Gly Arg Val Ala Tyr Ala Arg Asn Gln Thr Tyr
610 615 620
Asp Leu Ser Leu Leu Ser Gly Gln Ile Ser Phe Gly Ile Gln Thr Leu
625 630 635 640
Leu Pro Ala Ala Thr Asp Ile Thr Ile Gln Gly Thr Val Thr Leu Asn
645 650 655
Gly Glu Asp Ile Pro Val Tyr Gly Val Leu Lys Ala Gly Thr Thr Trp
660 665 670
Thr Ile Ile Asp Asp Phe Gly Gly Leu Asn Leu Gly Thr Asp Ser Gln
675 680 685
Pro Ile Tyr Val Trp Trp Thr Ile Phe Gly Gln
690 695
<210>299
<211>2227
<212>DNA
<213>Fervidobacterium pennivorans
<220>
<223>fervidolysin S08.020
<400>299
aaaacagaga tttaccacag atttgtaacc ggtgagaact aagaaagggg ggattgcgat 60
gagaaaagta ttgttgatag cttccattgt tgctcttatt ctcgcgctct tttcatgtgc 120
aaatccgagt tttgagccaa ggagcaaagc aaaagattta gcatcgttac cggagataaa 180
atctcagggt taccatatac tcttcggtga attgagggac ggggaatata ccgagggtaa 240
aatcctcgtt ggttacaacg ataggtcgga agtagataaa atagtcaaag cagttaacgg 300
taaagtagtg ctcgaattac cacagattaa agtggtctcc ataaaactca atggaatgac 360
ggttaaacaa gcatacgaca agataaaggc acttgcactt aaaggtatca ggtatgttga 420
accaagttac aagagggagc tcataaaacc aacagttgtg aagccaaatc ctgatatgta 480
caaaatcaga aaacctgggt tgaacagtac tgcaagagat tacggtgaag aactttccaa 540
cgaactctgg ggccttgagg caattggtgt aacacaacag ctttgggaag aagcatcggg 600
aacaaatatt atcgttgcgg ttgttgatac tggtgtcgac ggcacccatc cagatttgga 660
agggcaagtt atcgcaggat acagaccggc atttgatgaa gaactcccag caggaacaga 720
ttcatcctac ggtggttcac atggaacaca cgttgctgga acgattgcgg caaagaaaga 780
tggaaaaggt atcgttggtg ttgcacctgg tgcaaagata atgccgatag ttatattcga 840
tgacccagca cttgttggtg gaaatggtta cgttggcgac gattatgtcg cagcaggtat 900
aatttgggca acagaccatg gtgcaaaggt aatgaaccac tcttggggtg gctggggtta 960
cagctacaca atgaaagaag catttgatta tgcgatggag catggagttg tcatggttgt 1020
ctcagctggt aacaatacat cagattcaca ccatcaatat ccagcaggtt atcctggtgt 1080
tatccaagtt gctgctcttg actactacgg tggaacattt agagtggcag gtttttcaag 1140
cagaagcgat ggagtctcag ttggcgcacc tggagttacg attctctcga cggttcctgg 1200
cgaagatagt atcggatacg aagggcacaa cgaaaacgtt cctgcaacaa atggcggtac 1260
ttatgattac tatcaaggaa cctcaatggc tgcaccgcat gtaacaggtg ttgttgctgt 1320
attgctacaa aaattcccga acgcaaaacc atggcagata agaaaattac tcgaaaacac 1380
agcattcgat tttaacggaa atggttggga tcacgatacc ggttatggtc tggtaaagct 1440
cgatgcagca ttgcaaggtc ctcttccgac acaaggtgga gtcgaagagt tccaagtagt 1500
tgttacggat gctaaaggaa atttcggagt cccaacggtc tttgtatcaa tgatgagaga 1560
caatggttca tgctattatg cgaaaacagg tcctgatggt atagcaaggt tcccacacat 1620
cgacagcggc acttacgaca tatttgttgg tggaccagac cactgggata gagctctggc 1680
tccttacgat ggagaaagta ttcccggagg ttatgccatt gctttgagaa tggctgagga 1740
aaggcaagct tccttcgtcg gctttggagt ttctccagat gcaacacaac tcaatgtcaa 1800
ttttaattcg acactgcaag ttaaattcag cactaacctc tcaacactga aagacccaca 1860
atttgtggtt gttgatccgt tactaagagg agtttacggt cgtgtggcat acgcaagaaa 1920
tcaaacatac gatttgtcgt tactctcagg tcaaatttca tttggtatac aaacgctctt 1980
accagcagcg acagatatta caatccaagg aacagtaaca ttgaacggtg aagacatccc 2040
tgtttacgga gttctaaaag ctggaaccac atggactata attgatgact ttggtggatt 2100
gaatttgggc accgactcgc agccaatcta tgtctggtgg acaatcttcg gacagtgata 2160
gtttcatttt aacgtcaaag caaaccgatg gggtgttcag accccatcgg ttttttgttt 2220
tgccttt 2227
<210>300
<211>381
<212>PRT
<213>Geobacillus stearothermophilus
<220>
<223>Subtilisin J precursor S08.035
<400>300
Met Arg Ser Lys Lys Leu Trp Ile Ser Leu Leu Phe Ala Leu Thr Leu
1 5 10 15
Ile Phe Thr Met Ala Phe Ser Asn Met Ser Val Gln Ala Ala Gly Lys
20 25 30
Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys Gln Thr Met Ser
35 40 45
Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser Glu Lys Gly Gly
50 55 60
Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala Ala Ala Thr Leu
65 70 75 80
Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro Ser Val Ala Tyr
85 90 95
ValGlu Glu Asp His Ile Ala His Glu Tyr Ala Gln Ser Val Pro Tyr
100 105 110
Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser Gln Gly Tyr Thr
115 120 125
Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly Ile Asp Ser Ser
130 135 140
His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe Val Pro Ser Glu
145 150 155 160
Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr His Val Ala Gly
165 170 175
Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu Gly Val Ser Pro
180 185 190
Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser Thr Gly Ser Gly
195 200 205
Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala Ile Ser Asn Asn
210 215 220
Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Ser Gly Ser Thr Ala
225 230 235 240
Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly Ile Val Val Ala
245 250 255
Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly Ser Ser Ser Thr Val Gly
260 265 270
Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala Val Gly Ala Val Asn Ser
275 280 285
Ser Asn Gln Arg Ala Ser Phe Ser Ser Ala Gly Ser Glu Leu Asp Val
290 295 300
Met Ala Pro Gly Val Ser Ile Gln Ser Thr Leu Pro Gly Gly Thr Tyr
305 310 315 320
Gly Ala Tyr Asn Gly Thr Ser Met Ala Thr Pro His Val Ala Gly Ala
325 330 335
Ala Ala Leu Ile Leu Ser Lys His Pro Thr Trp Thr Asn Ala Gln Val
340 345 350
Arg Asp Arg Leu Glu Ser Thr Ala Thr Tyr Leu Gly Asn Ser Phe Tyr
355 360 365
Tyr Gly Lys Gly Leu Ile Asn Val Gln Ala Ala Ala Gln
370 375 380
<210>301
<211>2494
<212>DNA
<213>Geobacillus stearothermophilus
<220>
<223>Subtilisin J precursor S08.035
<400>301
gaattctcca ttttcttctg ctatcaaaat aacagactcg tgattttcca aacgagcttt 60
caaaaaagcc tctgcccctt gcaaatcgga tgcctgtcta taaaattccc gatattggtt 120
aaacaacggc gcaatggcgg ccgcatctga tgtctttgct tggcgaatgt tcatcttatt 180
tcttcctccc tctcaataat tttttcattc tatccctttt ctgtaaagtt tatttttcag 240
aatactttta tcatcatgct ttgaaaaaat atcacgataa tatccattgt tctcacggaa 300
gcacacgcag gtcatttgaa cgaatttttt cgacagaaat ttgccgggac tcaggagcat 360
ttaacctaaa aaagcatgac atttcagcat aatgaacatt tactcatgtc tattttcgtt 420
cttttctgta tgaaaatagt tatttcgagt ctctacggaa atagcgagag atgatatacc 480
taaatagaga taaaatcatc tcaaaaaaat gggtctacta aaatattatt ccatctatta 540
caataaattc acagaatagt cttttaagta agtctactct gaattttttt aaaaggagag 600
ggtaaagagt gagaagcaaa aaattgtgga tcagcttgtt gtttgcgtta acgttaatct 660
ttacgatggc gttcagcaac atgtctgtgc aggctgccgg aaaaagcagt acagaaaaga 720
aatacattgt cggatttaaa cagacaatga gtgccatgag ttccgccaag aaaaaggatg 780
ttatttctga aaaaggcgga aaggttcaaa agcaatttaa gtatgttaac gcggccgcag 840
caacattgga tgaaaaagct gtaaaagaat tgaaaaaaga tccgagcgtt gcatatgtgg 900
aagaagatca tattgcacat gaatatgcgc aatctgttcc ttatggcatt tctcaaatta 960
aagcgccggc tcttcactct caaggctaca caggctctaa cgtaaaagta gctgttatcg 1020
acagcggaat tgactcttct catcctgact taaacgtcag aggcggagca agcttcgtac 1080
cttctgaaac aaacccatac caggacggca gttctcacgg tacgcatgta gccggtacga 1140
ttgccgctct taataactca atcggtgttc tgggcgtcag cccaagcgca tcattatatg 1200
cagtaaaagt gcttgattca acaggaagcg gccaatatag ctggattatt aacggcattg 1260
aatgggccat ttccaacaat atggatgtta ttaacatgag ccttggcgga ccttctggtt 1320
ctacagcgct gaaaacagtc gttgataaag ccgtttccag cggtatcgtc gttgctgccg 1380
ctgccggaaa cgaaggttcg tccggaagct caagcacagt cggctaccct gcaaaatatc 1440
cttctactat tgcggtaggt gcggtaaaca gcagcaacca aagagcttca ttctcaagcg 1500
caggttctga gcttgatgtg atggctcctg gcgtatccat ccaaagcaca cttcctggag 1560
gcacttacgg tgcttacaac ggcacgtcca tggcgactcc tcacgttgcc ggagcagcag 1620
cgctaattct ttctaagcac ccgacttgga caaacgcgca agtccgtgat cgtttagaaa 1680
gcactgcaac atatcttgga aactctttct actatggaaa agggttaatc aacgtacaag 1740
cagctgcaca ataatagtaa aaagaagcag gttcctccat acctgcttct ttttatttgt 1800
cagcatcctg atgttccggc gcattctctt ctttctccgt atgttgaatc cgttccatga 1860
tcgaaggatg gctgcctctg aaaatcttca caagcaccgg aggatcaacc tggctcagcc 1920
ccgtcacggc caaatcctga aacgttttaa cagcggcttc tctgttctct gtcaactcga 1980
tcccatactg gtcagcctta ttctcctgat aacgcgagac agcattagaa aaaggcgtca 2040
ccgcaaagct caaaacagaa aacaaaagca ataacagcgg aagtgccgca agatcatgcc 2100
gcccttctaa atgaaacatg ctgcgggtta ggcgaaccgt ccgcttgtaa agcttatcaa 2160
tgacataaaa tccggcgagc gacacgagca aataaccagc cagaccgatg taaacgtgct 2220
tcatgacata atggcccatt tcgtggccca taataaacag gatttctgaa tcgtcaagtt 2280
tgttcagcgt cgtatcccac aatacaatcc gtttattggc cccaattcct gtaacatagg 2340
cattcagcgc atttgttttt tctgacatgt tcacttcata tacatggtca gccggaatat 2400
tggcttcatc tgccagctct aaaattttgc tttcaagctc tttgtttttc agcggataaa 2460
aatcattgta taaaggatcg actctagagg atcc 2494
<210>302
<211>381
<212>PRT
<213>Bacillus subtilis
<220>
<223>Subtilisin E precursor S08.036
<400>302
Met Arg Ser Lys Lys Leu Trp Ile Ser Leu Leu Phe Ala Leu Thr Leu
1 5 10 15
Ile Phe Thr Met Ala Phe Ser Asn Met Ser Val Gln Ala Ala Gly Lys
20 25 30
Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys Gln Thr Met Ser
35 40 45
Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser Glu Lys Gly Gly
50 55 60
Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala Ala Ala Thr Leu
65 70 75 80
Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro Ser Val Ala Tyr
85 90 95
Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln Ser Val Pro Tyr
100 105 110
Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser Gln Gly Tyr Thr
115 120 125
Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly Ile Asp Ser Ser
130 135 140
His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe Val Pro Ser Glu
145 150 155 160
Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr His Val Ala Gly
165 170 175
Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu Gly Val Ser Pro
180 185 190
Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser Thr Gly Ser Gly
195 200 205
Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala Ile Ser Asn Asn
210 215 220
Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr Gly Ser Thr Ala
225 230 235 240
Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly Ile Val Val Ala
245 250 255
Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly Ser Thr Ser Thr Val Gly
260 265 270
Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala Val Gly Ala Val Asn Ser
275 280 285
Ser Asn Gln Arg Ala Ser Phe Ser Ser Ala Gly Ser Glu Leu Asp Val
290 295 300
Met Ala Pro Gly Val Ser Ile Gln Ser Thr Leu Pro Gly Gly Thr Tyr
305 310 315 320
Gly Ala Tyr Asn Gly Thr Ser Met Ala Thr Pro His Val Ala Gly Ala
325 330 335
Ala Ala Leu Ile Leu Ser Lys His Pro Thr Trp Thr Asn Ala Gln Val
340 345 350
Arg Asp Arg Leu Glu Ser Thr Ala Thr Tyr Leu Gly Asn Ser Phe Tyr
355 360 365
Tyr Gly Lys Gly Leu Ile Asn Val Gln Ala Ala Ala Gln
370 375 380
<210>303
<211>1500
<212>DNA
<213>Bacillus subtilis
<220>
<223>Subtilisin E precursor S08.036
<400>303
gatataccta aatagagata aaatcatctc aaaaaaatgg gtctactaaa atattattcc 60
atctattaca ataaattcac agaatagtct tttaagtaag tctactctga atttttttaa 120
aaggagaggg taaagagtga gaagcaaaaa attgtggatc agcttgttgt ttgcgttaac 180
gttaatcttt acgatggcgt tcagcaacat gtctgcgcag gctgccggaa aaagcagtac 240
agaaaagaaa tacattgtcg gatttaaaca gacaatgagt gccatgagtt ccgccaagaa 300
aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag caatttaagt atgttaacgc 360
ggccgcagca acattggatg aaaaagctgt aaaagaattg aaaaaagatc cgagcgttgc 420
atatgtggaa gaagatcata ttgcacatga atatgcgcaa tctgttcctt atggcatttc 480
tcaaattaaa gcgccggctc ttcactctca aggctacaca ggctctaacg taaaagtagc 540
tgttatcgac agcggaattg actcttctca tcctgactta aacgtcagag gcggagcaag 600
cttcgtacct tctgaaacaa acccatacca ggacggcagt tctcacggta cgcatgtagc 660
cggtacgatt gccgctctta ataactcaat cggtgttctg ggcgttagcc caagcgcatc 720
attatatgca gtaaaagtgc ttgattcaac aggaagcggc caatatagct ggattattaa 780
cggcattgag tgggccattt ccaacaatat ggatgttatc aacatgagcc ttggcggacc 840
tactggttct acagcgctga aaacagtcgt tgacaaagcc gtttccagcg gtatcgtcgt 900
tgctgccgca gccggaaacg aaggttcatc cggaagcaca agcacagtcg gctaccctgc 960
aaaatatcct tctactattg cagtaggtgc ggtaaacagc agcaaccaaa gagcttcatt 1020
ctccagcgca ggttctgagc ttgatgtgat ggctcctggc gtgtccatcc aaagcacact 1080
tcctggaggc acttacggcg cttataacgg aacgtccatg gcgactcctc acgttgccgg 1140
agcagcagcg ttaattcttt ctaagcaccc gacttggaca aacgcgcaag tccgtgatcg 1200
tttagaaagc actgcaacat atcttggaaa ctctttctac tatggaaaag ggttaatcaa 1260
cgtacaagca gctgcacaat aatagtaaaa agaagcaggt tcctccatac ctgcttcttt 1320
ttatttgtca gcatcctgat gttccggcgc attctcttct ttctccgcat gttgaatccg 1380
ttccatgatc gacggatggc tgcctctgaa aatcttcaca agcaccggag gatcaacctg 1440
ctcagccccg tcacggccaa atcctgaaac gttttaacac tggcttctct gttctctgtc 1500
<210>304
<211>274
<212>PRT
<213>Bacillus licheniformis
<220>
<223>Subtilisin DY S08.037
<400>304
Ala Gln Thr Val Pro Tyr Gly Ile Pro Leu Ile Lys Ala Asp Lys Val
1 5 10 15
Gln Ala Gln Gly Tyr Lys Gly Ala Asn Val Lys Val Gly Ile Ile Asp
20 25 30
Thr Gly Ile Ala Ala Ser His Thr Asp Leu Lys Val Val Gly Gly Ala
35 40 45
Ser Phe Val Ser Gly Glu Ser Tyr Asn Thr Asp Gly Asn Gly His Gly
50 55 60
Thr His Val Ala Gly Thr Val Ala Ala Leu Asp Asn Thr Thr Gly Val
65 70 75 80
Leu Gly Val Ala Pro Asn Val Ser Leu Tyr Ala Ile Lys Val Leu Asn
85 90 95
Ser Ser Gly Ser Gly Thr Tyr Ser Ala Ile Val Ser Gly Ile Glu Trp
100 105 110
Ala Thr Gln Asn Gly Leu Asp Val Ile Asn Met Ser Leu Gly Gly Pro
115 120 125
Ser Gly Ser Thr Ala Leu Lys Gln Ala Val Asp Lys Ala Tyr Ala Ser
130 135 140
Gly Ile Val Val Val Ala Ala Ala Gly Asn Ser Gly Ser Ser Gly Ser
145 150 155 160
Gln Asn Thr Ile Gly Tyr Pro Ala Lys Tyr Asp Ser Val Ile Ala Val
165 170 175
Gly Ala Val Asp Ser Asn Lys Asn Arg Ala Ser Phe Ser Ser Val Gly
180 185 190
Ala Glu Leu Glu Val Met Ala Pro Gly Val Ser Val Tyr Ser Thr Tyr
195 200 205
Pro Ser Asn Thr Tyr Thr Ser Leu Asn Gly Thr Ser Met Ala Ser Pro
210 215 220
His Val Ala Gly Ala Ala Ala Leu Ile Leu Ser Lys Tyr Pro Thr Leu
225 230 235 240
Ser Ala Ser Gln Val Arg Asn Arg Leu Ser Ser Thr Ala Thr Asn Leu
245 250 255
Gly Asp Ser Phe Tyr Tyr Gly Lys Gly Leu Ile Asn Val Glu Ala Ala
260 265 270
Ala Gln
<210>305
<211>384
<212>PRT
<213>Engyodontium album
<220>
<223>Proteinase K precursor S08.054
<400>305
Met Arg Leu Ser Val Leu Leu Ser Leu Leu Pro Leu Ala Leu Gly Ala
1 5 10 15
Pro Ala Val Glu Gln Arg Ser Glu Ala Ala Pro Leu Ile Glu Ala Arg
20 25 30
Gly Glu Met Val Ala Asn Lys Tyr Ile Val Lys Phe Lys Glu Gly Ser
35 40 45
Ala Leu Ser Ala Leu Asp Ala Ala Met Glu Lys Ile Ser Gly Lys Pro
50 55 60
Asp His Val Tyr Lys Asn Val Phe Ser Gly Phe Ala Ala Thr Leu Asp
65 70 75 80
Glu Asn Met Val Arg Val Leu Arg Ala His Pro Asp Val Glu Tyr Ile
85 90 95
Glu Gln Asp Ala Val Val Thr Ile Asn Ala Ala Gln Thr Asn Ala Pro
100 105 110
Trp Gly Leu Ala Arg Ile Ser Ser Thr Ser Pro Gly Thr Ser Thr Tyr
115 120 125
Tyr Tyr Asp Glu Ser Ala Gly Gln Gly Ser Cys Val Tyr Val Ile Asp
130 135 140
Thr Gly Ile Glu Ala Ser His Pro Glu Phe Glu Gly Arg Ala Gln Met
145 150 155 160
Val Lys Thr Tyr Tyr Tyr Ser Ser Arg Asp Gly Asn Gly His Gly Thr
165 170 175
His Cys Ala Gly Thr Val Gly Ser Arg Thr Tyr Gly Val Ala Lys Lys
180 185 190
Thr Gln Leu Phe Gly Val Lys Val Leu Asp Asp Asn Gly Ser Gly Gln
195 200 205
Tyr Ser Thr Ile Ile Ala Gly Met Asp Phe Val Ala Ser Asp Lys Asn
210 215 220
Asn Arg Asn Cys Pro Lys Gly Val Val Ala Ser Leu Ser Leu Gly Gly
225 230 235 240
Gly Tyr Ser Ser Ser Val Asn Ser Ala Ala Ala Arg Leu Gln Ser Ser
245 250 255
Gly Val Met Val Ala Val Ala Ala Gly Asn Asn Asn Ala Asp Ala Arg
260 265 270
Asn Tyr Ser Pro Ala Ser Glu Pro Ser Val Cys Thr Val Gly Ala Ser
275 280 285
Asp Arg Tyr Asp Arg Arg Ser Ser Phe Ser Asn Tyr Gly Ser Val Leu
290 295 300
Asp Ile Phe Gly Pro Gly Thr Ser Ile Leu Ser Thr Trp Ile Gly Gly
305 310 315 320
Ser Thr Arg Ser Ile Ser Gly Thr Ser Met Ala Thr Pro His Val Ala
325 330 335
Gly Leu Ala Ala Tyr Leu Met Thr Leu Gly Lys Thr Thr Ala Ala Ser
340 345 350
Ala Cys Arg Tyr Ile Ala Asp Thr Ala Asn Lys Gly Asp Leu Ser Asn
355 360 365
Ile Pro Phe Gly Thr Val Asn Leu Leu Ala Tyr Asn Asn Tyr Gln Ala
370 375 380
<210>306
<211>2347
<212>DNA
<213>Engyodontium album
<220>
<223>Proteinase K precursor S08.054
<400>306
cgaacagtat aaagaaatag aggggaccta aactcatgtt caactctagg gccccgttga 60
atcattcccc caaagtggga acgccagtgg cgacaaatct gtatactgtt tcctgctttt 120
tctggtttgt caaggttctc attaccctct tggctttctc ctcttttcgc gacattgtgc 180
ttgtggtgtg cctatgtatg ttgtagtcaa tgctatatgt caatggtttt tgtcaaatgc 240
catggcctag tggctctatg ctggagctgc taattctggg cctttatcga ctcctccgac 300
tagtcctgat gaagaagttg caacactacc agcgtcacat tcgccccatt gatcacttct 360
tgatccttcg caataacctt cacggctgct cgaatgactt gacgtcacga tgggcctact 420
atccaacttc cagaacagga aagtcgagat agttctacat agaccatgga cgagttcatg 480
agatgttgga ttcaaggata aatagctggc catgtcgccg cttcgataca gtttgtttca 540
tctccatcat cagcatcaat cgcagcaata caaagcatcc tctcaaactc aacaaacttt 600
cttgattaag caccgctttc tcattcatca acaacagcca aaatgcgttt gtctgttctt 660
ctgagtcttc ttcccctcgc tctcggcgct cctgccgttg agcagcgctc cgaggctgct 720
cctctgatcg aggcccgcgg cgagatggtt gccaacaagt acattgtcaa gttcaaggag 780
ggtagcgctc tttctgctct cgatgctgcc atggagaaga tttctggcaa gcccgaccac 840
gtctacaaga acgtcttcag tggtttcgct gcgacccttg acgagaacat ggttcgggtt 900
ctccgcgccc atcccgatgt aagttttccc atctataatc tcttccccta aaaagtctct 960
cccttctaat atcgccgtca ggttgagtac attgagcagg atgctgttgt caccatcaac 1020
gctgcgcaga ccaacgctcc ctggggcctt gctcgcatct ccagcaccag ccccggtacc 1080
tctacttact actatgacga atctgccggc caaggctcct gcgtctacgt gattgacacc 1140
ggtatcgagg catcgcaccc cgagtttgag ggtcgtgccc agatggtcaa gacctactac 1200
tactccagtc gcgacggtaa cggtcacggc actcactgcg ctggtaccgt tggctcccga 1260
acctacggtg tcgccaagaa gacccagctc tttggtgtca aggtcctcga tgacaacggc 1320
agtggccagt actccaccat catcgccggt atggactttg ttgccagcga caagaacaac 1380
cgcaactgcc ccaaaggtgt cgttgcctcc ttgtcccttg gcggtggtta ctcctcctcc 1440
gtgaacagcg ccgctgccag gctccagagc tctggtgtca tggtcgccgt cgctgccggt 1500
aacaacaacg ctgacgcccg caactactcc cctgcttctg agccctcggt ctgcactgtc 1560
ggtgcttctg accgctacga cagacgctcc agcttctcca actacggcag cgttttggac 1620
atctttggcc ctggtaccag cattctctcc acctggatcg gcggcagcac ccgctccatc 1680
tctggaactt ccatggctac tccccacgtt gccggtctcg ctgcctacct catgactctt 1740
ggaaagacta ccgccgccag cgcttgccga tacattgccg acaccgccaa caagggcgac 1800
ttgagcaaca ttcccttcgg cactgtcaac ctgcttgcct acaacaacta ccaggcttaa 1860
gtccttcagt caacgccaaa agctcaagat atgaaacaag ttatgaatgc atctgtatat 1920
agacgagcat acttcatatc agtacataac catccttagt atatttaaaa aggcaacgaa 1980
tttgcagatt tagctgtcag acagttagat ttgcatgaat cagaagatat ttgctttcat 2040
aaattgcctc gtctcctttt tgcgcttcac ttattatctt tcttgacaaa ttgaaacagt 2100
ggcattaatt tcctttctaa ggaaatcgca tttaaatagt gaatgtaatc atatttgcac 2160
ttactagagg taggaccaaa ttatcgagga acagttggtc atcgggcggg ttcctcggct 2220
tctttctgtt ttagcaagtg tgtagtcctc gtgcatggcc gatgatgatt agtctgagag 2280
aacttgtgag ctaacaacgg catctcgagg aaacgttcaa ctatgagccg aaagacaagt 2340
acttaat 2347
<210>307
<211>534
<212>PRT
<213>Vibrio alginolyticus
<220>
<223>Alkaline serine exoprotease A precursor S08.050
<400>307
Met Leu Lys Lys Leu Leu Ser Cys Cys Ile Thr Ser Ala Leu Cys Phe
1 5 10 15
His Ser Ser Leu Ala Phe Ser Gln Pro Asn Glu Ile Ala Asp Ser Ala
20 25 30
Glu Leu Gln Gln Ala Pro Asp Thr Leu Pro Ala Thr Leu Met Leu Ala
35 40 45
Pro Asp Asp Ile Ala Ile Ala Asp Arg Tyr Ile Val Val Phe Gln Gln
50 55 60
Pro Gln Met Met Ala Ser Ser Ser Pro Glu Phe Glu Gln Phe Thr Gln
65 70 75 80
Gln Ser Val Asp Arg Met Ser Gly Leu Tyr Ser Ile Gln Val Glu Ser
85 90 95
Val Phe Asp His Ser Ile Ser Gly Phe Val Ala Asn Leu Ser Pro Glu
100 105 110
Gln Leu Lys Asp Leu Arg Ser Asp Pro Arg Val Asp Tyr Ile Glu Gln
115 120 125
Asp Arg Ile Leu Ser Leu Asp Pro Ile Val Ser Ala Asp Ala Asn Gln
130 135 140
Thr Asn Ala Ile Trp Gly Leu Asp Arg Ile Asp Gln Arg Asn Leu Pro
145 150 155 160
Leu Asp Asn Asn Tyr Ser Ala Asn Phe Asp Gly Thr Gly Val Thr Ala
165 170 175
Tyr Val Ile Asp Thr Gly Val Asn Asn Ala His Val Glu Phe Gly Gly
180 185 190
Arg Ser Val Ser Gly Tyr Asp Phe Val Asp Asn Asp Ala Asp Ala Ser
195 200 205
Asp Cys Asn Gly His Gly Thr His Val Ala Gly Thr Ile Gly Gly Ser
210 215 220
Leu Tyr Gly Val Ala Lys Asn Val Asn Leu Val Gly Val Arg Val Leu
225 230 235 240
Ser Cys Ser Gly Ser Gly Ser Thr Ser Gly Val Ile Ala Gly Val Asp
245 250 255
Trp Val Ala Ala Asn Ala Ser Gly Pro Ser Val Ala Asn Met Ser Leu
260 265 270
Gly Gly Gly Gln Ser Val Ala Leu Asp Ser Ala Val Gln Ser Ala Val
275 280 285
Gln Ser Gly Val Ser Phe Met Leu Ala Ala Gly Asn Ser Asn Ala Asp
290 295 300
Ala Cys Asn Tyr Ser Pro Ala Arg ValAla Thr Gly Val Thr Val Gly
305 310 315 320
Ser Thr Thr Ser Thr Asp Ala Arg Ser Ser Phe Ser Asn Trp Gly Ser
325 330 335
Cys Val Asp Val Phe Ala Pro Gly Ser Gln Ile Lys Ser Ala Trp Tyr
340 345 350
Asp Gly Gly Tyr Lys Thr Ile Ser Gly Thr Ser Met Ala Thr Pro His
355 360 365
Val Ala Gly Val Ala Ala Leu Tyr Leu Gln Glu Asn Ser Ser Val Ser
370 375 380
Pro Ser Gln Val Glu Ala Leu Ile Val Ser Arg Ala Ser Thr Gly Lys
385 390 395 400
Val Thr Asp Thr Arg Gly Ser Val Asn Lys Leu Leu Tyr Ser Leu Thr
405 410 415
Asp Ala Asp Cys Gly Gln Asp Cys Gly Gly Pro Asp Pro Thr Pro Asp
420 425 430
Pro Glu Gly Lys Leu Thr Ser Gly Val Pro Val Ser Gly Leu Ser Gly
435 440 445
Ser Ser Gly Gln Val Ala Tyr Tyr Tyr Val Asp Val Glu Ala Gly Gln
450 455 460
Arg Leu Thr Val Gln Met Tyr Gly Gly Ser Gly Asp Ala Asp Leu Tyr
465 470 475 480
Leu Arg Phe Gly Ala Lys Pro Thr Leu Asn Ala Trp Asp Cys Arg Pro
485 490 495
Phe Lys Tyr Gly Asn Asn Glu Thr Cys Thr Val Ser Ala Thr Gln Ser
500 505 510
Gly Arg Tyr His Val Met Ile Gln Gly Tyr Ser Asn Tyr Ser Gly Val
515 520 525
Ser Ile Gln Ala Asn Tyr
530
<210>308
<211>1856
<212>DNA
<213>Vibrio alginolyticus
<220>
<223>Alkaline serine exoprotease A precursor S08.050
<400>308
ctgcagcacg tattgagcat accacgctga ttgtctgtgt tttcgtgttg acagtttggt 60
actaggtaaa atcaacaaga ctatgatttc aaggaacatt ttatgttaaa gaaactacta 120
agttgttgta tcacctccgc gctctgtttt cattcttctc tcgctttttc gcaacccaat 180
gagattgcgg acagtgcaga gttacaacaa gctcccgaca cattgcctgc cactttgatg 240
cttgcaccgg acgacatcgc cattgcagat cgatatatag tggtatttca acaaccgcaa 300
atgatggcga gcagctcacc ggaattcgag caattcacgc aacagtctgt agaccgcatg 360
tccggtttat attcgataca agtggagtcg gtttttgacc actcgatcag tggatttgtc 420
gctaacttga gtcctgagca actaaaagat ctgcgttctg atcctcgtgt ggactacatt 480
gagcaagaca gaatcctatc gcttgaccca atagtctcgg cagacgcaaa tcaaaccaat 540
gccatttggg gactagatcg aatcgaccaa cgtaacttgc cactcgataa caactacagt 600
gccaactttg atgggactgg tgtaacggct tatgttatcg atactggtgt gaacaatgca 660
catgttgagt ttggtgggcg ctcggtttct gggtatgact ttgtcgataa tgatgcagat 720
gcaagtgact gtaatggaca cggcacacac gtggcgggca ccattggcgg cagcttgtat 780
ggtgttgcga aaaacgtcaa ccttgtcggc gtgagagtat tgagctgtag cggatcgggg 840
tctacgtctg gtgttatcgc cggtgtggat tgggtggctg cgaacgcttc cggaccttca 900
gttgccaata tgagtttagg tggcggtcaa tctgtcgctc tcgatagtgc ggtgcaaagt 960
gcggttcaat caggtgtcag ctttatgctt gcagcaggta actccaatgc cgatgcgtgt 1020
aactactctc cagcgcgcgt tgctactggt gtaactgtcg gctcgaccac cagcacggat 1080
gcacgttcga gtttttcaaa ctggggcagt tgtgtggacg tgttcgcgcc aggctcacaa 1140
atcaaatctg cgtggtatga cggtggttac aaaaccatta gtggtacatc gatggcgacg 1200
ccacatgtag cgggtgtagc agcactgtat cttcaagaaa acagttccgt gtcgccaagc 1260
caagtagagg ccttgatcgt gagccgcgca agtaccggaa aggtgacgga cacaagaggc 1320
agcgtgaaca agctacttta tagcttaacg gatgcagatt gtggccaaga ctgcggtggc 1380
ccagatccaa caccggaccc agaaggcaag ttaacctcgg gcgtgccagt gagcggttta 1440
agtggctcaa gcggtcaagt agcgtattac tatgttgatg tagaagctgg gcagcgctta 1500
accgtacaaa tgtatggtgg cagcggtgat gcggatttgt atctccgttt tggtgcaaaa 1560
ccgacactga atgcatggga ctgccgacct ttcaaatacg gtaacaatga aacatgtacg 1620
gtcagcgcga cacaaagtgg acgctaccac gtcatgattc aaggttactc aaactatagc 1680
ggtgtcagca ttcaagctaa ctactagttg ttattctcgt tactaaaaga gaacaaagcc 1740
cgcttttcgg tgggctttgc tttcgggaat actaagttta ataggtggcg ttcgatgcta 1800
ttcaaacgca ccggcctttt tcttcaagaa ctcaactcga agcggcatcg aagctt 1856
<210>309
<211>461
<212>PRT
<213>Staphylococcus epidermidis
<220>
<223>Epidermin leader peptide-processing serine
protease epiP precursor S08.060
<400>309
Met Asn Lys Phe Lys Phe Phe Ile Val Phe Leu Ile Leu Ser Leu Val
1 5 10 15
Phe Leu Gln Asn Glu Tyr Ala Phe Gly Ser Ser Leu Asn Glu Glu Leu
20 25 30
Ser Tyr Tyr Ser Val Glu Tyr Asp Asn Ala Lys Thr Phe Lys Glu Ser
35 40 45
Ile Lys Gln Lys Asn Ile Glu Leu Thr Tyr Lys Ile Pro Glu Leu His
50 55 60
Thr Ala Gln Ile Lys Thr Ser Lys Ser Lys Leu Asn Ser Leu Ile Lys
65 70 75 80
Ser Asn Lys Asn Val Lys Phe Val Asn Pro Thr Cys Ser Thr Cys Val
85 90 95
Val Glu Lys Ser Val Lys Thr Gly Lys Asn Leu Asn Asn Lys Lys Asn
100 105 110
Gly Ser His Asp Leu Phe Asp Arg Gln Trp Asp Met Arg Lys Ile Thr
115 120 125
Asn Glu Gly Lys Ser Tyr Lys Leu Ser Pro Asp Arg Lys Lys Ala Lys
130 135 140
Val Ala Leu Val Asp Ser Gly Val Asn Ser Ser His Thr Asp Leu Lys
145 150 155 160
Ser Ile Asn Lys Ile Val Asn Glu Val Pro Lys Asn Gly Phe Arg Gly
165 170 175
Ser Glu Asn Asp Glu Ser Gly Asn Lys Asn Phe Glu Glu Asp Lys Leu
180 185 190
Asn His Gly Thr Leu Val Ala Gly Gln Ile Gly Ala Asn Gly Asn Leu
195 200 205
Lys Gly Val Asn Pro Gly Val Glu Met Asn Val Tyr Arg Val Phe Gly
210 215 220
Ser Lys Lys Ser Glu Met Leu Trp Val Ser Lys Gly Ile Ile Asp Ala
225 230 235 240
Ala Asn Asp Asp Asn Asp Val Ile Asn Val Ser Leu Gly Asn Tyr Leu
245 250 255
Ile Lys Asp Asn Gln Asn Lys Lys Lys Leu Arg Asp Asp Glu Lys Val
260 265 270
Asp Tyr Asp Ala Leu Gln Lys Ala Ile Asn Tyr Ala Gln Lys Lys Gly
275 280 285
Ser Ile Val Val Ala Ala Val Gly Asn Asp Gly Ile Asn Val Lys Lys
290 295 300
Val Lys Glu Ile Asn Lys Lys Arg Asn Leu Asn Ser Lys Thr Ser Lys
305 310 315 320
Lys Val Tyr Asp Ser Pro Ala Asn Leu Asn Asn Val Met Thr Val Gly
325 330 335
Ser Ile Asp Asp Asn Asp Tyr Ile Ser Glu Phe Ser Asn Tyr Gly Asn
340 345 350
Asn Phe Ile Asp Leu Met Thr Ile Gly Gly Ser Tyr Lys Leu Leu Asp
355 360 365
Lys Tyr Gly Lys Asp Ala Trp Leu Glu Lys Gly Tyr Met Gln Lys Gln
370 375 380
Ser Val Leu Ser Thr Ser Ser Asn Gly Arg Tyr Ile Tyr Gln Ser Gly
385 390 395 400
Thr Ser Leu Ala Ala Pro Lys Val Ser Gly Ala Leu Ala Leu Glu Ile
405 410 415
Asp Lys Tyr Gln Leu Lys Asp Gln Pro Glu Thr Ala Ile Glu Leu Phe
420 425 430
Lys Lys Lys Gly Ile Glu Lys Glu Lys Tyr Met Asp Lys Lys His Tyr
435 440 445
Gly Asn Gly Lys Leu Asp Val Tyr Lys Leu Leu Lys Glu
450 455 460
<210>310
<211>8700
<212>DNA
<213>Staphylococcus epidermidis
<220>
<223>Epidermin leader peptide-processing serine
protease epiP precursor S08.060
<400>310
agatcttgtg ttatataact aaacaaattt ctccattcgt atttagaaaa ttgactttta 60
tcaagtttat ccaaatatat atttccagta tattctgtat ttaacccagc taatatattt 120
aataatgtac tttttccaca cccactttca cctataatat tgtagatata acctttatga 180
agatccaaac ttatagaatt tattatttgt ttattgtctt ttgtgaagtt caaatcattt 240
atttccattt tttgaacaaa gttattgtaa gttgttttaa tagttaatac ctcttctggt 300
tctttattta tttttaaaat tctatctgaa gatccaattg ctcgttgtac ttccgtccaa 360
taagatgtaa tagatactat tggattaata atttgaaata aatataaaac ataagcaaac 420
atatctccgc ttttcatcat attattttcc attaagtaat aacccaaaaa taaaatacca 480
aaaatgttaa taaatagaat taagttcata attggttcga aaaaagataa tactttgatc 540
ttatgtaact ctatatcgaa tatatttttt aatagggtat agttttttat tttttcgata 600
ttatatgtac ttaaagtttt tattaatttt attgtagata atctattact ataataagaa 660
gataatttag cagtagcttc ttgagattta cttgatactc ttttcattat atttcctata 720
ggtagtatta caattatcaa tataggtaat gtacacacta aatataatgt caaggttttg 780
ttaattatat ataaaaatat tagtgatact ataactgaaa ataaattcta cagaaaaaac 840
tctagttatg ttcatagtat cgtttactaa cctactagtt aagttacttg ctgagttttt 900
taagtgaaaa ctataaggta actttatcac tttattccat gtaacacttc taatgttttg 960
tattattttt tgacctatat atccaagaat ataagtagaa acaccagaaa atattaaagt 1020
cagaccaaaa catataataa tgattacaat tttatctgtt gataagctag atttgtttaa 1080
ggcatttcta attattaaag gaatgtataa tgaaaaacta gttccaatca aactaaatat 1140
tagtccaata cttaaaagta gagtgttagg tttggttatt ttccataaat catatagacc 1200
tttgataata tcatcacctt ttaaacttta tatcattaat ataatgttta ggaaaagtag 1260
aagaaaatta cacttttgta attttctgaa tatacatagt atttattttg ggggagtact 1320
aaaataataa ttgaaaaggg ttttataatc ctttttaata aatttttagg agtgtttaaa 1380
atggaagcag taaaagaaaa aaatgatctt tttaatcttg atgttaaagt taatgcaaaa 1440
gaatctaacg attcaggagc tgaaccaaga attgctagta aatttatatg tactcctgga 1500
tgtgcaaaaa caggtagttt taacagttat tgttgttaat tcagaagaat tagattggca 1560
gggcttcaat agaggctctg tcttaatttt gaggtgaaat agaattggat aatatatttg 1620
ttccatcgaa tatatatatg gtaagaactc ctatattttc aattgaatta tataatcaat 1680
tcttaaaatc tgacaatata gattatgact taattttaca aaacgatatt tttaaagaat 1740
ctataatgac aacgacatat aatctttatc aaagtattgg caaaatagac tgggaaaagg 1800
ataataaaaa aaccagaaat gtaaaagaaa gtttattaaa atatctcata agaatgagta 1860
ctagaagtac accatatgga atgctaagcg gtgtagcttt aggggaattt agtgaaaata 1920
ataatattaa aattaaggac tcttcgtttc ataaaaaaga tgtaaaaata gatgggcaat 1980
ggttatataa attagtccat tatttagaaa gcgattacac atattataaa gacagttttg 2040
tcatatggaa tcaacaaaat tatatttata acaatcgttt atatttagat aataattcat 2100
caatcactga aaataaaaga aatgatgtat tatctgtcaa atacaattct atattagtgt 2160
ttatacatga gaattctaaa aaaaatatta cttatgaaga acttgtacaa ttgatatcta 2220
gtaagtacag tatagaaaat aaagaagaag taaaagtatt tgttcaagaa ctcataaata 2280
aagaaattat attttctgat ttgagaccta cattagagaa taaaaatcct ttagattaca 2340
ttattaatag tttaaatcca aaaaatagtt tagttggaac acttattaat atttctaatg 2400
aaattacaaa atattctaaa atgcctttag gaaaaggaga atataaatat ttagatattg 2460
ttaatttaat gtcacaatta tttgtttcta aaaactattt gcaaatagat acctatatag 2520
attattcaag aaatgaatta aaacaaagtt tagctgataa tattagtgaa gcagcatata 2580
ttctctggtt attatctcct aatcattttg gtacaaaaac tattaggaat tatcacgaat 2640
tttttatgga taaatatgga tttgaacaac tagtaaattt aaagcaattg ctctcagata 2700
taaatggatt tggctatccc aaaaaagaca gttatagttt ttctaataac attgcatttt 2760
taaaagaaaa gtatttgctt gcaattcaaa ataacagcca tattgaaata acagaaaacg 2820
acgttaaaaa tttagaaaag aataatacag tttctaaaat caatgcgcct gtttcaactg 2880
aaatatatag tgagatatat tttggaaatt caataaaagg ttatgaggat tttgccgtga 2940
taagtccaat attaggatct tttaatgccg gtgcaacttt tggaaggttt acgggaaatt 3000
tcaatataaa gaaaaaaaat caattacaaa aagaaatagt gcatcattac aataattaca 3060
tgaatgaaaa tggtttagaa ataagccaat taaatgaagg tcctcttaac tcaagaaatg 3120
taaatatttt gaataataat agaatatata atacttgttt aaatttaaat ttacctaaaa 3180
gtgatataga tataaatgac atatttattg gagctacatt taacaaactt tatctatatt 3240
ctgaaaaaca tgattcaaga attgtattcg tatctaattc aatgtttaat tatgagtttg 3300
gatctgaatt atacaaattt ttaagagaaa tttcatttga aaaaacaaaa tttatacaac 3360
ctataactga agaaggcatt gactcattac ctttttgtcc aagaattatt tataaaaata 3420
ttattttaaa accagctact tggaaaataa attcagaaat gttttctgaa actgaaaatt 3480
ggttaaatag gttcgcaact attagagaaa aatggcatat tccaaaagat gtaattattg 3540
cttttggaga taatcgattg ctattaaatt tattaaatga caagcatctc attatactaa 3600
aaaaagaact aaaaaaacat ggtaggattc gaatattaga aagctttatc aatgaatcta 3660
ataatgagag aatgttagaa attgttacgc cattatataa aaaaactagt ttaaaagaac 3720
aatctttcat tatacctaaa aatagaaata agcacttcaa taatcttaaa gattggtttt 3780
caattcattt aagtattcct aaaacatacc aagataattt tattcaagat tatctattac 3840
catttataac ggaattaaaa gttaataatt ttattaataa atttttttac ataaaattta 3900
aagaagatga agattttata aaattaagat tattaagaga agatgaagat tattctcaaa 3960
tttattcttt cataaaaaat tggaaagatt attgcttatt aaatagtgaa ttatatgact 4020
attctatagt tgattatgtt cctgaagtat atagatatgg tggtccacac gtaattgaag 4080
atattgagaa tttttttatg tatgatagtc tattatcaat aaatataata caatcagagt 4140
tcaaaattcc aaaagaattt atcgttgcta tatcaataga ttttttatta gattatttag 4200
aaattaataa aagtgagaaa gaagaaattt taattaataa tgcggaagat ttatatcgta 4260
gtaatgacat aagagaatat aaaaatttat tagctaaact taccaatcct aaaaatgact 4320
atgaaatttt aaaaaaagaa tttccgaatc ttcatgaatt tctatttaat aaaattagta 4380
ttttagaaaa tcttaaaaag acactacaaa aaagcttata tacttcacgt tctaggataa 4440
ttggcagttt tatacacatg cgttgtaata gaatattcgg tattaatcct gaaaaagaaa 4500
aatttgtttt atctattttt aatgaaatta caaaaactaa aaaatattgg gatggttgtg 4560
attaatatta ataacattaa aaaaatttta gaaaataaaa tcaccttttt gtctgacatt 4620
gaaaaagcta catatattat agaaaatcaa agtgagtatt gggatcctta tactctatct 4680
catggttatc caggtataat acttttttta agcgcatcag aaaaagtatt tcataaagat 4740
ttagaaaaag taatacatca atatattaga aaactaggcc cttatttaga aagtggtatt 4800
gatggatttt cactttttag tggtctttcc ggaattggat ttgcgctaga cattgcgtct 4860
gataaacagt actcttatca aagtatctta gaacaaattg ataatttact tgttcaatat 4920
gtttttgatt ttttaaataa cgatgcattg gaagtaaccc ctactaacta tgatataata 4980
caaggatttt ctggtatagg aaggtacttg ttaaatagaa tatcgtataa ttataatgca 5040
aaaaaagcat taaagcatat acttaattac ttcaaaacaa ttcattactc taaagacaat 5100
tggttagttt caaatgaaca tcaattttta gatatagata agcaaaattt tccgtcagga 5160
aatataaatt taggattagc gcatggtatt ttaggtcctc tatcattaac agctttgagt 5220
aaaatgaatg ggattgaaat cgaaggccat gaagagtttt tacaagactt cacttcattt 5280
ttgctcaaac ctgaattcaa aaataataat gaatggttcg atcgctatga tatattagaa 5340
aattatatac ctaattattc cgtcagaaac ggttggtgtt acggtgatac agggattatg 5400
aatacattac ttttgtctgg taaagcctta aataatgaag gcttaattaa aatgtctaaa 5460
aatattttaa ttaacataat agataagaat aatgatgatt taatcagtcc aaccttctgt 5520
cacggactag catcgcactt aaccattatt catcaagcga ataaattctt taatctatct 5580
caagtaagca catatatcga taccattgtc agaaaaatta ttagtcatta ttctgaagaa 5640
agtagtttta tgttccaaga catagagtac tcatacggac aaaaaattta taaaaacaaa 5700
gtgggaattc tagagggtga attaggtgtt cttttagctt tactagatta tattgataca 5760
caaaaccaat caaggaaaaa ttggaaaaat atgtttttaa taacataata ggaggaataa 5820
gatatgtatg gaaaattatt gatatgcgct acagcatcga taaatgtaat taatattaat 5880
cactacatag ttgagttaaa gcaacatttt gatgaagtta atatattatt tagtcctagt 5940
agtaaaaatt ttataaatac tgatgttctc aagttatttt gtgataactt gtacgatgaa 6000
attaaagatc ctcttttaaa tcatatcaat attgtagaaa atcatgaata tattttagta 6060
ttacctgcat cagcaaatac tattaataaa atagctaatg gtatatgtga taatctttta 6120
actactgtat gtttaaccgg atatcaaaaa ttatttatat ttccaaatat gaacataaga 6180
atgtggggaa atccattttt acaaaaaaat attgatttac ttaaaaataa tgatgtgaaa 6240
gtgtattccc ctgatatgaa taaatcattc gaaatatcta gtggccgtta caaaaacaat 6300
atcacaatgc ctaatattga aaatgtacta aattttgtat taaataacga aaaaagacct 6360
ttggattaac aaaggtcttt tctaattaaa attttatatc cgagtttacg ttcattaata 6420
atttctatct ctttacaatt ttttaaacta tcccttaatc gatggatata tacatttatt 6480
gtattagaat caacaaagtc ttctgtatcc cacactccct tttttaattc ctcttttgat 6540
acatatcttc caagattaat atataagcac cgtagaattt ttaattctat attagaaaga 6600
ttaactaagt aattattaaa cacaaattga tggttttcaa agtctataaa atcatcatta 6660
acatatttaa tatacttttt tatttcattt aaaattctac ataatattaa acttttgctt 6720
tcattatttt ttataatata taaatctatg cctaaactat aaaaataaca cttcctacta 6780
tagctagtat tacctgttat tataactatt ggaatttttc ctataaattc ttttaaaaac 6840
gtataatact catcaaactt ttcatacaca attataaaat ttgggtctat atttgaagaa 6900
ttaattgtaa ttcttctatc taattctaaa atactttcaa taagaataga atctacctca 6960
ccgacaatat taatagaaat cattttattc ccttcattct ttaagtaatt tgtatacgtc 7020
tagttttcca ttaccataat gttttttatc catatatttt tctttttcta tccctttttt 7080
cttaaataac tctatagctg tttcgggttg gtcttttaat tgatacttat caatttctag 7140
tgctaaagct ccagaaacct tgggtgcagc aagtgatgtc cctgattgat atatgtatct 7200
tccattagaa gaagtactta aaacactttg tttttgcata tatccttttt ctaaccaagc 7260
atcttttcca tacttatcta aaagtttata agaacctcct atcgtcatta aatctataaa 7320
attatttcca taattagaaa actcagaaat ataatcatta tcatcgatgg atcctacagt 7380
cataacatta tttagatttg ctgggctatc atataccttt tttgatgttt tagaatttag 7440
atttcttttt ttatttattt cttttacttt ttttacattg ataccgtcat tacccacagc 7500
tgcaacaaca atactacctt ttttttgagc atagtttata gctttctgta gtgcatcgta 7560
atcaactttt tcatcatctc ttaatttttt tttattttga ttatctttaa ttaaataatt 7620
tcctaaacta acgttgatta catcattgtc atcatttgct gcatcaataa ttcctttaga 7680
tacccaaagc atttcacttt tctttgagcc aaatactcgg tatacattca tctctactcc 7740
agggtttaca ccttttaaat taccgtttgc tcctatttgt cctgctacta atgtaccatg 7800
attcaattta tcttcttcaa aatttttatt tcctgattca tcgttttcgc tacctctaaa 7860
accattttta ggcacttcat taactatctt atttatactc tttaaatctg tatgactact 7920
attcacacca gaatctacta aagcaacttt tgcttttttt ctatctggac ttagcttata 7980
acttttacct tcatttgtta tttttcgcat atcccattgt ctgtcaaata aatcatggct 8040
gccatttttt ttattattta aattttttcc tgtctttaca gatttttcaa ctacacaagt 8100
ggaacaggta ggatttacaa acttgacgtt tttattactc tttattagtg aatttaattt 8160
tgatttgcta gttttaattt gtgctgtatg tagttcagga attttataag ttaactcgat 8220
atttttttgt ttaatggatt ctttaaaagt ttttgcatta tcatattcaa cactataata 8280
acttaattct tcatttagtg aacttccaaa agcatactca ttttgcaaaa aaactaatga 8340
caatattaaa aaaacaatga aaaatttaaa tttgttcata tagcacctct aacatattat 8400
ttatattaaa cattaattta acacttatgt ttttactttt ttatttatat tatctttaat 8460
aatgttctgt tgcaagatga aaaatacgag gtatcaaagt accgatacag cgagtattac 8520
actcaattaa ttaaaaataa aatatgttgt gattaaaatt tattttataa aagtatgggc 8580
aatttattat tattcaagtt aaaacaaaga gtccgggaca taaagtttca gcctcttcgt 8640
cctaattacc aaaaaactta ctccaaaatc cttttttaga ttggtttttt ctaatttttt 8700
<210>311
<211>1052
<212>PRT
<213>Cricetulus griseus
<220>
<223>Site-1 protease(Subtilisin/kexin-isozyme 1)
S08.063
<400>311
Met Lys Leu Ile Asn Ile Trp Leu Leu Leu Leu Val Val Leu Leu Cys
1 5 10 15
Gly Lys Lys His Leu Gly Asp Arg Leu Gly Lys Lys Ala Phe Glu Lys
20 25 30
Ala Ser Cys Pro Ser Cys Ser His Leu Thr Leu Lys Val Glu Phe Ser
35 40 45
Ser Thr Val Val Glu Tyr Glu Tyr Ile Val Ala Phe Asn Gly Tyr Phe
50 55 60
Thr Ala Lys Ala Arg Asn Ser Phe Ile Ser Ser Ala Leu Lys Ser Ser
65 70 75 80
Glu Val Asp Asn Trp Arg Ile Ile Pro Arg Asn Asn Pro Ser Ser Asp
85 90 95
Tyr Pro Ser Asp Phe Glu Val Ile Gln Ile Lys Glu Lys Gln Lys Ala
100 105 110
Gly Leu Leu Thr Leu Glu Asp His Pro Asn Ile Lys Arg Val Thr Pro
115 120 125
Gln Arg Lys Val Phe Arg Ser Leu Lys Phe Ala Glu Ser Asp Pro Ile
130 135 140
Val Pro Cys Asn Glu Thr Arg Trp Ser Gln Lys Trp Gln Ser Ser Arg
145 150 155 160
Pro Leu Arg Arg Ala Ser Leu Ser Leu Gly Ser Gly Phe Trp His Ala
165 170 175
Thr Gly Arg His Ser Ser Arg Arg Leu Leu Arg Ala Ile Pro Arg Gln
180 185 190
ValAla Gln Thr Leu Gln Ala Asp Val Leu Trp Gln Met Gly Tyr Thr
195 200 205
Gly Ala Asn Val Arg Val Ala Val Phe Asp Thr Gly Leu Ser Glu Lys
210 215 220
His Pro His Phe Lys Asn Val Lys Glu Arg Thr Asn Trp Thr Asn Glu
225 230 235 240
Arg Thr Leu Asp Asp Gly Leu Gly His Gly Thr Phe Val Ala Gly Val
245 250 255
Ile Ala Ser Met Arg Glu Cys Gln Gly Phe Ala Pro Asp Ala Glu Leu
260 265 270
His Ile Phe Arg Val Phe Thr Asn Asn Gln Val Ser Tyr Thr Ser Trp
275 280 285
Phe Leu Asp Ala Phe Asn Tyr Ala Ile Leu Lys Lys Ile Asp Val Leu
290 295 300
Asn Leu Ser Ile Gly Gly Pro Asp Phe Met Asp His Pro Phe Val Asp
305 310 315 320
Lys Val Trp Glu Leu Thr Ala Asn Asn Val Ile Met Val Ser Ala Ile
325 330 335
Gly Asn Asp Gly Pro Leu Tyr Gly Thr Leu Asn Asn Pro Ala Asp Gln
340 345 350
Met Asp Val Ile Gly Val Gly Gly Ile Asp Phe Glu Asp Asn Ile Ala
355 360 365
Arg Phe Ser Ser Arg Gly Met Thr Thr Trp Glu Leu Pro Gly Gly Tyr
370 375 380
Gly Arg Val Lys Pro Asp Ile Val Thr Tyr Gly Ala Gly Val Arg Gly
385 390 395 400
Ser Gly Val Lys Gly Gly Cys Arg Ala Leu Ser Gly Thr Ser Val Ala
405 410 415
Ser Pro Val Val Ala Gly Ala Val Thr Leu Leu Val Ser Thr Val Gln
420 425 430
Lys Arg Glu Leu Val Asn Pro Ala Ser Val Lys Gln Ala Leu Ile Ala
435 440 445
Ser Ala Arg Arg Leu Pro Gly Val Asn Met Phe Glu Gln Gly His Gly
450 455 460
Lys Leu Asp Leu Leu Arg Ala Tyr Gln Ile Leu Ser Ser Tyr Lys Pro
465 470 475 480
Gln Ala Ser Leu Ser Pro Ser Tyr Ile Asp Leu Thr Glu Cys Pro Tyr
485 490 495
Met Trp Pro Tyr Cys Ser Gln Pro Ile Tyr Tyr Gly Gly Met Pro Thr
500 505 510
Ile Val Asn Val Thr Ile Leu Asn Gly Met Gly Val Thr Gly Arg Ile
515 520 525
Val Asp Lys Pro Glu Trp Arg Pro Tyr Leu Pro Gln Asn Gly Asp Asn
530 535 540
Ile Glu Val Ala Phe Ser Tyr Ser Ser Val Leu Trp Pro Trp Ser Gly
545 550 555 560
Tyr Leu Ala Ile Ser Ile Ser Val Thr Lys Lys Ala Ala Ser Trp Glu
565 570 575
Gly Ile Ala Gln Gly His Ile Met Ile Thr Val Ala Ser Pro Ala Glu
580 585 590
Thr Glu Ala Lys Asn Gly Ala Glu His Thr Ser Thr Val Lys Leu Pro
595 600 605
Ile Lys Val Lys Ile Ile Pro Thr Pro Pro Arg Ser Lys Arg Val Leu
610 615 620
Trp Asp Gln Tyr His Asn Leu Arg Tyr Pro Pro Gly Tyr Phe Pro Arg
625 630 635 640
Asp Asn Leu Arg Met Lys Asn Asp Pro Leu Asp Trp Asn Gly Asp His
645 650 655
Val His Thr Asn Phe Arg Asp Met Tyr Gln His Leu Arg Ser Met Gly
660 665 670
Tyr Phe Val Glu Val Leu Gly Ala Pro Phe Thr Cys Phe Asp Ala Thr
675 680 685
Gln Tyr Gly Thr Leu Leu Met Val Asp Ser Glu Glu Glu Tyr Phe Pro
690 695 700
Glu Glu Ile Ala Lys Leu Arg Arg Asp Val Asp Asn Gly Leu Ser Leu
705 710 715 720
Val Ile Phe Ser Asp Trp Tyr Asn Thr Ser Val Met Arg Lys Val Lys
725 730 735
Phe Tyr Asp Glu Asn Thr Arg Gln Trp Trp Met Pro Asp Thr Gly Gly
740 745 750
Ala Asn Ile Pro Ala Leu Asn Glu Leu Leu Ser Val Trp Asn Met Gly
755 760 765
Phe Ser Asp Gly Leu Tyr Glu Gly Glu Phe Ala Leu Ala Asn His Asp
770 775 780
Met Tyr Tyr Ala Ser Gly Cys Ser Ile Ala Lys Phe Pro Glu Asp Gly
785 790 795 800
Val Val Ile Thr Gln Thr Phe Lys Asp Gln Gly Leu Glu Val Leu Lys
805 810 815
Gln Glu Thr Ala Val Val Glu Asn Val Pro Ile Leu Gly Leu Tyr Gln
820 825 830
Ile Pro Ala Glu Gly Gly Gly Arg Ile Val Leu Tyr Gly Asp Ser Asn
835 840 845
Cys Leu Asp Asp Ser His Arg Gln Lys Asp Cys Phe Trp Leu Leu Asp
850 855 860
Ala Leu Leu Gln Tyr Thr Ser Tyr Gly Val Asn Pro Pro Ser Leu Ser
865 870 875 880
His Ser Gly Asn Arg Gln Arg Pro Pro Ser Gly Ala Gly Leu Ala Pro
885 890 895
Pro Glu Arg Met Glu Gly Asn His Leu His Arg Tyr Ser Lys Val Leu
900 905 910
Glu Ala His Leu Gly Asp Pro Lys Pro Arg Pro Leu Pro Ala Cys Pro
915 920 925
His Leu Ser Trp Ala Lys Pro Gln Pro Leu Asn Glu Thr Ala Pro Ser
930 935 940
Asn Leu Trp Lys His Gln Lys Leu Leu Ser Ile Asp Leu Asp Lys Val
945 950 955 960
Val Leu Pro Asn Phe Arg Ser Asn Arg Pro Gln Val Arg Pro Leu Ser
965 970 975
Pro Gly Glu Ser Gly Ala Trp Asp Ile Pro Gly Gly Ile Met Pro Gly
980 985 990
Arg Tyr Asn Gln Glu Val Gly Gln Thr Ile Pro Val Phe Ala Phe Leu
995 1000 1005
Gly Ala Met Val Ala Leu Ala Phe Phe Val Val Gln Ile Ser Lys Ala
1010 1015 1020
Lys Ser Arg Pro Lys Arg Arg Arg Pro Arg Ala Lys Arg Pro Gln Leu
1025 1030 1035 1040
Thr Gln Gln Thr His Pro Pro Arg Thr Pro Ser Val
1045 1050
<210>312
<211>4198
<212>DNA
<213>Cricetulus griseus
<220>
<223>Site-1 protease (Subtilisin/kexin-isozyme
1) S08.063
<400>312
tgttcgcggc agaggcgccg ttcccccggg cccgccgacc tcgagcctga ggcggacgca 60
ggtcggccct cagagtggtt tcttgggcat ccccactaga tttgggtctg tggtgcaaat 120
ggagtctagg actcagtcga ctctgcccta atgagagaag cccctgtcca agatggagaa 180
gaagcggaga aagaaatgaa agcctctttt tgggccaagc tgtgggtgac catgggactg 240
aggttttctt tacgttggac aagtctgtag gatggctgat cagtaaggtt gcagctttta 300
gccaaaacag aaattcactt ctgatcaagg aagaacctag tgcgatttga atttatgcaa 360
ttttatgacc atattcactt aggaccatga agctcatcaa catctggctt cttctgctgg 420
tggttttgct ctgtggaaag aagcatctgg gtgacaggct ggggaagaaa gcgtttgaaa 480
aggcatcatg ccctagctgt tcccacctga ctttgaaggt ggaattctcc tcaactgtgg 540
tggaatatga atatattgtg gctttcaacg gatacttcac agccaaagct agaaactcat 600
ttatttcaag tgctctgaaa agcagtgaag tagacaactg gagaattata cctcggaaca 660
acccatccag tgactaccct agtgattttg aggtgattca gataaaagag aagcagaagg 720
ccgggctgct cacacttgaa gatcatccaa acatcaagcg ggtgacacct caacgcaaag 780
tctttcgttc cttgaagttt gctgaatctg accccattgt gccatgtaat gaaactcggt 840
ggagccagaa gtggcagtca tcacgacccc tgagaagagc cagtctctcc ctgggctctg 900
gattctggca tgcaacagga agacattcaa gccggcgatt gctgagagcc attcctcgac 960
aggttgccca gacattgcag gcagatgtgc tgtggcagat gggatacaca ggtgctaatg 1020
tcagggttgc tgtttttgat actgggctca gtgagaagca tccacacttc aagaatgtga 1080
aggagagaac caactggacc aatgagcgga ccctggatga tgggctgggc catggcacat 1140
ttgtcgcagg tgtgattgcc agcatgaggg agtgccaggg atttgcccca gatgcagagc 1200
tgcacatctt ccgggtcttt accaacaatc aggtgtctta cacatcttgg tttttggacg 1260
ctttcaacta tgccatccta aagaagattg atgttctaaa ccttagcatc ggcgggcctg 1320
acttcatgga tcatcccttt gttgacaagg tgtgggaatt aacagctaac aatgtaatca 1380
tggtttctgc tatcggcaat gatggacctc tttatggcac tctgaataacccagctgatc 1440
agatggatgt gattggagtg ggtggcattg actttgaaga taacatcgcc cgcttttctt 1500
ccaggggaat gactacctgg gaactaccag gaggctatgg tcgcgtgaaa cctgacattg 1560
tcacctatgg tgccggagtg cggggttccg gtgtgaaagg gggctgccgg gcactctcag 1620
ggaccagtgt cgcttcccca gtggttgctg gggctgtcac cttgttagta agcacagtgc 1680
agaagcggga gctagtgaat cctgccagtg tgaagcaagc cctgattgca tcagcccgga 1740
ggcttcctgg tgttaacatg ttcgagcaag gccatggcaa gctggatctg ctgcgagcct 1800
atcagatcct cagcagctac aaaccacagg cgagcttgag tcctagctac atcgacctga 1860
ctgagtgtcc ctacatgtgg ccttactgtt ctcagcccat ctactatgga ggaatgccaa 1920
caattgttaa tgtcaccatc ctcaatggca tgggagtcac aggaagaatt gtggataagc 1980
ctgagtggcg gccctattta ccacagaatg gagacaacat tgaagtggcc ttctcctact 2040
cctcagtgtt atggccttgg tcaggctacc tggccatctc catttctgtg accaagaagg 2100
cagcttcctg ggaaggcatt gcacagggtc acatcatgat cacggtggct tccccagcag 2160
agacggaagc aaaaaatggt gccgagcata cttccacagt gaagcttccc attaaggtga 2220
agatcattcc cacccctcct cggagcaaga gagtcctctg ggaccagtat cacaacctcc 2280
gctacccccc aggctacttt cccagggaca acttgcggat gaagaatgat cctttagact 2340
ggaatggcga ccatgtccac accaatttca gggacatgta ccagcacctg cgcagcatgg 2400
gctacttcgt ggaggtgctc ggtgccccat tcacgtgctt tgatgctaca cagtatggca 2460
ctttgctcat ggtggatagt gaagaagagt acttcccaga ggagattgcc aagctgagga 2520
gggacgtgga caatggcctt tccctcgtca tcttcagtga ctggtacaac acttctgtta 2580
tgagaaaagt gaagttttac gatgaaaaca caaggcagtg gtggatgcca gatactggag 2640
gagccaacat cccagctctg aacgagctgc tgtctgtgtg gaacatgggg ttcagcgatg 2700
gcctttatga aggggagttt gccctggcga atcatgacat gtattatgca tcgggatgca 2760
gcatcgccaa gtttccagaa gatggtgttg tgatcacaca gactttcaag gaccaaggat 2820
tggaggtctt aaaacaagag acagcagttg ttgaaaatgt tcccattttg gggctttatc 2880
agattccagc tgaaggtggg ggccggatcg tgttgtatgg agattccaat tgcttggatg 2940
acagtcacag acagaaggat tgcttttggc ttctggatgc actccttcag tacacatcat 3000
atggcgtgaa ccctcccagc ctcagccatt cagggaaccg gcagcgccca cccagtggag 3060
ctggcttggc ccctcctgaa aggatggaag gaaaccacct tcatcgatac tccaaggttc 3120
ttgaggccca tctgggagac ccaaaacctc ggcctcttcc agcctgtcca cacttgtcat 3180
gggccaagcc acagcctttg aatgagactg cgcccagtaa tctttggaaa catcagaagc 3240
tgctctccat tgacctggac aaagtagtgt tacccaactt tcgatcgaat cgccctcaag 3300
tgagaccttt gtcccctgga gaaagtggtg cctgggacat tcctggaggg atcatgcctg 3360
gccgctacaa ccaagaggtg ggccagacca tccctgtctt tgccttcctc ggagccatgg 3420
tggccctggc cttctttgtg gtacagatca gcaaggccaa aagccggccg aagcggagga 3480
ggcccagggc aaagcgtcca cagcttacac agcagaccca cccaccaagg accccgtcag 3540
tgtgatcatc acagtggcca gccacagaag ccaacaagcc ttggaccact ctgatggcca 3600
cacagggcat cagaagagca tcctgggagg tgcctatttc caagggaccc catctccagc 3660
ttgtggctgg gttagtgtgt tctccccagg catctctgag ttacatcctg aagtacctca 3720
ctgtgctggg ctcttgacag gaggtgctca gtagctcagc ctccagtggt gtcagcaggc 3780
ccagtgacag tgcaccaaag acacagagcc tggaagggct gtcgggacac actttctaca 3840
taaagcttac aatcctgacc aagcgaagaa atgcttgtta caggctattt tctatattta 3900
ttgtggggag agtcacttta aagacttgta ctgtttggaa gcaaagctgt tgtgtttgtc 3960
agttgagtgc agttttctgc agtgacatca taaggagtca gatcccatga cctttttgat 4020
gagaggacag actgaactga agggcatgtg cacagatctg ggaaatgcaa gccttcgctt 4080
tatttttata agtatcaact gccatcatgt tttgtaattt ggggtcttga tttcaccatt 4140
gttggtgaaa gaaattttca ataaatatgc ataaccttaa aaaaaaaaaa aaaaaaaa 4198
<210>313
<211>409
<212>PRT
<213>Hypocrea lixii
<220>
<223>Alkaline proteinase precursor (ALP) S08.066
<400>313
Met Thr Ser Ile Arg Arg Leu Ala Leu Tyr Leu Gly Ala Leu Leu Pro
1 5 10 15
Ala Val Leu Ala Ala Pro Ala Ala Leu His Lys Lys Pro Glu Ala Val
20 25 30
Pro Asn Lys Phe Ile Val Thr Leu Lys Glu Gly Ala Ser Ile Asp Thr
35 40 45
Asp Ser His Leu Ala Trp Val Thr Asp Ile His Thr Arg Ser Leu Thr
50 55 60
Lys Arg Ser Thr Ala Gly Val Glu Lys Thr Tyr Asn Ile His Thr Trp
65 70 75 80
Asn Ala Tyr Ala Gly Glu Phe Asp Glu Glu Thr Ile Glu Gln Ile Lys
85 90 95
Ser Asn Pro Asp Val Ala Ser Val Glu Pro Asp Tyr Ile Met His Leu
100 105 110
Ser Asp Ile Val Glu Asp Lys Arg Ala Leu Thr Thr Gln Ser Gly Ala
115 120 125
Pro Trp Gly Leu Gly Thr Val Ser His Arg Thr Ser Gly Ser Thr Ser
130 135 140
Tyr Ile Tyr Asp Ser Ser Ala Gly Ala Gly Thr Phe Ala Tyr Val Val
145 150 155 160
Asp Ser Gly Ile Asn Thr Ser His Gln Gln Phe Gly Gly Arg Ala Ser
165 170 175
Leu Gly Tyr Asn Ala Ala Gly Gly Gln His Val Asp Thr Leu Gly His
180 185 190
Gly Thr His Val Ser Gly Thr Ile Gly Gly Ser Thr Tyr Gly Val Ala
195 200 205
Lys Gln Ala Ser Leu Ile Ser Val Lys Val Phe Ala Gly Glu Ser Ala
210 215 220
Ser Thr Ser Val Ile Leu Asp Gly Tyr Asn Trp Ala Val Asn Asp Ile
225 230 235 240
Val Ser Lys Ser Arg Ala Ser Lys Ser Ala Ile Asn Met Ser Leu Gly
245 250 255
Gly Pro Ala Ser Ser Thr Trp Thr Thr Ala Ile Asn Ala Ala Phe Asn
260 265 270
Gln Gly Val Leu Thr Ile Val Ala Ala Gly Asn Gly Asp Ser Leu Gly
275 280 285
Asn Pro Gln Pro Val Ser Gly Thr Ser Pro Ala Asn Val Pro Asn Ala
290 295 300
Ile Thr ValAla Ala Leu Asp Ile Asn Trp Arg Thr Ala Ser Phe Thr
305 310 315 320
Asn Tyr Gly Ala Gly Val Asp Val Phe Ala Pro Gly Val Asn Ile Leu
325 330 335
Ser Ser Trp Ile Gly Ser Asn Thr Ala Thr Asn Thr Ile Ser Gly Thr
340 345 350
Ser Met Ala Thr Pro His Val Val Gly Leu Ala Leu Tyr Leu Gln Ser
355 360 365
Leu Glu Gly Leu Thr Ser Pro Thr Ala Val Thr Asn Arg Ile Lys Ala
370 375 380
Leu Ala Thr Ser Gly Arg ValThr Gly Ser Leu Asn Gly Ser Pro Asn
385 390 395 400
ValIle Ile Phe Asn Gly Asn Ser Ser
405
<210>314
<211>1458
<212>DNA
<213>Hypocrea lixii
<220>
<223>Alkaline proteinase precursor (ALP) S08.066
<400>314
ctcctccact atacaaagca catcacagcc atgaccagca ttcgtcgtct cgctctctat 60
cttggagctt tgctcccggc tgtcctcgct gctcctgcag ctcttcacaa gaagcctgaa 120
gccgtaccca acaagttcat cgtcactctt aaagagggtg cttctattga taccgactct 180
catctcgcct gggtaacgga catccacacc cgttcgttga ccaagcgtag cactgctggt 240
gttgaaaaga cttataacat tcatacttgg aatgcttatg cgggcgagtt tgacgaggag 300
acaattgagc agatcaagtc caaccccgat gttgcgtctg ttgagccaga ctacatcatg 360
catctgtcgg acattgttga agacaagcgt gctctgacca cacaatctgg tgctccttgg 420
ggtctcggca ctgtgtccca tcgcacatct ggatccacaa gctacattta tgatagctca 480
gccggcgccg gcacctttgc ctatgtggtt gactctggta tcaacacttc tcatcagcaa 540
ttcggcgggc gtgccagcct tggctacaat gctgcaggag gacagcatgt cgatactctt 600
ggccatggta ctcatgtttc cggaacaatt ggtggatcta catacggtgt tgctaagcag 660
gcgagcctaa tctctgtcaa ggtctttgct ggtgaaagtg cttccacctc tgttatcctt 720
gacggctata actgggctgt aaacgacatc gtctcgaaaa gccgtgctag caagtctgct 780
attaacatgt cgcttggagg acctgcctca tctacctgga cgaccgccat taacgcagcc 840
tttaaccagg gcgtgcttac cattgtcgcc gctggtaatg gcgactctct gggaaacccc 900
cagccagtct ccggcacttc tcctgctaac gttcctaacg ccataaccgt tgccgcattg 960
gacatcaact ggcgcactgc ttccttcacc aactatggtg ctggcgttga tgtttttgcc 1020
cctggcgtca acattctgtc gtcatggatt ggatccaaca ctgctaccaa cacaatcagc 1080
ggtacttcca tggctacacc tcacgttgtt ggcctggccc tctatcttca atctctcgag 1140
ggtctcacct ctccgaccgc tgtcactaac cgaattaaag ctctggctac atctggtcgt 1200
gtaaccggaa gcctcaacgg cagccccaac gttatcatct tcaacggaaa cagttcttaa 1260
atcgttgatg gagcgaagga tgaagtgaaa tgttcatgga agggcagcac agctgttgct 1320
tggcttgctt agctcattta agaagctaat caccatgatg gagtgcgatg ggtatataag 1380
cttctacagc gttttcaact tagctagatg accctctata gatcgctaaa acaaaaagat 1440
atgacagata cctgtgct 1458
<210>315
<211>1045
<212>PRT
<213>Serratia marcescens
<220>
<223>Extracellular serine protease precursor S08.094
<400>315
Met Ile Leu Asn Lys Arg Leu Lys Leu Ala Tyr Cys Val Phe Leu Gly
1 5 10 15
Cys Tyr Gly Leu Ser Ile His Ser Ser Leu Ala Ala Tyr Gln Asp Pro
20 25 30
Gly Arg Leu Gly Ala Pro Asp Ser Trp Lys Thr Ala Glu Phe Asn Arg
35 40 45
Gln Trp Gly Leu Glu Ala Ile Ser Ala Glu Phe Ala Tyr Ala Arg Gly
50 55 60
Tyr Thr Gly Lys Gly Ile Thr Ile Gly Val Ile Asp Asn Ala Ile Leu
65 70 75 80
Ser His Ser Glu Phe Ser Gly Lys Leu Thr Arg Leu Asp Asn Gly Ser
85 90 95
Tyr Asn Phe Ser Tyr Asp Lys Gln Asp Asn Met Ser Phe Gly Asp His
100 105 110
Gly Thr His Val Ala Gly Ile Ala Ala Ala Lys Arg Asp Gly Ala Gly
115 120 125
Met His Gly Val Ala Phe Asp Ala Asp Ile Ile Gly Thr Lys Leu Asn
130 135 140
Asp Tyr Gly Asn Arg Asn Gly Arg Glu Glu Leu Ile Gln Ser Ala Ala
145 150 155 160
Arg Val Ile Asn Asn Ser Trp Gly Ile Ala Pro Asp Ile Arg Arg Asp
165 170 175
Ala Lys Gly Asp Ile Ile Trp Leu Pro Asn Gly Arg Pro Asp Tyr Val
180 185 190
Ala Phe Val Lys Ser Glu Val Ile Ala Glu Met Met Arg Ser Lys Ser
195 200 205
Ser Val Glu Trp Gly Ser Glu Gln Pro Val Pro Thr Gly Gly His Ser
210 215 220
Ala Met Ser Thr Leu Leu Arg Ala Ala Arg His Gly Lys Leu Ile Val
225 230 235 240
Phe Ser Ala Gly Asn Tyr Asn Asn Tyr Asn Ile Pro Glu Ala Gln Lys
245 250 255
Ser Leu Pro Tyr Ala Phe Pro Asp Val Leu Asn Asn Tyr Leu Ile Val
260 265 270
Thr Asn Leu Ser Asp Glu Asn Gln Leu Ser Val Ser Ser Thr Ser Cys
275 280 285
Gly Gln Thr Ala Ser Tyr Cys Val Ser Ala Pro Gly Ser Asp Ile Tyr
290 295 300
Ser Thr Val Gly Arg Leu Glu Ser Asn Thr Gly Gly Ala Val Asn Arg
305 310 315 320
Glu Ala Tyr Asn Lys Gly Glu Leu Ser Leu Asn Pro Gly Tyr Gly Asn
325 330 335
Lys Ser Gly Thr Ser Met Ala Ala Pro His Val Thr Gly Val Ala Ala
340 345 350
Val Leu Met Gln Arg Phe Pro Tyr Met Ser Ala Asp Gln Ile Ser Ala
355 360 365
Val Ile Lys Thr Thr Ala Thr Asp Leu Gly Val Ala Gly Ile Asp Asn
370 375 380
Leu Phe Gly Trp Gly Arg Val Asn Leu Arg Asp Ala Ile Asn Gly Pro
385 390 395 400
Lys Met Phe Ile Thr Lys Glu Asp Ile Pro Gln Glu Tyr Tyr Val Pro
405 410 415
Gly Ser Tyr Ser Glu Lys Gln Phe Val Val Asn Ile Pro Gly Leu Gly
420 425 430
Asn Ile Val Glu Pro Gly Thr Pro Val Glu Arg Arg Cys Thr Ser Ser
435 440 445
Glu Cys Ser Phe Asp Ser Trp Ser Asn Asp Ile Ser Gly His Gly Gly
450 455 460
Leu Thr Lys Thr Gly Ala Gly Thr Leu Ala Leu Leu Gly Asn Asn Thr
465 470 475 480
Tyr Arg Gly Asp Thr Trp Val Lys Gln Gly Val Leu Ala Ile Asp Gly
485 490 495
Ser Val Ala Ser Asn Val Tyr Ile Glu Asn Ser Gly Thr Leu Ser Gly
500 505 510
Glu Gly Thr Val Gly Ala Phe Arg Ala Ala Arg Ser Gly Ser Val Ala
515 520 525
Pro Gly Asn Gly Ile Gly Thr Leu His Val Leu His Asp Ala Ile Phe
530 535 540
Asp Arg Gly Ser Gln Tyr Asn Val Glu Val Ala Asp Asn Gly Arg Ser
545 550 555 560
Asp Lys Ile Ala Ala Arg Arg Ala Phe Leu Asn Gly Gly Ser Val Asn
565 570 575
Val Ser Leu Glu Arg Ser Gln Asn Leu Leu Ser Gln Asn Glu Ala Gln
580 585 590
Ser Leu Leu Gly Asn Lys Tyr Thr Ile Leu Thr Thr Thr Asp Gly Val
595 600 605
Thr Gly Arg Phe Glu Asn Ala Asn Pro Ser Tyr Pro Phe Val Lys Val
610 615 620
Ala Leu Asp Tyr Arg Gly Asn Asp Val Gly Leu Gly Ile Thr Arg Thr
625 630 635 640
Asp Ala Ser Phe Asp Ser Leu Ala Ser Thr Glu Asn Glu Lys Ala Val
645 650 655
Ala Arg Ala Val Glu Thr Leu Asn Ala Thr Glu Pro Val Thr Glu Thr
660 665 670
Ala Lys Arg Ser Val Ala Ile Pro Ala Ala Glu Glu Ala Asn Leu Leu
675 680 685
Gln Ser Asp Gly Gly Glu Ala Gln Ala Val Asn Glu Glu Ala Ser Ile
690 695 700
Val Ala Gly His Pro Ile Tyr Glu Ser Phe Leu Gly Phe Thr Ser Ala
705 710 715 720
Arg Glu Leu Gln Gln Ala Thr Arg Gln Leu Ser Gly Gln Ile His Ala
725 730 735
Asp Met Ala Ser Ala Gln Ile Asn Glu Ser Arg Tyr Leu Arg Asp Thr
740 745 750
Ala Thr Glu Arg Leu Arg Gln Ala Glu Gly Arg Arg Thr Ala Thr Asp
755 760 765
Ile Lys Ala Asp Asp Asn Gly Ala Trp Ala Lys Leu Leu Gly Ser Trp
770 775 780
Gly His Ala Ser Gly Asn Asp Asn Ala Thr Gly Tyr Gln Thr Ser Thr
785 790 795 800
Tyr Gly Val Leu Leu Gly Leu Asp Ser Glu Leu Phe Gly Asp Gly Arg
805 810 815
Leu Gly Met Met Thr Gly Tyr Thr Arg Thr Ser Leu Asp Gly Gly Tyr
820 825 830
Gln Ser Asp Ala His Ser Asp Asn Tyr His Leu Gly Leu Tyr Gly Asp
835 840 845
Lys Arg Phe Gly Ala Leu Ala Leu Arg Ala Gly Gly Thr Tyr Thr Trp
850 855 860
His Arg Ile Asp Thr Ser Arg Ser Val Asn Tyr Gly Ala Gln Ser Asp
865 870 875 880
Arg Glu Lys Ala Lys Tyr Asn Ala Arg Thr Gly Gln Leu Phe Ile Glu
885 890 895
Ser Gly Tyr Asp Trp Thr Ser Asp Ala Val Asn Leu Glu Pro Phe Ala
900 905 910
Asn Leu Ala Tyr Thr His Tyr Arg Asn Glu Glu Ile Asn Glu Gln Gly
915 920 925
Gly Ala Ala Ala Leu Arg Gly Asp Lys Gln Ser Gln Ser Ala Thr Ala
930 935 940
Ser Thr Leu Gly Leu Arg Ala Asp Thr Glu Trp Gln Thr Asp Ser Val
945 950 955 960
Ala Ile Ala Leu Arg Gly Glu Leu Gly Trp Gln His Gln Tyr Gly Lys
965 970 975
Leu Glu Arg Lys Thr Gln Leu Met Phe Lys Arg Thr Asp Ala Ala Phe
980 985 990
Asp Val Asn Ser Val Pro Val Ser Arg Asp Gly Ala Ile Leu Lys Ala
995 1000 1005
Gly Val Asp Val Ser Ile Asn Lys Asn Ala Val Leu Ser Leu Gly Tyr
1010 1015 1020
Gly Gly Gln Leu Ser Ser Asn His Gln Asp Asn Ser Val Asn Ala Gly
1025 1030 1035 1040
Leu Thr Trp Arg Phe
1045
<210>316
<211>3675
<212>DNA
<213>Serratia marcescens
<220>
<223>Extracellular serine protease precursor S08.094
<400>316
gatccttacg cccggtcccg atggtcggtc attccggcgc aacgggcgaa acgtacattc 60
gcctcctctc cagacgtcct gcgccatagc cgcggcgcga tggcatccct ttgtctgtcc 120
gcgcccattt gcatctctca gcatgtctgt tcgtgcggcc cgcgccgggt ccgcatgtgt 180
tatccgtcgt ttcacgatca taggaggttt cagcgcggtt aaaacgtaag ttttcttctg 240
ggtataaata aacatggagc gtaaaatgat acttaataaa agattgaagt tagcgtattg 300
cgtttttctg ggttgttatg gcttatccat tcattcttct cttgccgctt atcaggatcc 360
cggtcgattg ggcgcgcctg acagttggaa aaccgcggag tttaatcgcc aatgggggct 420
tgaagctatt tccgccgaat tcgcctatgc cagaggctat accggaaaag gtataaccat 480
cggcgttatc gataacgcta ttctttccca ttctgaattc tccggtaaac tgacgcgcct 540
ggataacggc agttataatt tctcgtatga taaacaagat aatatgtctt tcggcgatca 600
cggcacgcac gtagccggta tcgctgcggc taaaagagat ggcgcaggca tgcacggcgt 660
cgctttcgac gcggatatta tcggcaccaa attgaatgat tacggtaatc gcaacggccg 720
tgaagagctg attcagagcg cggctcgcgt catcaataac agctggggga tcgcgccgga 780
tatccggcga gacgccaaag gcgatatcat ctggttgccg aacggcaggc cggactacgt 840
ggcattcgta aaaagcgaag tgatcgccga gatgatgcgc agcaaatcca gcgtggaatg 900
gggcagcgaa caaccggtgc ccaccggcgg gcacagcgcc atgtcgacgc tgctgcgggc 960
ggccaggcac ggcaagctga tcgtcttctc ggcgggcaat tacaacaatt acaatattcc 1020
ggaagcgcaa aagtcactgc cttacgcttt cccggacgta ctgaataatt acctgattgt 1080
gaccaacctg agtgacgaaa atcagttaag cgtttcctcg accagctgcg ggcaaacggc 1140
cagctattgt gtttccgcgc cgggttctga tatttacagc accgtcggcc ggctggagtc 1200
caataccggc ggcgccgtga atcgcgaagc ttataataag ggcgagctgt cgcttaatcc 1260
tggttacggc aacaaatccg gcacctcaat ggcggcgccg catgtgaccg gcgtcgccgc 1320
cgtattgatg cagcgcttcc catatatgag cgccgatcaa atatctgcag tcattaaaac 1380
caccgccacc gatttaggcg ttgccggcat tgataattta ttcggctggg ggcgcgtcaa 1440
tttacgcgat gcgatcaacg gcccgaaaat gtttattacc aaagaggata tcccgcagga 1500
atattatgtg ccgggttcct acagtgagaa acagtttgtg gtgaatatcc ccggcctggg 1560
aaatatcgtc gaacccggca cgccggttga gcggcgctgc acgtcgagcg aatgcagttt 1620
cgattcgtgg agtaacgaca tcagcgggca cggcggcttg accaaaacgg gcgccggtac 1680
gctggcgctg ttgggtaata acacctatcg cggtgatacc tgggtgaaac agggcgtgtt 1740
ggcgatcgac ggttcggtgg catccaacgt ctatatcgaa aatagcggca cattgtccgg 1800
cgagggcacc gtaggcgcct tcagagcggc tcggagcggc agcgtcgcgc cgggcaacgg 1860
catcggcacg ctgcatgtgt tgcacgatgc catttttgat cgcggttcgc agtataacgt 1920
ggaagtggcg gacaatggcc gcagcgataa gatcgccgcg cggcgcgctt tcctcaacgg 1980
cggcagtgtg aatgtcagcc tggaacgcag ccaaaacctg ctgtcgcaga atgaggcgca 2040
gagtctgctg ggcaacaagt acaccatcct gaccacgacg gacggcgtta ccggtagatt 2100
cgaaaacgcc aacccatcgt atccgtttgt taaagtcgcg ctggattatc ggggcaatga 2160
cgtcggcctt ggcatcacgc gcaccgacgc cagctttgac agcctggcca gcactgagaa 2220
cgagaaagcg gtagctcgtg cggtggagac gctcaacgcg acggaaccgg tcacggaaac 2280
ggccaaacgc agcgtggcga tcccggcggc cgaagaggcc aacctgctgc aaagcgatgg 2340
gggcgaggcg caagccgtga acgaagaggc gagcatcgtg gcgggccatc cgatctatga 2400
aagcttcctc ggcttcacct cggccagaga attgcaacag gcgacccgtc aactgtccgg 2460
ccagatccac gcggatatgg cttccgccca gatcaacgaa agccgttacc tgcgcgatac 2520
cgccaccgag cgcttgcgcc aggcggaagg ccgccgcacc gctaccgaca ttaaagcgga 2580
tgacaacggc gcctgggcga aactgctggg tagctggggg catgcttccg gcaacgacaa 2640
cgccaccggt taccagacct ccacctatgg cgtgctgtta ggtctggaca gcgaactgtt 2700
tggcgacggc cggcttggca tgatgaccgg gtatacccgc acttcgctgg atggaggtta 2760
tcagtcagat gctcacagcg acaactacca tctggggctg tacggcgaca aacgcttcgg 2820
cgcgttggcg ctgcgagcgg gcggcaccta tacctggcat cgcatcgaca cctcgcgttc 2880
ggtgaactac ggcgcgcagt cggatcgcga gaaggccaag tataacgcgc gcaccggtca 2940
gctgttcatc gaaagcggct acgattggac gagcgatgcg gtcaaccttg agccgttcgc 3000
caacctggcg tatacccatt accgtaacga ggagatcaac gagcaaggcg gggcagcggc 3060
gctgcgcggc gacaaacaaa gtcagtccgc caccgcctcg acgttgggtc tgcgcgccga 3120
caccgagtgg caaaccgaca gcgtggcgat cgcgctgcgc ggcgagctgg gttggcagca 3180
tcagtacggc aagctggagc gtaaaacgca gctgatgttc aaacgcactg atgcggcgtt 3240
cgacgtgaac agcgtgcctg tttctcgcga tggcgcgatt ctgaaagcgg gcgtcgatgt 3300
atcgattaac aaaaacgccg tcctgtccct tggctacggc gggcagctgt cgtccaacca 3360
ccaggacaac agcgtcaacg ccggtctgac ctggcgcttc tgatcgcggt tgcatgcttt 3420
gctttattca gccctcatcg agacgatggg ggcttttttt tcgcggccag ccggccttct 3480
gcggcatgag cgatactttc tccgcgacct attcccccgg ccggtgttcg ccgccgggtt 3540
aatcaatcag aggaaaacaa acgctgctgc caatagcgca attgcgtcag cgtgatcggc 3600
gcagcggcgg caatgaccgc gcgattttcc aacgcaatct cccgcgcacg ttgcaacggc 3660
tcggcgctga ggcct 3675
<210>317
<211>1441
<212>PRT
<213>Drosophila melanogaster
<220>
<223>Tripeptidyl-peptidase II(TPP-II)
S08.090
<400>317
Met Phe Asn Arg Phe Arg Leu Val His Lys Gln Leu Arg Leu Tyr Lys
1 5 10 15
Asn Phe Gly Leu Leu Gly Gln Lys Ala Ser Val Gly Leu Thr Leu Pro
20 25 30
Ile Ile Ser Leu Ser Arg Pro Tyr Met Ala Tyr Met Gly Thr Glu Arg
35 40 45
Ser Val Val Met Ile Thr Ala Pro Ala Thr Lys Glu Phe Ala Glu Ser
50 55 60
Ser Glu Arg Ser Asn Ser Ser Lys Lys Thr Thr Asn Lys Glu Gln Ser
65 70 75 80
Asp Lys Ser Ala Glu Ser Arg Met Ala Thr Ser Gly Ile Val Glu Ser
85 90 95
Phe Pro Thr Gly Ala Leu Val Pro Lys Ala Glu Thr Gly Val Leu Asn
100 105 110
Phe Leu Gln Lys Tyr Pro Glu Tyr Asp Gly Arg Asp ValThr Ile Ala
115 120 125
Ile Phe Asp Ser Gly Val Asp Pro Arg Ala Thr Gly Leu Glu Thr Leu
130 135 140
Cys Asp Gly Lys Thr Val Lys Val Ile Glu Arg Tyr Asp Cys Ser Gly
145 150 155 160
Cys Gly Asp Val Asp Met Lys Lys Lys Val Thr Pro Asp Glu Asn Gly
165 170 175
Asn Ile Lys Gly Leu Ser Gly Asn Ser Leu Lys Leu Ser Pro Glu Leu
180 185 190
Met Ala Leu Asn Thr Asp Pro Glu Lys Ala Val Arg Val Gly Leu Lys
195 200 205
Ser Phe Ser Asp Leu Leu Pro Ser Lys Val Arg Asn Asn Ile Val Ala
210 215 220
Gln Ala Lys Leu Lys His Trp Asp Lys Pro His Lys Thr Ala Thr Ala
225 230 235 240
Asn Ala Ser Arg Lys Ile Val Glu Phe Glu Ser Gln Asn Pro Gly Glu
245 250 255
Ala Ser Lys Leu Pro Trp Asp Lys Lys Ile Leu Lys Glu Asn Leu Asp
260 265 270
Phe Glu Leu Glu Met Leu Asn Ser Tyr Glu Lys Val Tyr Gly Asp Ile
275 280 285
Lys Thr Ser Tyr Asp Cys Ile Leu Phe Pro Thr Ala Asp Gly Trp Leu
290 295 300
Thr Ile Val Asp Thr Thr Glu Gln Gly Asp Leu Asp Gln Ala Leu Arg
305 310 315 320
Ile Gly Glu Tyr Ser Arg Thr His Glu Thr Arg Asn Val Asp Asp Phe
325 330 335
Leu Ser Ile Ser Val Asn Val His Asp Glu Gly Asn Val Leu Glu Val
340 345 350
Val Gly Met Ser Ser Pro His Gly Thr His Val Ser Ser Ile Ala Ser
355 360 365
Gly Asn His Ser Ser Arg Asp Val Asp Gly Val Ala Pro Asn Ala Lys
370 375 380
Ile Val Ser Met Thr Ile Gly Asp Gly Arg Leu Gly Ser Met Glu Thr
385 390 395 400
Gly Thr Ala Leu Val Arg Ala Met Thr Lys Val Met Glu Leu Cys Arg
405 410 415
Asp Gly Arg Arg Ile Asp Val Ile Asn Met Ser Tyr Gly Glu His Ala
420 425 430
Asn Trp Ser Asn Ser Gly Arg Ile Gly Glu Leu Met Asn Glu Val Val
435 440 445
Asn Lys Tyr Gly Val Val Trp Val Ala Ser Ala Gly Asn His Gly Pro
450 455 460
Ala Leu Cys Thr Val Gly Thr Pro Pro Asp Ile Ser Gln Pro Ser Leu
465 470 475 480
Ile Gly Val Gly Ala Tyr Val Ser Pro Gln Met Met Glu Ala Glu Tyr
485 490 495
Ala Met Arg Glu Lys Leu Pro Gly Asn Val Tyr Thr Trp Thr Ser Arg
500 505 510
Asp Pro Cys Ile Asp Gly Gly Gln Gly Val Thr Val Cys Ala Pro Gly
515 520 525
Gly Ala Ile Ala Ser Val Pro Gln Phe Thr Met Ser Lys Ser Gln Leu
530 535 540
Met Asn Gly Thr Ser Met Ala Ala Pro His Val Ala Gly Ala Val Ala
545 550 555 560
Leu Leu Ile Ser Gly Leu Lys Gln Gln Asn Ile Glu Tyr Ser Pro Tyr
565 570 575
Ser Ile Lys Arg Ala Ile Ser Val Thr Ala Thr Lys Leu Gly Tyr Val
580 585 590
Asp Pro Phe Ala Gln Gly His Gly Leu Leu Asn Val Glu Lys Ala Phe
595 600 605
Glu His Leu Thr Glu His Arg Gln Ser Lys Asp Asn Met Leu Arg Phe
610 615 620
Ser Val Arg Val Gly Asn Asn Ala Asp Lys Gly Ile His Leu Arg Gln
625 630 635 640
Gly Val Gln Arg Asn Ser Ile Asp Tyr Asn Val Tyr Ile Glu Pro Ile
645 650 655
Phe Tyr Asn Asp Lys Glu Ala Asp Pro Lys Asp Lys Phe Asn Phe Asn
660 665 670
Val Arg Leu Asn Leu Ile Ala Ser Gln Pro Trp Val Gln Cys Gly Ala
675 680 685
Phe Leu Asp Leu Ser Tyr Gly Thr Arg Ser Ile Ala Val Arg Val Asp
690 695 700
Pro Thr Gly Leu Gln Pro Gly Val His Ser Ala Val Ile Arg Ala Tyr
705 710 715 720
Asp Thr Asp Cys Val Gln Lys Gly Ser Leu Phe Glu Ile Pro Val Thr
725 730 735
Val Val Gln Pro His Val Leu Glu Ser Asp Gln Asn Thr Pro Val Phe
740 745 750
Glu Pro Ala Ser Ser Lys Gly Asp Asn Ser Val Glu Phe Gln Pro Asn
755 760 765
Thr Ile Gln Arg Asp Phe Ile Leu Val Pro Glu Arg Ala Thr Trp Ala
770 775 780
Glu Leu Arg Met Arg Ile Thr Asp Pro Asn Arg Gly Glu Asp Ile Gly
785 790 795 800
Lys Phe Phe Val His Thr Asn Gln Leu Leu Pro Lys Gln Ser Cys Arg
805 810 815
Lys Leu Glu Thr Met Lys Ile Val Ser Val Gly Ser Glu Asn Glu Ser
820 825 830
Ile Met Ala Phe Lys Val Lys Ser Gly Arg Ile Leu Glu Leu Cys Ile
835 840 845
Ala Lys Tyr Trp Ser Asn Tyr Gly Gln Ser His Leu Lys Tyr Ser Leu
850 855 860
Arg Phe Arg Gly Val Glu Ala His Asn Pro Asn Ala Tyr Val Met His
865 870 875 880
Ala Gly Arg Gly Ile His Lys Leu Glu Ile Glu Ala Leu Val Ala Glu
885 890 895
Asp Val Gln Pro Gln Leu Gln Leu Lys Asn Ala Glu Val Val Leu Lys
900 905 910
Pro Thr Glu Ala Lys Ile Ser Pro Leu Ser Ala Thr Arg Asp Val Ile
915 920 925
Pro Asp Gly Arg Gln Val Tyr Gln Asn Leu Leu Ala Phe Asn Leu Asn
930 935 940
Val Ala Lys Ala Ala Asp Val Ser Ile Tyr Ala Pro Ile Phe Asn Asp
945 950 955 960
Leu Leu Tyr Glu Ala Glu Phe Glu Ser Gln Met Trp Met Leu Phe Asp
965 970 975
Ala Asn Lys Ala Leu Val Ala Thr Gly Asp Ala His Ser His Thr Ser
980 985 990
Phe Thr Lys Leu Asp Lys Gly Glu Tyr Thr Ile Arg Leu Gln Val Arg
995 1000 1005
His Glu Lys Arg Asp Leu Leu Glu Lys Ile Ser Glu Ala Asn Leu Val
1010 1015 1020
Ala Ser Phe Lys Leu Thr Ser Pro Leu Thr Leu Asp Phe Tyr Glu Asn
1025 1030 1035 1040
Tyr Asn Gln Cys Ile Val Gly Gly Arg Lys Tyr Val Ser Ser Pro Leu
1045 1050 1055
Arg Leu Ser Thr Arg Val Leu Tyr Ile Ala Pro Ile Thr Gln Glu Arg
1060 1065 1070
Leu Thr Lys Ala Asn Leu Pro Ala Gln Cys Ala Trp Leu Ser Gly Asn
1075 1080 1085
Leu Val Phe Pro Gln Asp Glu Val Gly Arg Arg Val Ala Gln His Pro
1090 1095 1100
Phe Thr Tyr Ile Leu Asn Pro Ala Glu Lys Lys Ser His Thr Asn Gly
1105 1110 1115 1120
Ser Ser Asn Gly Ser Ser Ala Ala Gly Ser Thr Ala Thr Ala Ala Ala
1125 1130 1135
Val Thr Thr Ala Asn Gly Ala Lys Pro Lys Ala Pro Ala Thr Pro Gln
1140 1145 1150
Ala Ala Thr Ser Val Thr Asn Pro Ala Ala Gly Asp Gly Ile Ser Val
1155 1160 1165
Gln Asn Asp Pro Pro Val Asp Ser Ser Gly Ser Pro Ala Ser Pro Lys
1170 1175 1180
Lys Gly Lys Ala Asn Ala Asp Asp Tyr Ala Glu Ser Phe Arg Asp Phe
1185 1190 1195 1200
Gln Cys Ser Gln Ile Val Lys Cys Glu Leu Glu Met Ala Glu Lys Ile
1205 1210 1215
Tyr Asn Asp Val Val Ala Ala His Pro Lys His Leu Gln Ala Asn Leu
1220 1225 1230
Leu Leu Ile Gln Asn Ile Glu Ser Asn Gln Leu Lys Ser Gln Leu Pro
1235 1240 1245
Leu Thr Phe Val Asn Ala Gln Lys Thr Ser Pro Pro Glu Ala Gly Glu
1250 1255 1260
Ser Ala Asp Lys Gln Lys Glu Asp Gln Lys Lys Val Arg Ser Ala Leu
1265 1270 1275 1280
Glu Arg Ile Val Lys Leu Ala Asp Lys Val Ile Gln Glu Thr Asp Ser
1285 1290 1295
Glu Ala Leu Leu Ser Tyr Tyr Gly Leu Lys Asn Asp Thr Arg Ala Asp
1300 1305 1310
Ala Ala Lys Ile Lys Thr Asn Met Asp Lys Gln Lys Asn Thr Leu Ile
1315 1320 1325
Glu Ala Leu Ser Lys Lys Gly Ile Ala Val Ala Lys Leu Ala Val Leu
1330 1335 1340
Asp Asp Cys Ile Lys Asp Ser Leu Ala Glu Ile Asn Glu Leu Tyr Thr
1345 1350 1355 1360
Glu Ile Ile Lys Phe Val Asp Ala Asn Asp Ser Lys Ala Ile Gln Phe
1365 1370 1375
Ala Leu Trp His Ala Tyr Ala His Gly His Tyr Gly Arg Met Tyr Lys
1380 1385 1390
Tyr Val Val Lys Leu Ile Glu Glu Lys Arg Thr Arg Asp His Phe Val
1395 1400 1405
Glu Leu Ala Ala Ile Asn Gly Ala Leu Gly His Glu His Ile Arg Thr
1410 1415 1420
Val Ile Asn Arg Met Met Ile Thr Ala Phe Pro Ser Ser Phe Arg Leu
1425 1430 1435 1440
Phe
<210>318
<211>4309
<212>DNA
<213>Drosophila melanogaster
<220>
<223>Tripeptidyl-peptidase II(TPP-II)S08.090
<400>318
ggcccgagta ccaatccgat tcttcattga acagctacga aggaattcgc agagagttct 60
gagcgcagca atagcagcaa gaagactacc aacaaagagc agagcgacaa atcagcggag 120
agcaggatgg ccaccagcgg tattgtcgag tcatttccca cgggcgcact ggtgcccaag 180
gcggagacgg gagtcctgaa ctttctgcag aagtacccgg agtacgatgg acgcgacgtc 240
accatagcca tcttcgattc cggcgtcgat ccccgggcaa cgggactgga gacgctgtgc 300
gatggaaaga ccgttaaagt aatagagcgg tacgactgtt ccggatgcgg cgacgtggac 360
atgaagaaga aggtgacgcc ggacgagaac ggcaacatta agggcctgtc cggaaactcg 420
ctcaagctga gtccggagct gatggctctg aacacagatc cggagaaggc ggtgcgggta 480
ggcctcaaga gcttcagcga tttgttgccc tccaaggtgc ggaacaacat tgtggcccag 540
gccaagctga agcactggga caagccgcac aagacggcca ctgcaaacgc cagtcgcaag 600
attgttgaat ttgagtcaca aaatccagga gaagcctcaa aactgccctg ggacaagaag 660
atattgaaag agaatctaga ctttgagcta gagatgttga atagctacga gaaggtgtac 720
ggcgacatta agacctccta cgattgcatc ctttttccca cggccgacgg atggctgaca 780
atcgtcgaca ccacggagca gggcgatctg gatcaggctc tgcgcattgg cgagtattcc 840
cgcacgcacg agacccgcaa tgtggacgac tttctttcca tatcggtaaa cgtccacgac 900
gagggcaacg tactggaggt ggtcggcatg agctcacccc acggcaccca cgtatcgtct 960
attgccagcg gcaaccatag ctcccgggac gtggatggcg tggcgccgaa tgctaagatc 1020
gtttctatga ccatcggtga tggtcgactt gggtctatgg agaccggcac ggcactagtg 1080
cgtgccatga ctaaggtgat ggaactatgt cgcgatggca gacgcatcga tgtgatcaac 1140
atgagctatg gcgagcatgc caattggtca aattccggcc gcattgggga gctcatgaac 1200
gaggttgtca acaagtatgg cgtggtgtgg gtggcatcgg ccggcaacca tggtccggca 1260
ctttgtactg tgggaactcc gccggacatc agccagccca gtttgatcgg cgtgggcgcg 1320
tacgtatcac cccaaatgat ggaggccgag tatgcgatgc gagaaaagct gcccgggaac 1380
gtgtacacct ggacatcgcg agatccctgc atcgacggag gtcaaggcgt taccgtatgc 1440
gctccgggcg gagccattgc gtccgtgcca cagtttacta tgagcaagtc ccagctgatg 1500
aacggtacca gcatggcggc acctcacgtc gccggcgcag tggcgctgct catctccggt 1560
ctgaagcaac agaacatcga gtattcgccg tacagtatta agcgagcgat cagcgtcact 1620
gccaccaaac tgggctatgt agatcccttt gctcagggcc atggcttgct caatgtcgag 1680
aaggcattcg agcatttgac ggagcaccgc cagtccaagg ataatatgct caggttctcc 1740
gtgcgcgtgg gcaacaacgc agacaagggc attcatttgc gtcagggcgt gcagcgtaac 1800
tccatcgatt acaacgttta tatagagccc atcttctaca atgacaagga ggcggatccc 1860
aaggacaagt ttaacttcaa tgtacggcta aatctgattg cctcgcagcc gtgggtgcag 1920
tgtggagctt tcttggatct cagctatggc acccgctcca ttgccgtgcg cgtcgatccc 1980
actggactcc agccaggcgt tcacagcgct gtgattcggg cttatgacac tgactgcgta 2040
cagaagggtt ctctctttga gattcctgtc acggtggtgc agccccatgt gctggagtcg 2100
gatcagaaca cgcccgtctt cgaacccgcc tcttccaagg gagacaacag cgtggagttt 2160
cagccaaaca ccattcaaag agacttcatc ctggtgccag aacgtgctac ttgggcggag 2220
ttgcgtatgc gtataaccga tcccaatcgt ggcgaggaca ttggaaagtt ctttgtacac 2280
acgaaccaac ttcttcccaa gcaatcctgt cgtaagcttg agaccatgaa gatcgtatcg 2340
gttggctcgg aaaacgaatc tataatggct tttaaagtta agtctggcag gattctggag 2400
ctatgcattg ccaagtactg gtccaactac ggccagagtc acctgaagta cagcttgcgt 2460
ttccgtggcg tggaagcgca caatcccaat gcctacgtca tgcatgcggg caggggaatt 2520
cataagctgg agattgaggc cctggttgcc gaggatgtgc agccccagct acagctaaag 2580
aatgccgaag tggtgctaaa accgaccgag gccaagatct cgccgctaag cgccacacga 2640
gacgtcatcc cagatggacg tcaggtgtat cagaacctgc tggcctttaa cttgaacgtg 2700
gccaaggccg cagatgtgtc gatatacgca ccgatcttta acgacttgtt gtatgaagcg 2760
gagtttgagt cacagatgtg gatgcttttt gatgcaaaca aggccctggt ggccaccggc 2820
gatgctcact cccacacttc cttcacaaag ctcgataagg gcgaatacac aatcagactg 2880
caggtgcgtc acgagaagcg cgacctgctg gagaagatct cggaggcaaa tctggtggcc 2940
tcattcaagc tgactagccc cctcaccctt gatttctatg agaactacaa tcagtgcatc 3000
gtgggaggtc gtaaatacgt ctcgagtccg ctaaggctgt ccactcgggt gctttatatc 3060
gctcctatta cccaagagcg acttaccaag gccaatctgc ccgctcaatg cgcctggctg 3120
agcggcaatc tggtattccc gcaagatgag gtcggacggc gagtggctca gcatccattt 3180
acttacatcc tcaatcccgc tgagaaaaag tcacacacga atggctcgag caacggttcc 3240
agcgctgcag gatccacagc tacggcagct gctgtcacca ctgctaatgg cgcaaaaccg 3300
aaagctccgg cgactccaca agcggccacc tcagtgacca atcctgcggc cggcgatgga 3360
atttctgttc agaacgaccc gcctgtggac agcagtggga gtcccgcctc acccaaaaag 3420
ggcaaggcca atgccgacga ttatgccgaa agttttcgcg actttcaatg ttcgcagatt 3480
gtcaagtgtg aactggaaat ggcagagaaa atctataatg atgtagtcgc tgcccatccc 3540
aagcatttgc aggcaaacct gctacttatc cagaacatcg agtccaatca gctgaagtcg 3600
caactgccgc tgactttcgt caatgctcaa aagacatcgc caccagaggc aggcgagagc 3660
gccgacaagc aaaaggagga tcaaaagaaa gtacgaagcg ccctggagcg aattgttaag 3720
ctggccgaca aggtgatcca ggagaccgat tccgaggcac tgctctccta ctacggtctg 3780
aagaacgaca ctcgtgccga tgcagccaag ataaagacca acatggacaa acagaaaaac 3840
actctcattg aggcgctaag caagaagggc attgctgtgg caaagctggc tgtattagac 3900
gactgtatca aggatagcct ggctgagatc aacgagctgt atacggagat tatcaagttc 3960
gtggacgcca acgactccaa ggccatccag ttcggtctgt ggcacgccta tgcccatggc 4020
cactacggtc gcatgtacaa gtatgtggtt aagctaatcg aggagaagcg aacccgcgat 4080
cacttcgtgg agctggccgc catcaacggc gccctgggtc acgagcacat ccgtactgtc 4140
atcaaccgta tgatgatcac tgcctttccc agcagcttcc gtttgttctg agcactttcc 4200
gacagaatac gcaacaaata ttcacgcttc gaagcgatca ttgttttgta taactaatgt 4260
aatccatatg tatatgaata aatgctatta actgaaaaaa aaaaaaaaa 4309
<210>319
<211>806
<212>PRT
<213>Bacillus subtilis
<220>
<223>Minor extracellular protease vpr precursor
S08.114
<400>319
Met Lys Lys Gly Ile Ile Arg Phe Leu Leu Val Ser Phe Val Leu Phe
1 5 10 15
Phe Ala Leu Ser Thr Gly Ile Thr Gly Val Gln Ala Ala Pro Ala Ser
20 25 30
Ser Lys Thr Ser Ala Asp Leu Glu Lys Ala Glu Val Phe Gly Asp Ile
35 40 45
Asp Met Thr Thr Ser Lys Lys Thr Thr Val Ile Val Glu Leu Lys Glu
50 55 60
Lys Ser Leu Ala Glu Ala Lys Glu Ala Gly Glu Ser Gln Ser Lys Ser
65 70 75 80
Lys Leu Lys Thr Ala Arg Thr Lys Ala Lys Asn Lys Ala Ile Lys Ala
85 90 95
ValLys Asn Gly Lys Val Asn Arg Glu Tyr Glu Gln Val Phe Ser Gly
100 105 110
Phe Ser Met Lys Leu Pro Ala Asn Glu Ile Pro Lys Leu Leu Ala Val
115 120 125
Lys Asp Val Lys Ala Val Tyr Pro Asn Val Thr Tyr Lys Thr Asp Asn
130 135 140
Met Lys Asp Lys Asp Val Thr Ile Ser Glu Asp Ala Val Ser Pro Gln
145 150 155 160
Met Asp Asp Ser Ala Pro Tyr Ile Gly Ala Asn Asp Ala Trp Asp Leu
165 170 175
Gly Tyr Thr Gly Lys Gly Ile Lys Val Ala Ile Ile Asp Thr Gly Val
180 185 190
Glu Tyr Asn His Pro Asp Leu Lys Lys Asn Phe Gly Gln Tyr Lys Gly
195 200 205
Tyr Asp Phe Val Asp Asn Asp Tyr Asp Pro Lys Glu Thr Pro Thr Gly
210 215 220
Asp Pro Arg Gly Glu Ala Thr Asp His Gly Thr His Val Ala Gly Thr
225 230 235 240
Val Ala Ala Asn Gly Thr Ile Lys Gly Val Ala Pro Asp Ala Thr Leu
245 250 255
Leu Ala Tyr Arg Val Leu Gly Pro Gly Gly Ser Gly Thr Thr Glu Asn
260 265 270
Val Ile Ala Gly Val Glu Arg Ala Val Gln Asp Gly Ala Asp Val Met
275 280 285
Asn Leu Ser Leu Gly Asn Ser Leu Asn Asn Pro Asp Trp Ala Thr Ser
290 295 300
Thr Ala Leu Asp Trp Ala Met Ser Glu Gly Val Val Ala Val Thr Ser
305 310 315 320
Asn Gly Asn Ser Gly Pro Asn Gly Trp Thr Val Gly Ser Pro Gly Thr
325 330 335
Ser Arg Glu Ala Ile Ser Val Gly Ala Thr Gln Leu Pro Leu Asn Glu
340 345 350
Tyr Ala Val Thr Phe Gly Ser Tyr Ser Ser Ala Lys Val Met Gly Tyr
355 360 365
Asn Lys Glu Asp Asp Val Lys Ala Leu Asn Asn Lys Glu Val Glu Leu
370 375 380
Val Glu Ala Gly Ile Gly Glu Ala Lys Asp Phe Glu Gly Lys Asp Leu
385 390 395 400
Thr Gly Lys Val Ala Val Val Lys Arg Gly Ser Ile Ala Phe Val Asp
405 410 415
Lys Ala Asp Asn Ala Lys Lys Ala Gly Ala Ile Gly Met Val Val Tyr
420 425 430
Asn Asn Leu Ser Gly Glu Ile Glu Ala Asn Val Pro Gly Met Ser Val
435 440 445
Pro Thr Ile Lys Leu Ser Leu Glu Asp Gly Glu Lys Leu Val Ser Ala
450 455 460
Leu Lys Ala Gly Glu Thr Lys Thr Thr Phe Lys Leu Thr Val Ser Lys
465 470 475 480
Ala Leu Gly Glu Gln ValAla Asp Phe Ser Ser Arg Gly Pro Val Met
485 490 495
Asp Thr Trp Met Ile Lys Pro Asp Ile Ser Ala Pro Gly Val Asn Ile
500 505 510
Val Ser Thr Ile Pro Thr His Asp Pro Asp His Pro Tyr Gly Tyr Gly
515 520 525
Ser Lys Gln Gly Thr Ser Met Ala Ser Pro His Ile Ala Gly Ala Val
530 535 540
Ala Val Ile Lys Gln Ala Lys Pro Lys Trp Ser Val Glu Gln Ile Lys
545 550 555 560
Ala Ala Ile Met Asn Thr Ala Val Thr Leu Lys Asp Ser Asp Gly Glu
565 570 575
Val Tyr Pro His Asn Ala Gln Gly Ala Gly Ser Ala Arg Ile Met Asn
580 585 590
Ala Ile Lys Ala Asp Ser Leu Val Ser Pro Gly Ser Tyr Ser Tyr Gly
595 600 605
Thr Phe Leu Lys Glu Asn Gly Asn Glu Thr Lys Asn Glu Thr Phe Thr
610 615 620
Ile Glu Asn Gln Ser Ser Ile Arg Lys Ser Tyr Thr Leu Glu Tyr Ser
625 630 635 640
Phe Asn Gly Ser Gly Ile Ser Thr Ser Gly Thr Ser Arg Val Val Ile
645 650 655
Pro Ala His Gln Thr Gly Lys Ala Thr Ala Lys Val Lys Val Asn Thr
660 665 670
Lys Lys Thr Lys Ala Gly Thr Tyr Glu Gly Thr Val Ile Val Arg Glu
675 680 685
Gly Gly Lys Thr Val Ala Lys Val Pro Thr Leu Leu Ile Val Lys Glu
690 695 700
Pro Asp Tyr Pro Arg Val Thr Ser Val Ser Val Ser Glu Gly Ser Val
705 710 715 720
Gln Gly Thr Tyr Gln Ile Glu Thr Tyr Leu Pro Ala Gly Ala Glu Glu
725 730 735
Leu Ala Phe Leu Val Tyr Asp Ser Asn Leu Asp Phe Ala Gly Gln Ala
740 745 750
Gly Ile Tyr Lys Asn Gln Asp Lys Gly Tyr Gln Tyr Phe Asp Trp Asp
755 760 765
Gly Thr Ile Asn Gly Gly Thr Lys Leu Pro Ala Gly Glu Tyr Tyr Leu
770 775 780
Leu Ala Tyr Ala Ala Asn Lys Gly Lys Ser Ser Gln Val Leu Thr Glu
785 790 795 800
Glu Pro Phe Thr Val Glu
805
<210>320
<211>2931
<212>DNA
<213>Bacillus subtilis
<220>
<223>Minor extracellular protease vpr precursor S08.114
<400>320
gagatatcca catgtccaca aactctatct atattttgta tacgaagtat attcctaact 60
atatatatac acaggtttat tcacttatac acagggttct gtgtataact ccttcgttat 120
acacaaacaa aatccaataa atggtccaaa tgacacaagg atttttttga attttcaaga 180
aatatatact agatctttca cattttttct aaatacaaag ggggaaacac attgaaaaag 240
gggatcattc gctttctgct tgtaagtttc gtcttatttt ttgcgttatc cacaggcatt 300
acgggcgttc aggcagctcc ggcttcttca aaaacgtcgg ctgatctgga aaaagccgag 360
gtattcggtg atatcgatat gacgacaagc aaaaaaacaa ccgttatagt ggaattaaaa 420
gaaaaatcct tggcagaagc gaaggaagcg ggagaaagcc aatcgaaaag caagctgaaa 480
accgctcgca ccaaagcaaa aaacaaagca atcaaagcag tgaaaaacgg aaaagtaaac 540
cgggaatatg agcaggtatt ctcaggcttc tctatgaagc ttccagctaa tgagattcca 600
aaacttctag cggtaaaaga cgttaaggca gtgtacccga acgtcacata taaaacagac 660
aatatgaagg ataaagacgt cacaatctcc gaagacgccg tatctccgca aatggatgac 720
agtgcgcctt atatcggagc aaacgatgca tgggatttag gctacacagg aaaaggcatc 780
aaggtggcga ttattgacac tggggttgaa tacaatcacc cagatctgaa gaaaaacttt 840
ggacaatata aaggatacga ttttgtggac aatgattacg atccaaaaga aacaccaacc 900
ggcgatccga ggggcgaggc aactgaccat ggcacacacg tagccggaac tgtggctgca 960
aacggaacga ttaaaggcgt agcgcctgat gccacacttc ttgcttatcg tgtgttaggg 1020
cctggcggaa gcggcacaac ggaaaacgtc atcgcgggcg tggaacgtgc agtgcaggac 1080
ggggcagatg tgatgaacct gtctctcgga aactctttaa acaacccgga ctgggcgaca 1140
agcacagcgc ttgactgggc catgtcagaa ggcgttgtcg ctgttacctc aaacggcaac 1200
agcggaccga acggctggac agtcggatcg ccgggcacat caagagaagc gatttctgtc 1260
ggtgcgactc agctgccgct caatgaatac gccgtcactt tcggctccta ctcttcagca 1320
aaagtgatgg gctacaacaa agaggacgac gtcaaagcgc tcaataacaa agaagttgag 1380
cttgtcgaag cgggaatcgg cgaagcaaag gattttgaag ggaaagacct gacaggcaaa 1440
gtcgccgttg tcaaacgagg cagcattgca tttgtggata aagcggataa cgctaaaaaa 1500
gccggtgcaa tcggcatggt tgtgtataac aacctctctg gagaaattga agccaatgtg 1560
ccaggcatgt ctgtcccaac gattaagctt tcattagaag acggcgaaaa actcgtcagc 1620
gccctgaaag ctggtgagac aaaaacaaca ttcaagttga cggtctcaaa agcgctcggt 1680
gaacaagtcg ctgatttctc atcacgcggc cctgttatgg atacgtggat gattaagcct 1740
gatatttccg cgccaggggt caatatcgtg agcacgatcc caacacacga tcctgaccat 1800
ccatacggct acggatcaaa acaaggaaca agcatggcat cgcctcatat tgccggagcg 1860
gttgccgtta ttaaacaagc caaaccaaag tggagcgttg aacagattaa agccgccatc 1920
atgaataccg ctgtcacttt aaaggatagc gatggggaag tatatccgca taacgctcaa 1980
ggcgcaggca gcgcaagaat tatgaacgca atcaaagccg attcgctcgt ctcacctgga 2040
agctattcat acggcacgtt cttgaaggaa aacggaaacg aaacaaaaaa tgaaacgttt 2100
acgattgaaa atcaatcttc cattagaaag tcatacacac ttgaatactc atttaatggc 2160
agcggcattt ccacatccgg cacaagccgt gttgtgattc cggcacatca aaccgggaaa 2220
gccactgcaa aagtaaaggt caatacgaag aaaacaaaag ctggcaccta tgaaggaacg 2280
gttatcgtca gagaaggcgg aaaaacggtc gctaaggtac ctacattgct gattgtgaaa 2340
gagcccgatt atccgagagt cacatctgtc tctgtcagcg aagggtctgt acaaggtacc 2400
tatcaaattg aaacctacct tcctgcggga gcggaagagc tggcgttcct cgtctatgac 2460
agcaaccttg atttcgcagg ccaagccggc atttataaaa accaagataa aggttaccag 2520
tactttgact gggacggcac gattaatggc ggaaccaaac ttccggccgg agagtattac 2580
ttgctcgcat atgccgcgaa caaaggcaag tcaagccagg ttttgaccga agaacctttc 2640
actgttgaat aagaaaaagc cctgccgatt cggcagggct ttttaaagat cagtcagcaa 2700
acgcctcctg caataagcga tacgatcgga gcttatcttc aaaatgatgc gtgatggtca 2760
ccaccatgat ttcctctgtt tcatacgcgt tactcaaagc taacagccgc tccttaacct 2820
gttctttcgt accaacaatc attcgatttc gattatcagc aattcgtctc tgttcataag 2880
gagaatacgt attttccgaa cagcttcata cgaggactcc tctaaggata c 2931
<210>321
<211>1902
<212>PRT
<213>Lactococcus lactis subsp.cremoris
<220>
<223>PIII-type proteinase precursor(Lactocepin)
S08.116
<400>321
Met Gln Arg Lys Lys Lys Gly Leu Ser Ile Leu Leu Ala Gly Thr Val
1 5 10 15
Ala Leu Gly Ala Leu Ala Val Leu Pro Val Gly Glu Ile Gln Ala Lys
20 25 30
Ala Ala Ile Ser Gln Gln Thr Lys Gly Ser Ser Leu Ala Asn Thr Val
35 40 45
Thr Ala Ala Thr Ala Lys Gln Ala Ala Thr Asp Thr Thr Ala Ala Thr
50 55 60
Thr Asn Gln Ala Ile Ala Thr Gln Leu Ala Ala Lys Gly Ile Asp Tyr
65 70 75 80
Asn Lys Leu Asn Lys Val Gln Gln Gln Asp Ile Tyr Val Asp Val Ile
85 90 95
Val Gln Met Ser Ala Ala Pro Ala Ser Glu Asn Gly Ile Leu Arg Thr
100 105 110
Asp Tyr Ser Ser Thr Ala Glu Ile Gln Gln Glu Thr Asn Lys ValIle
115 120 125
Ala Ala Gln Ala Ser Val Lys Ala Ala Val Glu Gln Val Thr Gln Gln
130 135 140
Thr Ala Gly Glu Ser Tyr Gly Tyr Val Val Asn Gly Phe Ser Thr Lys
145 150 155 160
Val Arg Val Val Asp Ile Pro Lys Leu Lys Gln Ile Ala Gly Val Lys
165 170 175
Thr Val Thr Leu Ala Lys Val Tyr Tyr Pro Thr Asp Ala Lys Ala Asn
180 185 190
Ser Met Ala Asn Val Gln Ala Val Trp Ser Asn Tyr Lys Tyr Lys Gly
195 200 205
Glu Gly Thr Val Val Ser Val Ile Asp Ser Gly Ile Asp Pro Thr His
210 215 220
Lys Asp Met Arg Leu Ser Asp Asp Lys Asp Val Lys Leu Thr Lys Ser
225 230 235 240
Asp Val Glu Lys Phe Thr Asp Thr Val Lys His Gly Arg Tyr Phe Asn
245 250 255
Ser Lys Val Pro Tyr Gly Phe Asn Tyr Ala Asp Asn Asn Asp Thr Ile
260 265 270
Thr Asp Asp Lys Val Asp Glu Gln His Gly Met His Val Ala Gly Ile
275 280 285
Ile Gly Ala Asn Gly Thr Gly Asp Asp Pro Ala Lys Ser Val Val Gly
290 295 300
Val Ala Pro Glu Ala Gln Leu Leu Ala Met Lys Val Phe Ser Asn Ser
305 310 315 320
Asp Thr Ser Ala Lys Thr Gly Ser Ala Thr Val Val Ser Ala Ile Glu
325 330 335
Asp Ser Ala Lys Ile Gly Ala Asp Val Leu Asn Met Ser Leu Gly Ser
340 345 350
Asn Ser Gly Asn Gln Thr Leu Glu Asp Pro Glu Leu Ala Ala Val Gln
355 360 365
Asn Ala Asn Glu Ser Gly Thr Ala Ala Val Ile Ser Ala Gly Asn Ser
370 375 380
Gly Thr Ser Gly Ser Ala Thr Glu Gly Val Asn Lys Asp Tyr Tyr Gly
385 390 395 400
Leu Gln Asp Asn Glu Met Val Gly Ser Pro Gly Thr Ser Arg Gly Ala
405 410 415
Thr Thr Val Ala Ser Ala Glu Asn Thr Asp Val Ile Thr Gln Ala Val
420 425 430
Thr Ile Thr Asp Gly Thr Gly Leu Gln Leu Gly Pro Glu Thr Ile Gln
435 440 445
Leu Ser Ser His Asp Phe Thr Gly Ser Phe Asp Gln Lys Lys Phe Tyr
450 455 460
Ile Val Lys Asp Ala Ser Gly Asn Leu Ser Lys Gly Ala Leu Ala As
465 470 475 480
Tyr Thr Ala Asp Ala Lys Gly Lys Ile Ala Ile Val Lys Arg Gly Glu
485 490 495
Phe Ser Phe Asp Asp Lys Gln Lys Tyr Ala Gln Ala Ala Gly Ala Ala
500 505 510
Gly Leu Ile Ile Val Asn Thr Asp Gly Thr Ala Thr Pro Met Thr Ser
515 520 525
Ile Ala Leu Thr Thr Thr Phe Pro Thr Phe Gly Leu Ser Ser Val Thr
530 535 540
Gly Gln Lys Leu Val Asp Trp Val Thr Ala His Pro Asp Asp Ser Leu
545 550 555 560
Gly Val Lys Ile Thr Leu Ala Met Leu Pro Asn Gln Lys Tyr Thr Glu
565 570 575
Asp Lys Met Ser Asp Phe Thr Ser Tyr Gly Pro Val Ser Asn Leu Ser
580 585 590
Phe Lys Pro Asp Ile Thr Ala Pro Gly Gly Asn Ile Trp Ser Thr Gln
595 600 605
Asn Asn Asn Gly Tyr Thr Asn Met Ser Gly Thr Ser Met Ala Ser Pro
610 615 620
Phe Ile Ala Gly Ser Gln Ala Leu Leu Lys Gln Ala Leu Asn Asn Lys
625 630 635 640
Asn Asn Pro Phe Tyr Ala Tyr Tyr Lys Gln Leu Lys Gly Thr Ala Leu
645 650 655
Thr Asp Phe Leu Lys Thr Val Glu Met Asn Thr Ala Gln Pro Ile Asn
660 665 670
Asp Ile Asn Tyr Asn Asn Val Ile Val Ser Pro Arg Arg Gln Gly Ala
675 680 685
Gly Leu Val Asp Val Lys Ala Ala Ile Asp Ala Leu Glu Lys Asn Pro
690 695 700
Ser Thr Val Val Ala Glu Asn Gly Tyr Pro Ala Val Glu Leu Lys Asp
705 710 715 720
Phe Thr Ser Thr Asp Lys Thr Phe Lys Leu Thr Phe Thr Asn Arg Thr
725 730 735
Thr His Glu Leu Thr Tyr Gln Met Asp Ser Asn Thr Asp Thr Asn Ala
740 745 750
ValTyr Thr Ser Ala Thr Asp Pro Asn Ser Gly Val Leu Tyr Asp Lys
755 760 765
Lys Ile Asp Gly Ala Ala Ile Lys Ala Gly Ser Asn Ile Thr Val Pro
770 775 780
Ala Gly Lys Thr Ala Gln Ile Glu Phe Thr Leu Ser Leu Pro Lys Ser
785 790 795 800
Phe Asp Gln Gln Gln Phe Val Glu Gly Phe Leu Asn Phe Lys Gly Ser
805 810 815
Asp Gly Ser Arg Leu Asn Leu Pro Tyr Met Gly Phe Phe Gly Asp Trp
820 825 830
Asn Asp Gly Lys Ile Val Asp Ser Leu Asn Gly Ile Thr Tyr Ser Pro
835 840 845
Ala Gly Gly Asn Phe Gly Thr Val Pro Leu Leu Lys Asn Lys Asn Thr
850 855 860
Gly Thr Gln Tyr Tyr Gly Gly Met Val Thr Asp Ala Asp Gly Asn Lys
865 870 875 880
Thr Val Asp Asp Gln Ala Ile Ala Phe Ser Ser Asp Lys Asn Ala Leu
885 890 895
Tyr Asn Asp Ile Ser Met Lys Tyr Tyr Leu Leu Arg Asn Ile Ser Asn
900 905 910
Val Gln Val Asp Ile Leu Asp Gly Gln Gly Asn Lys Val Thr Thr Leu
915 920 925
Ser Ser Ser Thr Asn Arg Lys Lys Thr Tyr Tyr Asn Ala His Ser Gln
930 935 940
Gln Tyr Ile Tyr Tyr Asn Ala Pro Ala Trp Asp Gly Thr Tyr Tyr Asp
945 950 955 960
Gln Arg Asp Gly Asn Ile Lys Thr Ala Asp Asp Gly Ser Tyr Thr Tyr
965 970 975
Arg Ile Ser Gly Val Pro Glu Gly Gly Asp Lys Arg Gln Val Phe Asp
980 985 990
Val Pro Phe Lys Leu Asp Ser Lys Ala Pro Thr Val Arg His Val Ala
995 1000 1005
Leu Ser Ala Lys Thr Glu Asn Gly Lys Thr Gln Tyr Tyr Leu Thr Ala
1010 1015 1020
Glu Ala Lys Asp Asp Leu Ser Gly Leu Asp Ala Thr Lys Ser Val Lys
1025 1030 1035 1040
Thr Glu Ile Asn Glu Val Thr Asn Leu Asp Ala Thr Phe Thr Asp Ala
1045 1050 1055
Gly Thr Thr Ala Asp Gly Tyr Thr Lys Ile Glu Thr Pro Leu Ser Asp
1060 1065 1070
Glu Gln Ala Gln Ala Leu Gly Asn Gly Asp Asn Ser Ala Glu Leu Tyr
1075 1080 1085
Leu Thr Asp Asn Ala Ser Asn Ala Thr Asp Gln Asp Ala Ser Val Gln
1090 1095 1100
Lys Pro Gly Ser Thr Ser Phe Asp Leu Ile Val Asn Gly Gly Gly Ile
1105 1110 1115 1120
Pro Asp Lys Ile Ser Ser Thr Thr Thr Gly Tyr Glu Ala Asn Thr Gln
1125 1130 1135
Gly Gly Gly Thr Tyr Thr Phe Ser Gly Thr Tyr Pro Ala Ala Val Asp
1140 1145 1150
Gly Thr Tyr Thr Asp Ala Gln Gly Lys Lys His Asp Leu Asn Thr Thr
1155 1160 1165
Tyr Asp Ala Ala Thr Asn Ser Phe Thr Ala Ser Met Pro Val Thr Asn
1170 1175 1180
Ala Asp Tyr Ala Ala Gln Val Asp Leu Tyr Ala Asp Lys Ala His Thr
1185 1190 1195 1200
Gln Leu Leu Lys His Phe Asp Thr Lys Val Arg Leu Met Ala Pro Thr
1205 1210 1215
Phe Thr Asp Leu Lys Phe Asn Asn Gly Ser Asp Gln Thr Ser Glu Ala
1220 1225 1230
Thr Ile Lys Val Thr Gly Thr Val Ser Ala Asp Thr Lys Thr Val Asn
1235 1240 1245
Val Gly His Thr Val Ala Ala Leu Asp Ala Gln His His Phe Ser Val
1250 1255 1260
Asp Val Pro Val Asn Tyr Gly Asp Asn Thr Ile Lys Val Thr Ala Thr
1265 1270 1275 1280
Asp Lys Asp Gly Asn Thr Thr Thr Glu Gln Lys Thr Ile Thr Ser Ser
1285 1290 1295
Tyr Asp Pro Asp Met Leu Lys Lys Ser Val Thr Phe Asp Gln Gly Val
1300 1305 1310
Lys Phe Gly Thr Asn Lys Phe Asn Ala Thr Ser Ala Lys Phe Tyr Asp
1315 1320 1325
Pro Lys Thr Gly Ile Ala Thr Ile Thr Gly Lys Val Lys His Pro Thr
1330 1335 1340
Thr Thr Leu Gln Val Asp Gly Lys Gln Ile Pro Ile Lys Asp Asp Leu
1345 1350 1355 1360
Thr Phe Ser Phe Thr Leu Asp Leu Gly Thr Leu Gly Gln Lys Pro Phe
1365 1370 1375
Gly Val Val Val Gly Asp Thr Thr Gln Asn Lys Thr Phe Gln Glu Ala
1380 1385 1390
Leu Ser Phe Ile Leu Asp Ala Val Ala Pro Thr Leu Ser Leu Asp Ser
1395 1400 1405
Ser Thr Asp Ala Pro Val Tyr Thr Asn Asp Pro Asn Phe Gln Ile Thr
1410 1415 1420
Gly Thr Ala Thr Asp Asn Ala Gln Tyr Leu Ser Leu Ser Ile Asn Gly
1425 1430 1435 1440
Ser Ser Val Ala Ser Gln Tyr Glu Asp Ile Asn Ile Asn Ser Gly Lys
1445 1450 1455
Pro Gly His Met Ala Ile Asp Gln Pro Val Lys Leu Leu Glu Gly Lys
1460 1465 1470
Asn Val Leu Thr Val Ala Val Thr Asp Ser Glu Asp Asn Thr Thr Thr
1475 1480 1485
Lys Asn Ile Thr Val Tyr Tyr Glu Pro Lys Lys Thr Leu Ala Ala Pro
1490 1495 1500
Thr Val Thr Pro Ser Thr Thr Glu Pro Ala Gln Thr Val Thr Leu Thr
1505 1510 1515 1520
Ala Asn Ala Ala Ala Thr Gly Glu Thr Val Gln Tyr Ser Ala Asp Gly
1525 1530 1535
Gly Lys Thr Tyr Gln Asp Val Pro Ala Ala Gly Val Thr Ile Thr Ala
1540 1545 1550
Asn Gly Thr Phe Lys Phe Lys Ser Thr Asp Leu Tyr Gly Asn Glu Ser
1555 1560 1565
Pro Ala Val Asp Tyr Val Val Thr Asn Ile Lys Ala Asp Asp Pro Ala
1570 1575 1580
Gln Leu Gln Ala Ala Lys Gln Glu Leu Thr Asn Leu Ile Ala Ser Ala
1585 1590 1595 1600
Lys Thr Leu Ser Ala Ser Gly Lys Tyr Asp Asp Ala Thr Thr Thr Ala
1605 1610 1615
Leu Ala Ala Ala Thr Gln Lys Ala Gln Thr Ala Leu Asp Gln Thr Asn
1620 1625 1630
Ala Ser Val Asp Ser Leu Thr Gly Ala Asn Arg Asp Leu Gln Thr Ala
1635 1640 1645
Ile Asn Gln Leu Ala Ala Lys Leu Pro Ala Asp Lys Lys Thr Ser Leu
1650 1655 1660
Leu Asn Gln Leu Gln Ser Val Lys Asp Ala Leu Gly Thr Asp Leu Gly
1665 1670 1675 1680
Asn Gln Thr Asp Pro Ser Thr Gly Lys Thr Phe Thr Ala Ala Leu Asp
1685 1690 1695
Asp Leu Val Ala Gln Ala Gln Ala Gly Thr Gln Thr Asp Asp Gln Leu
1700 1705 1710
Gln Ala Thr Leu Ala Lys Ile Leu Asp Glu Val Leu Ala Lys Leu Ala
1715 1720 1725
Glu Gly Ile Lys Ala Ala Thr Pro Ala Glu Val Gly Asn Ala Lys Asp
1730 1735 1740
Ala Ala Thr Gly Lys Thr Trp Tyr Ala Asp Ile Ala Asp Thr Leu Thr
1745 1750 1755 1760
Ser Gly Gln Ala Ser Ala Asp Ala Ser Asp Lys Leu Ala His Leu Gln
1765 1770 1775
Ala Leu Gln Ser Leu Lys Thr Lys Val Ala Ala Ala Val Glu Ala Ala
1780 1785 1790
Lys Thr Val Gly Lys Gly Asp Gly Thr Thr Gly Thr Ser Asp Lys Gly
1795 1800 1805
Gly Gly Gln Gly Thr Pro Ala Pro Ala Pro Gly Asp Thr Gly Lys Asp
1810 1815 1820
Lys Gly Asp Glu Gly Ser Gln Pro Ser Ser Gly Gly Asn Ile Pro Thr
1825 1830 1835 1840
Lys Pro Ala Thr Thr Thr Ser Thr Thr Thr Asp Asp Thr Thr Asp Arg
1845 1850 1855
Asn Gly Gln Leu Thr Ser Gly Lys Gly Ala Leu Pro Lys Thr Gly Glu
1860 1865 1870
Thr Thr Glu Arg Pro Ala Phe Gly Phe Leu Gly Val Ile Val ValSer
1875 1880 1885
Leu Met Gly Val Leu Gly Leu Lys Arg Lys Gln Arg Glu Glu
1890 1895 1900
<210>322
<211>8115
<212>DNA
<213>Lactococcus lactis
<220>
<223>PIII-type proteinase precursor
(Lactocepin)S08.116
<400>322
ttttgaattg tttgccttta aaacgattca ttactctgtc ctctctgtct tttttctcaa 60
ttttacacta aaatagattt tttggaaaac tttgcaacag aaccagatct atcaactcta 120
cttgacgaag agccaatttc taaaagaact ggctgttatt tcatcttact cggttaagcc 180
ttaattcgtt gtcgcaagct tcttataact atctagcgca tccgcaagat ccttgtcttt 240
aatcgtcaca tgctgattct tcaatacctg actgataacg cgttgcatga tgcttgaatc 300
gcgagaccat ttagcgtaaa cgctggcagt tagcgccttt ttactgctag taaaggtgcc 360
tttggcggga tggttaatca ttttaataac ttcatacccg tctgtcactt tgactggggt 420
ctgcgtgtag tcaccatttt ttaatttgta ggcagcatcc ttaaatgtgg catcgagcgt 480
tttattgttt aattcaaaac taatcttccc gccgttatct ttagtcgcag tatcaatgga 540
atcagttttc gcaagcatgg caaaatcctt gccagctgct aaatcactga taacttgctt 600
agcagtgtcc tcgtcgctag ttagaatatg ttggacagtc actttgggct gataggtctt 660
ccaagcggcc ttgagctggc tttcagaaac cttttttaac tttttcagtg caacttcact 720
taaaaagttg gttcgtaggc tttccttgaa gctactgcga ctgaaaccgt tttgacttaa 780
gaaagcatcg aaattttcgc cgtattgttg tttgtagcta tcataggcat cattaactgt 840
tttagtgcta accgattttc cataggcatg attcaatgca cgataaatga gcatgttagc 900
aagcatggtc tttgtcgttg gtgactgttt gagttccttg tacaagctac tttcagtcac 960
cttgccacct gaataggtcg caactgtttg gtcggtctga tttgactgac aaccgcttag 1020
cagcagtaaa gcagttgcgg tacttgccaa taatacttta aggcgcattt ttttcttcat 1080
cgattcggtc tcctctgaaa tgcttacagt aaacgttgtg tatttattct agcgatggca 1140
attgagaatt tcaattcaaa tactcatttt ctaataagtg ctaaaaattt caaaacatct 1200
tagtctgtaa acggctaaat aataacgcta aaagttaatt tacagataaa aaaattaata 1260
agaagattaa aattttcgtt gaatttgttc ttcaatagta tataatataa tagtatataa 1320
attatattat ataatataat cttaactaca tcaagcgtag gctttgattt ggttatgaaa 1380
acttttggaa agtggaggat attggatgca aaggaaaaag aaagggctat cgatcttgtt 1440
acagtaaacg ttgtgtattt attctagcga tggcaattga gaatttcaat tcaaatactc 1500
attttctaat aaytgctaaa aatttcaaaa catctatagt ctgtaaacgg ctaaataata 1560
acgctaaaag ttaatttaca gataaaaaaa ttaatagaag attaaaattt tcgttgaatt 1620
tgttcttcaa tagtatataa tataatagta tataatatta tattatataa tataatctta 1680
actacatcaa gcgtaggctt tgatttggtt atgaaacttt tggaaagtgg aggatattgg 1740
atgcaaagga aaaagaaagg gctatcgatc ttgttagccg gtacagtcgc tttaggggcg 1800
ctggctgtct tgccagtcgg cgaaatccaa gcaaaggcgg ctatctcgca gcaaactaag 1860
ggatcatcac tcgcaaatac ggtgacggct gcgactgcta agcaagcggc cactgacaca 1920
actgcagcga caacgaatca agcgattgct acacagttgg cggctaaagg tattgattac 1980
aataagctga ataaagttca gcagcaagat atttatgttg acgtcattgt tcaaatgagc 2040
gcagcgcctg cctctgaaaa cggcatttta agaactgatt actccagcac ggcggagatt 2100
cagcaggaga ccaataaagt gatcgcggct caggcaagcg ttaaagcagc tgttgaacaa 2160
gtcacccaac aaactgccgg tgaaagttat ggctatgtcg ttaacggctt ttcaactaaa 2220
gttagggttg ttgatatccc taaactgaaa caaattgccg gagttaaaac agtcacattg 2280
gcgaaagttt attatccgac tgatgctaag gcaaactcga tggcgaatgt gcaagccgta 2340
tggtccaatt acaaatataa aggtgaaggc acagttgtct cggttattga cagtggcatt 2400
gatccaacac ataaagacat gcggctaagc gatgataaag acgttaaact aaccaaatct 2460
gatgttgaaa aattcactga taccgttaag catggccgct attttaattc aaaagtgcca 2520
tatgggttta attacgctga taataacgac accattacag atgataaggt tgacgaacaa 2580
cacggcatgc atgttgctgg gatcatcggt gctaacggga caggtgacga tccagccaag 2640
tctgttgtcg gagttgcacc agaagcacag ctactggcaa tgaaagtttt cagcaactct 2700
gacacttctg caaaaaccgg gtcagctacc gtggtttctg ccattgaaga ctcggcaaaa 2760
atcggtgccg atgtcctcaa catgtcctta ggatctaatt caggcaacca aaccttggag 2820
gatccagaac ttgctgcggt gcaaaatgct aacgaatcag gaacagccgc cgtcatttct 2880
gctgggaact caggaacatc cggttcagca actgaaggcg tcaacaaaga ttattacggt 2940
ttgcaagaca atgaaatggt gggatcgcca gggacatcac gaggagcgac cacagttgct 3000
tccgctgaaa acacggatgt catcactcag gcagtgacca ttacagatgg tacaggttta 3060
cagcttggac cggaaaccat tcagctttca agccacgatt tcactggtag ctttgaccaa 3120
aagaagtttt atattgttaa agatgctagt ggcaacctca gcaaaggggc attagccgac 3180
tatactgctg acgctaaagg caaaattgcc atcgttaaac gtggcgaatt tagctttgat 3240
gacaaacaaa aatacgccca agccgctggt gctgctggct tgatcattgt caacaccgat 3300
ggcacagcaa caccgatgac ttctattgcg ttaaccacca ccttcccaac atttgggctc 3360
tccagtgtaa ccggtcaaaa gctggttgac tgggtcacag cacacccgga tgatagtctc 3420
ggtgtcaaga ttaccctggc gatgttacca aatcagaaat atactgaaga caagatgtct 3480
gacttcacat cctatgggcc agtttccaat ctttccttca aaccagatat taccgcacca 3540
ggcggcaaca tctggtcaac gcaaaacaac aatggctaca caaatatgtc tggtacgtca 3600
atggcctcgc catttattgc cggttcacaa gcattgttga aacaagcatt gaataacaaa 3660
aacaacccat tttatgctta ctacaaacaa cttaaaggga cagcgctcac cgattttctt 3720
aagacagttg agatgaatac tgcccagcca atcaacgata ttaactacaa taatgttatc 3780
gtatcgccgc ggcggcaagg ggccggtctg gttgatgtga aggcagccat tgatgcatta 3840
gaaaagaatc cgtcaacggt tgtcgccgaa aacggctacc cggcagttga attgaaagac 3900
ttcacgagta cggacaagac ctttaaactc accttcacga atcgcacgac ccatgaacta 3960
acctatcaaa tggacagtaa tacggatact aatgccgttt atacatcagc gactgaccct 4020
aattctgggg ttttgtatga caagaagatt gatggagcag ccattaaagc tggcagtaac 4080
ataactgtgc ctgctgggaa aacggcgcag attgaattca cactatcttt gccgaagtct 4140
tttgaccaac agcaatttgt tgaaggtttt ctgaacttta agggtagcga tggatcgcgc 4200
ttgaacttgc catacatggg cttttttggt gactggaatg acggtaagat tgtcgatagt 4260
ctcaatggga tcacttatag tcctgctggt ggtaattttg gcaccgtgcc actattgaaa 4320
aacaaaaata caggcactca atattatggc ggcatggtca cagacgctga tggcaacaag 4380
acagttgacg atcaggcgat tgctttttcg agtgacaaga atgccttata taatgacatc 4440
agcatgaagt attatctatt gcgcaatatc agcaacgtcc aagttgatat tcttgatggt 4500
cagggcaata aagttacgac tctcagcagt tccaccaatc ggaagaagac ctattataat 4560
gctcattcgc agcagtacat ctactacaat gctccagcgt gggatggcac ctattatgat 4620
caacgtgatg gcaacatcaa gacggctgat gatggcagtt atacttatcg tatttccggt 4680
gtaccggaag gcggcgacaa acgtcaagtg tttgatgtgc ctttcaagct cgactctaag 4740
gcgccgacag ttcgtcatgt cgctttgtca gccaaaacgg aaaatgggaa aacccagtat 4800
tatttgacag ctgaagccaa ggatgatttg agtggtcttg atgccaccaa gagcgttaaa 4860
actgaaatta atgaagtgac gaatcttgat gctaccttta ccgatgctgg gacaacggct 4920
gatgggtaca ccaaaattga aacgccatta tctgatgaac aggcccaagc acttggcaat 4980
ggcgacaatt cggctgagct gtacttgact gataatgcat ccaatgccac tgatcaagat 5040
gccagcgttc agaagccggg gtctacatcg tttgatttaa ttgtgaacgg cggcggtatc 5100
ccagacaaga tctcaagtac cacaaccggc tacgaagcca atactcaagg tggcgggacg 5160
tatacgttta gtggaacgta tccagcagcg gttgacggta cttacactga tgcacaagga 5220
aagaaacatg atttgaacac aacctacgat gctgcgacta acagtttcac tgcctcaatg 5280
ccggtcacga atgctgatta cgccgcgcaa gtggatctat atgccgataa ggcgcatacc 5340
cagttgctta aacattttga caccaaagtt cggctgatgg cgccaacctt tactgatttg 5400
aaattcaaca atggctcgga tcagacctct gaagcgacca tcaaggttac agggacggtt 5460
agtgctgaca ccaagacagt taatgttggc cacaccgtag cagcacttga tgcacaacat 5520
cactttagtg ttgatgtacc ggttaattat ggtgacaata ccatcaaggt gaccgccacc 5580
gacaaagatg gcaacaccac gacggagcaa aagacgatta cctcgtctta tgatcctgat 5640
atgttgaaga agtctgtgac gttcgatcaa ggtgtgaaat ttggtaccaa taaattcaat 5700
gccacctcgg ctaagttcta tgaccctaag accgggattg cgacgattac tggtaaggtc 5760
aagcacccaa cgacaacgtt gcaggttgat ggtaagcaaa ttccaatcaa ggatgatctg 5820
actttcagtt tcactttaga tttaggtact cttggacaaa aaccgtttgg ggttgttgtg 5880
ggtgacacca ctcaaaacaa aaccttccaa gaagcgttgt ccttcatttt ggatgcagtg 5940
gctccaacat tgtcattgga tagctcgaca gatgcaccgg tttataccaa cgatccaaac 6000
ttccagatta ccggaacggc cactgacaat gcgcaatatc tgagtctgtc aattaacggc 6060
agttctgtcg ccagccaata cgaagacatc aacatcaata gcggcaaacc aggtcatatg 6120
gctattgatc agcccgttaa attgctcgaa ggcaaaaacg tgctgactgt tgctgttaca 6180
gatagcgaag acaacaccac gaccaagaac atcacagttt actacgaacc aaagaaaaca 6240
ctggcagcac caactgtgac gccaagtacc actgaaccag cccaaacggt gactctgacg 6300
gcaaacgctg ccgcaacggg cgaaacggtt cagtatagtg ctgatggtgg caagacatat 6360
caggatgttc cggcagccgg tgtcacgatc acggcaaatg gcaccttcaa gtttaagtcg 6420
actgatttat acggtaatga atcaccagcc gtcgactatg ttgtcaccaa tatcaaggcc 6480
gatgatcctg cacaattgca ggcagctaag caggaactga ctaatctgat tgcttccgcc 6540
aaaacgctaa gtgccagcgg taagtatgat gatgccacaa cgactgcttt agcagcggca 6600
acgcagaagg cacaaacggc gcttgatcag acgaacgcct cagttgattc acttactggt 6660
gccaatcgag acctgcaaac tgcgatcaat caattagctg ccaagttacc tgcagataag 6720
aagacttcgc tgcttaacca gttgcaatct gtgaaggatg cgctgggaac ggacttgggc 6780
aatcaaactg atccaagcac tggcaaaaca tttacggcag cgttagacga cctagtggca 6840
caagctcaag caggcacgca aacggacgac cagcttcaag caactcttgc caagatactt 6900
gatgaagtat tagcaaaact tgcggagggt attaaagcgg caacaccggc tgaggttggc 6960
aatgctaaag atgctgcaac tggcaaaact tggtatgccg acattgctga cacattgacg 7020
tctggtcaag ccagtgctga tgcgtctgac aagcttgcac atttacaagc tttgcaaagt 7080
ctgaaaacga aggtggcagc tgccgttgaa gcggccaaga cagttggtaa aggcgacggt 7140
acaaccggta ctagcgacaa aggcggcggt caaggtaccc cggcgcccgc tccaggcgac 7200
acaggtaagg acaaaggaga tgagggcagc cagcctagtt ctggcggtaa tatcccaaca 7260
aagccagcca caacgacgtc aacgaccacg gatgatacga ctgatcgtaa tggtcaactt 7320
acatccggta ctagcgacaa aggcggcggt caaggtaccc cggcgcccgc tccaggcgac 7380
ataggtaagg acaaaggcga tgagggcagc cagcctagtt ctggcggtaa tatcccaaca 7440
aatccagcca caacgacgtc aacgaccacg gatgatacga ctgatcgtaa tggtcaactt 7500
acatccggta agggagcatt acccaagaca ggagagacaa ctgagcggcc agcgtttggc 7560
ttcttgggtg tcattgtggt cagtctgatg ggggtattag gattgaaacg gaaacaacgt 7620
gaagaatagc ctgtccatgc gttgcgttta gggcagcagc gtatcgacaa aaagaaaagg 7680
gcgctacgat atttggagtt gaggttcaaa gtcaaatggt actgatgacc ggtaaaattt 7740
aatattttga accttgctta ggcagctgac ttcacattgt tgagatcagc tgccttttgc 7800
ttatagttca ttgagtagaa acggttctgt tgcgaagttt gaaaatcaaa cgcgcccgct 7860
ctgaactcca aatatcttag gctgttattc ccattaatac cttgatttca gtagacaccg 7920
aaaagccgaa gagcgttcca tttcttcggt tatttttata tattcctcga agggtctcca 7980
tgcccttaat cgtggaagag gctgtacgga gactttgata aaatttaatc cgtcgtttaa 8040
taggtcgatg gtcttgttct attaaattgt taagatactt cacagttcgg tgctctgtct 8100
tagtatataa accca 8115
<210>323
<211>1101
<212>PRT
<213>Drosophila melanogaster
<220>
<223>Furin-like protease 1，isoform 1-CRR precursor
(Furin-1) S08.048
<400>323
Met Lys Asn Asp Val Val Arg Trp Ser Arg Gln Pro Thr Ser Asn Thr
1 5 10 15
Thr Asn Ser Ser Ser Ser Ser Arg Ser Asp Ser Asn Ser Thr His Lys
20 25 30
His Arg Ser Lys Ser Asn Lys Leu Asn Ala Arg Gln Leu Gly Ser Asn
35 40 45
Ala Ala Arg Ser Cys Gln Gln Arg Ser Ser Val Ala Thr Thr Leu Glu
50 55 60
Asp Glu Gln Gln Thr Ile Ile Glu Cys Asp Ile Gly Asn Phe Asn Phe
65 70 75 80
Asp Cys Asn Leu Phe Lys Thr Ser Phe Leu Thr Gln His Lys Gln Lys
85 90 95
Arg Ser Gly Lys Ser Ser Ser Lys Ser Lys Ser Asn Arg Ser Arg Pro
100 105 110
Leu Ala Lys Thr Lys Ala Val Phe Leu Leu Ala Leu Gln Phe Ser Ala
115 120 125
Val Val Phe Leu Cys Asn Ile Asn Val Gly Phe Val Ala Gly Ser Val
130 135 140
Ala Thr Ala Ala Ser Ser Ala Gly Gly Ser Ser Pro Ala Ala Pro Ser
145 150 155 160
Ser Ala Pro Ser Ser Pro Pro Thr Val Ala Val Pro Pro Pro Pro Pro
165 170 175
Pro Ser Ser Ala Leu Lys Val Asp Pro Asn Gly Gln Ser Pro Val Leu
180 185 190
Pro Pro Tyr Val Leu Asp Tyr Glu Thr Gly Gly Lys Ala Lys Leu Thr
195 200 205
Pro Asn Asn Gly Lys Phe Gly Gln Ser Gly Ser Ser Gly Ser Asn Asn
210 215 220
Asn His Ile Val Gly His Tyr Thr His Thr Trp Ala Val His Ile Pro
225 230 235 240
Asn Gly Asp Asn Gly Met Ala Asp Ala Val Ala Lys Asp His Gly Phe
245 250 255
Val Asn Leu Gly Lys Ile Phe Asp Asp His Tyr His Phe Ala His His
260 265 270
Lys Val Ser Lys Arg Ser Leu Ser Pro Ala Thr His His Gln Thr Arg
275 280 285
Leu Asp Asp Asp Asp Arg Val His Trp Ala Lys Gln Gln Arg Ala Lys
290 295 300
Ser Arg Ser Lys Arg Asp Phe Ile Arg Met Arg Pro Ser Arg Thr Ser
305 310 315 320
Ser Arg Ala Met Ser Met Val Asp Ala Met Ser Phe Asn Asp Ser Lys
325 330 335
Trp Pro Gln Met Trp Tyr Leu Asn Arg Gly Gly Gly Leu Asp Met Asn
340 345 350
Val Ile Pro Ala Trp Lys Met Gly Ile Thr Gly Lys Gly Val Val Val
355 360 365
Thr Ile Leu Asp Asp Gly Leu Glu Ser Asp His Pro Asp Ile Gln Asp
370 375 380
Asn Tyr Asp Pro Lys Ala Ser Tyr Asp Val Asn Ser His Asp Asp Asp
385 390 395 400
Pro Met Pro His Tyr Asp Met Thr Asp Ser Asn Arg His Gly Thr Arg
405 410 415
Cys Ala Gly Glu Val Ala Ala Thr Ala Asn Asn Ser Phe Cys Ala Val
420 425 430
Gly Ile Ala Tyr Gly Ala Ser Val Gly Gly Val Arg Met Leu Asp Gly
435 440 445
Asp Val Thr Asp Ala Val Glu Ala Arg Ser Leu Ser Leu Asn Pro Gln
450 455 460
His Ile Asp Ile Tyr Ser Ala Ser Trp Gly Pro Asp Asp Asp Gly Lys
465 470 475 480
Thr Val Asp Gly Pro Gly Glu Leu Ala Ser Arg Ala Phe Ile Glu Gly
485 490 495
Thr Thr Lys Gly Arg Gly Gly Lys Gly Ser Ile Phe Ile Trp Ala Ser
500 505 510
Gly Asn Gly Gly Arg Glu Gln Asp Asn Cys Asn Cys Asp Gly Tyr Thr
515 520 525
Asn Ser Ile Trp Thr Leu Ser Ile Ser Ser Ala Thr Glu Glu Gly His
530 535 540
Val Pro Trp Tyr Ser Glu Lys Cys Ser Ser Thr Leu Ala Thr Thr Tyr
545 550 555 560
Ser Ser Gly Gly Gln Gly Glu Lys Gln Val Val Thr Thr Asp Leu His
565 570 575
His Ser Cys Thr Val Ser His Thr Gly Thr Ser Ala Ser Ala Pro Leu
580 585 590
Ala Ala Gly Ile Ala Ala Leu Val Leu Gln Ser Asn Gln Asn Leu Thr
595 600 605
Trp Arg Asp Leu Gln His Ile Val Val Arg Thr Ala Lys Pro Ala Asn
610 615 620
Leu Lys Asp Pro Ser Trp Ser Arg Asn Gly Val Gly Arg Arg Val Ser
625 630 635 640
His Ser Phe Gly Tyr Gly Leu Met Asp Ala Ala Glu Met Val Arg Val
645 650 655
Ala Arg Asn Trp Lys Ala Val Pro Glu Gln Gln Arg Cys Glu Ile Asn
660 665 670
Ala Pro His Val Asp Lys Val Ile Pro Pro Arg Thr His Ile Thr Leu
675 680 685
Gln Leu Thr Val Asn His Cys Arg Ser Val Asn Tyr Leu Glu His Val
690 695 700
Gln Ala Lys Ile Thr Leu Thr Ser Gln Arg Arg Gly Asp Ile Gln Leu
705 710 715 720
Phe Leu Arg Ser Pro Ala Asn Thr Ser Val Thr Leu Leu Thr Pro Arg
725 730 735
Ile His Asp Asn Ser Arg Ser Gly Phe Asn Gln Trp Pro Phe Met Ser
740 745 750
Val His Thr Trp Gly Glu Ser Pro Gln Gly Asn Trp Gln Leu Glu Ile
755 760 765
His Asn Glu Gly Arg Tyr Met Ala Gln Ile Thr Gln Trp Asp Met Ile
770 775 780
Phe Tyr Gly Thr Glu Thr Pro Ala Gln Pro Asp Asp Val Ala Asn Pro
785 790 795 800
Ser Gln Ser Asn Gln Phe Asn Leu Tyr Gly Asn Asp Met Ala His Asn
805 810 815
Asp Val Glu Tyr Asp Ser Thr Gly Gln Trp Arg Asn Met Gln Gln Val
820 825 830
Gly Glu Val Gly Met Thr Arg Asp His Ser Asn Thr Ala Ala Cys Leu
835 840 845
Lys Trp Ser Asp Arg Lys Cys Leu Glu Cys Asn Asp Ser Ala Tyr Met
850 855 860
Phe Glu Asp Gln Cys Tyr Asp Val Cys Pro Val His Thr Tyr Pro Leu
865 870 875 880
Asp Lys Phe Gln Ala Glu Glu Asp Glu Gln Asp Asp Glu Val Thr Arg
885 890 895
Gly Pro Val Asn Pro Tyr Ser Ser Ser Pro Met Asp His Ser Leu Leu
900 905 910
Met Ser Asn Ser Leu Asp Asp Lys Gln Asp Pro Leu Gln Ala Glu Asp
915 920 925
Arg Arg Arg Arg Ser Ser Leu Thr Gln Leu Val Glu Val Pro Ser Arg
930 935 940
Val Cys Ala Ala Cys Asp Arg Ser Cys Leu Glu Cys Tyr Gly Ala Leu
945 950 955 960
Ala Ser Gln Cys Ser Thr Cys Ser Pro Gly Ser Gln Leu Arg Lys Ile
965 970 975
Leu Asn Glu Thr Phe Cys Tyr Ala Tyr Val Val Arg Ser Thr Gly Met
980 985 990
Ala Ser Val Val Asp Ile Ser Lys Met Asp Asp Arg Asp Thr Gln Gln
995 1000 1005
Tyr Met Thr Gly Thr Thr Val Leu Leu Leu Val Ser Val Ile Phe Thr
1010 1015 1020
Leu Met Gly Val Ala Val Ala Gly Gly Ile Val Tyr His Arg Arg Ala
1025 1030 1035 1040
Met Ala Arg Ser Asn Glu Leu Tyr Ser Arg Val Ser Leu Val Pro Gly
1045 1050 1055
Asp Glu Ser Asp Ser Asp Glu Asp Glu Leu Phe Thr Ala His Phe Pro
1060 1065 1070
Ala Arg Lys Ser Gly Val Asn Ile Tyr Arg Asp Glu Ala Pro Ser Glu
1075 1080 1085
Lys Ile Phe Glu Glu Asp Glu Ile Ser His Leu Val Pro
1090 1095 1100
<210>324
<211>3781
<212>DNA
<213>Drosophila melanogaster
<220>
<223>Furin-like protease 1，isoform 1-CRR precursor
(Furin-1) S08.048
<400>324
gacaacaaca aatttaaatt gagattttcc atattatcaa aagttacagg cgcctatttt 60
gcagttatca gtctgaagtc ggtgtgaagt gctatcctgg acttttcgtt ttagtcaccc 120
cttggcctca tctaacatcc acttaattga aatagaacat ttgcatctca gcctcacaac 180
caatatcccg agcaacgtgc acaaaaacta ttggaaaaat cctaacccac aatgaaaaac 240
gacgtcgtgc gatggagcag gcagccaact agcaacacca ctaacagcag cagcagcagt 300
agaagcgata gcaacagcac tcataagcac agaagcaaaa gcaacaaatt aaatgctcga 360
caattagggt caaatgctgc cagaagttgt cagcaaagat cctctgtggc aacaacacta 420
gaagatgaac aacaaacaat cattgaatgt gatataggaa atttcaattt cgattgcaat 480
ttattcaaaa ctagtttttt aacgcaacac aaacagaaac gtagtggcaa gagcagcagc 540
aaaagtaaga gcaacagaag tagaccccta gcgaaaacga aagcggtgtt tctattagct 600
ctgcaattta gtgccgtagt ttttttatgt aatattaatg tcggtttcgt ggccggaagt 660
gtggcaactg cggcatcatc ggcaggcggc tcatcgccgg cagctccatc atctgcgccc 720
tcatccccgc ccacagttgc tgtaccaccg ccgccgccac cttcgtcggc actcaaagtg 780
gatccaaatg gtcagtcacc agtgctgccg ccctacgttc tcgattatga gacggggggc 840
aaggccaagc taacgccaaa caatggcaag ttcggccaat cgggcagttc cgggagcaat 900
aacaaccaca tcgtcggaca ctatacccac acctgggcgg tgcacatacc aaacggcgat 960
aatggcatgg ccgatgcggt tgccaaggat cacggattcg tcaatttggg caagatcttc 1020
gatgatcact accacttcgc acatcacaag gtctcgaagc ggtcgctctc ccccgccacg 1080
catcaccaaa ctcgcctgga tgacgacgat cgcgtccact gggcgaagca gcagcgggcc 1140
aagtcgcgat ccaaacggga ctttatccgc atgcgaccct cacggacctc ctcgcgagcc 1200
atgtcgatgg tggacgccat gtcctttaac gactccaagt ggccgcagat gtggtatctg 1260
aatcgtggtg gtggcctgga catgaatgtg atacccgcct ggaagatggg cataaccggc 1320
aagggcgtgg tggtgacaat tctggatgat ggcctggaat ccgatcatcc ggacatacag 1380
gataactacg atcccaaagc ctcgtacgat gtgaatagcc acgacgacga tccgatgccg 1440
cattacgata tgacggactc gaaccgccat ggaactcgct gtgccggcga ggtggcagcc 1500
accgccaaca attcgttctg cgcggtgggt attgcctacg gcgccagtgt gggcggagtc 1560
aggatgctgg acggagacgt cacggatgcg gttgaggcac ggtcgctgtc gctgaatccg 1620
cagcacattg acatatacag tgcctcctgg ggacccgatg acgatggcaa gacggtggac 1680
ggacccggcg aactggcatc gcgcgccttt atcgagggca caactaaggg acgcggcggc 1740
aagggcagca tcttcatatg ggcatcgggc aatggtgggc gggagcagga taactgcaac 1800
tgcgacggct acacgaactc catctggacg ctgtccatct ccagtgccac ggaggagggc 1860
catgtgccct ggtactcgga gaagtgcagc tccacgctgg ccaccaccta cagcagcggc 1920
gggcagggcg agaagcaggt ggtcaccacg gacctgcacc actcgtgcac tgtctcccac 1980
acgggcacct cggcgtcggc cccgctcgcc gctggcatag ccgccctggt gctgcagtcc 2040
aaccagaatc tcacctggcg cgatctgcag cacattgttg tgcgcaccgc caagccggcg 2100
aaccttaagg accccagctg gtcacgcaat ggggtggggc ggcgggtgag ccactccttt 2160
ggctacggat tgatggacgc cgccgagatg gtgcgcgtgg cccgcaactg gaaggcggtg 2220
ccggagcagc agcggtgcga gattaacgct ccccatgtcg acaaggtcat tccacctcgt 2280
acccatatca ccctgcaact gacggttaat cactgtcgat cggtcaatta cctggagcac 2340
gtccaggcca agattacgct aacgtcgcag agacgaggag acattcagct ctttttgagg 2400
tctcccgcaa acaccagtgt cacgctccta acgcctagga tacatgacaa ctctcgttcc 2460
ggattcaatc aatggccctt catgtctgtg cacacctggg gagagtcgcc gcaaggaaac 2520
tggcagctgg agatccacaa cgagggtcgc tatatggcac aaatcacaca atgggatatg 2580
atattctacg gcaccgaaac gcccgcccaa cccgatgacg tggccaatcc cagccagtcg 2640
aaccagttca atctgtacgg caacgatatg gcccacaatg acgtcgagta cgattccacc 2700
ggccagtgga ggaatatgca gcaggtgggc gaggtgggca tgacccgaga tcacagcaac 2760
accgccgcgt gccttaagtg gagcgatcgc aagtgcttag agtgcaatga ctccgcctat 2820
atgttcgagg accagtgcta cgatgtctgc ccggtgcaca catatccatt ggacaagttc 2880
caagcagagg aggatgagca ggatgacgag gtgacaaggg gtccggttaa tccctacagc 2940
agctccccca tggatcattc cctgttgatg tccaacagct tggatgacaa gcaagatcct 3000
ctgcaggcgg aggatcgacg tcgtcgtagt tcactcaccc aactggtgga ggtgccttct 3060
cgggtctgtg ccgcctgcga tcggagctgc ttggagtgct acggtgccct ggcctcccag 3120
tgcagcacct gctccccggg cagccaactg cgtaagatcc tgaacgagac cttctgctat 3180
gcctatgtgg tgcgtagcac gggaatggcc agtgtggtgg acatttccaa aatggacgat 3240
agagataccc agcaatatat gactggcaca attgtgctcc ttttggtttc ggtgattttc 3300
accctgatgg gcgtagcagt ggccggtggc atcgtgtatc atcgaagggc aatggctcga 3360
tccaacgaac tatactcccg agtttcccta gtccccggtg acgaaagtga ctctgacgag 3420
gatgaactgt ttacggccca cttcccagcg agaaagagtg gtgttaatat atatcgcgat 3480
gaagcgccca gcgaaaagat attcgaggaa gatgagatca gccacttggt accctaaata 3540
atcaaaacaa gttattagtt aggtcatgca tttagccatc ctcataacca ccagaaatat 3600
atctatatgt taagcaaccg acaaacgcat ttttggtcta tattttgtta cattcgacgg 3660
cattacagct cctttatttc aagccaaaat agattaaagt attaaatgta aaagaaaaaa 3720
aatcaaaact aattatttgt aatatttatg ccttaataaa atgattttaa ctgaaaaaaa 3780
a 3781
<210>325
<211>814
<212>PRT
<213>Saccharomyces cerevisiae
<220>
<223>Kexin precursor(KEX2 protease) S08.048
<400>325
Met Lys Val Arg Lys Tyr Ile Thr Leu Cys Phe Trp Trp Ala Phe Ser
1 5 10 15
Thr Ser Ala Leu Val Ser Ser Gln Gln Ile Pro Leu Lys Asp His Thr
20 25 30
Ser Arg Gln Tyr Phe Ala Val Glu Ser Asn Glu Thr Leu Ser Arg Leu
35 40 45
Glu Glu Met His Pro Asn Trp Lys Tyr Glu His Asp Val Arg Gly Leu
50 55 60
Pro Asn His Tyr Val Phe Ser Lys Glu Leu Leu Lys Leu Gly Lys Arg
65 70 75 80
Ser Ser Leu Glu Glu Leu Gln Gly Asp Asn Asn Asp His Ile Leu Ser
85 90 95
Val His Asp Leu Phe Pro Arg Asn Asp Leu Phe Lys Arg Leu Pro Val
100 105 110
Pro Ala Pro Pro Met Asp Ser Ser Leu Leu Pro Val Lys Glu Ala Glu
115 120 125
Asp Lys Leu Ser Ile Asn Asp Pro Leu Phe Glu Arg Gln Trp His Leu
130 135 140
Val Asn Pro Ser Phe Pro Gly Ser Asp Ile Asn Val Leu Asp Leu Trp
145 150 155 160
Tyr Asn Asn Ile Thr Gly Ala Gly Val Val Ala Ala Ile Val Asp Asp
165 170 175
Gly Leu Asp Tyr Glu Asn Glu Asp Leu Lys Asp Asn Phe Cys Ala Glu
180 185 190
Gly Ser Trp Asp Phe Asn Asp Asn Thr Asn Leu Pro Lys Pro Arg Leu
195 200 205
Ser Asp Asp Tyr His Gly Thr Arg Cys Ala Gly Glu Ile Ala Ala Lys
210 215 220
Lys Gly Asn Asn Phe Cys Gly Val Gly Val Gly Tyr Asn Ala Lys Ile
225 230 235 240
Ser Gly Ile Arg Ile Leu Ser Gly Asp Ile Thr Thr Glu Asp Glu Ala
245 250 255
Ala Ser Leu Ile Tyr Gly Leu Asp Val Asn Asp Ile Tyr Ser Cys Ser
260 265 270
Trp Gly Pro Ala Asp Asp Gly Arg His Leu Gln Gly Pro Ser Asp Leu
275 280 285
Val Lys Lys Ala Leu Val Lys Gly Val Thr Glu Gly Arg Asp Ser Lys
290 295 300
Gly Ala Ile Tyr Val Phe Ala Ser Gly Asn Gly Gly Thr Arg Gly Asp
305 310 315 320
Asn Cys Asn Tyr Asp Gly Tyr Thr Asn Ser Ile Tyr Ser Ile Thr Ile
325 330 335
Gly Ala Ile Asp His Lys Asp Leu His Pro Pro Tyr Ser Glu Gly Cys
340 345 350
Ser Ala Val Met Ala Val Thr Tyr Ser Ser Gly Ser Gly Glu Tyr Ile
355 360 365
His Ser Ser Asp Ile Asn Gly Arg Cys Ser Asn Ser His Gly Gly Thr
370 375 380
Ser Ala Ala Ala Pro Leu Ala Ala Gly Val Tyr Thr Leu Leu Leu Glu
385 390 395 400
Ala Asn Pro Asn Leu Thr Trp Arg Asp Val Gln Tyr Leu Ser Ile Leu
405 410 415
Ser Ala Val Gly Leu Glu Lys Asn Ala Asp Gly Asp Trp Arg Asp Ser
420 425 430
Ala Met Gly Lys Lys Tyr Ser His Arg Tyr Gly Phe Gly Lys Ile Asp
435 440 445
Ala His Lys Leu Ile Glu Met Ser Lys Thr Trp Glu Asn Val Asn Ala
450 455 460
Gln Thr Trp Phe Tyr Leu Pro Thr Leu Tyr Val Ser Gln Ser Thr Asn
465 470 475 480
Ser Thr Glu Glu Thr Leu Glu Ser Val Ile Thr Ile Ser Glu Lys Ser
485 490 495
Leu Gln Asp Ala Asn Phe Lys Arg Ile Glu His Val Thr Val Thr Val
500 505 510
Asp Ile Asp Thr Glu Ile Arg Gly Thr Thr Thr Val As Leu Ile Ser
515 520 525
Pro Ala Gly Ile Ile Ser Asn Leu Gly Val Val Arg Pro Arg Asp Val
530 535 540
Ser Ser Glu Gly Phe Lys Asp Trp Thr Phe Met Ser Val Ala His Trp
545 550 555 560
Gly Glu Asn Gly Val Gly Asp Trp Lys Ile Lys Val Lys Thr Thr Glu
565 570 575
Asn Gly His Arg Ile Asp Phe His Ser Trp Arg Leu Lys Leu Phe Gly
580 585 590
Glu Ser Ile Asp Ser Ser Lys Thr Glu Thr Phe Val Phe Gly Asn Asp
595 600 605
Lys Glu Glu Val Glu Pro Ala Ala Thr Glu Ser Thr Val Ser Gln Tyr
610 615 620
Ser Ala Ser Ser Thr Ser Ile Ser Ile Ser Ala Thr Ser Thr Ser Ser
625 630 635 640
Ile Ser Ile Gly Val Glu Thr Ser Ala Ile Pro Gln Thr Thr Thr Ala
645 650 655
Ser Thr Asp Pro Asp Ser Asp Pro Asn Thr Pro Lys Lys Leu Ser Ser
660 665 670
Pro Arg Gln Ala Met His Tyr Phe Leu Thr Ile Phe Leu Ile Gly Ala
675 680 685
Thr Phe Leu Val Leu Tyr Phe Met Phe Phe Met Lys Ser Arg Arg Arg
690 695 700
Ile Arg Arg Ser Arg Ala Glu Thr Tyr Glu Phe Asp Ile Ile Asp Thr
705 710 715 720
Asp Ser Glu Tyr Asp Ser Thr Leu Asp Asn Gly Thr Ser Gly Ile Thr
725 730 735
Glu Pro Glu Glu Val Glu Asp Phe Asp Phe Asp Leu Ser Asp Glu Asp
740 745 750
His Leu Ala Ser Leu Ser Ser Ser Glu Asn Gly Asp Ala Glu His Thr
755 760 765
Ile Asp Ser Val Leu Thr Asn Glu Asn Pro Phe Ser Asp Pro Ile Lys
770 775 780
Gln Lys Phe Pro Asn Asp Ala Asn Ala Glu Ser Ala Ser Asn Lys Leu
785 790 795 800
Gln Glu Leu Gln Pro Asp Val Pro Pro Ser Ser Gly Arg Ser
805 810
<210>326
<211>2848
<212>DNA
<213>Saccharomyces cerevisiae
<220>
<223>Kexin precursor(KEX2 protease) S08.048
<400>326
tgcataattc tgtcataagc ctgttctttt tcctggctta aacatcccgt tttgtaaaag 60
agaaatctat tccacatatt tcattcattc ggctaccata ctaaggataa actaatcccg 120
ttgttttttg gcctcgtcac ataattataa actactaacc cattatcaga tgaaagtgag 180
gaaatatatt actttatgct tttggtgggc cttttcaaca tccgctcttg tatcatcaca 240
acaaattcca ttgaaggacc atacgtcacg acagtatttt gctgtagaaa gcaatgaaac 300
attatcccgc ttggaggaaa tgcatccaaa ttggaaatat gaacatgatg ttcgagggct 360
accaaaccat tatgtttttt caaaagagtt gctaaaattg ggcaaaagat catcattaga 420
agagttacag ggggataaca acgaccacat attatctgtc catgatttat tcccgcgtaa 480
cgacctattt aagagactac cggtgcctgc tccaccaatg gactcaagct tgttaccggt 540
aaaagaagct gaggataaac tcagcataaa tgatccgctt tttgagaggc agtggcactt 600
ggtcaatcca agttttcctg gcagtgatat aaatgttctt gatctgtggt acaataatat 660
tacaggcgca ggggtcgtgg ctgccattgt tgatgatggc cttgactacg aaaatgaaga 720
cttgaaggat aatttttgcg ctgaaggttc ttgggatttc aacgacaata ccaatttacc 780
taaaccaaga ttatctgatg actaccatgg tacgagatgt gcaggtgaaa tagctgccaa 840
aaaaggtaac aatttttgcg gtgtcggggt aggttacaac gctaaaatct caggcataag 900
aatcttatcc ggtgatatca ctacggaaga tgaagctgcg tccttgattt atggtctaga 960
cgtaaacgat atatattcat gctcatgggg tcccgctgat gacggaagac atttacaagg 1020
ccctagtgac ctggtgaaaa aggctttagt aaaaggtgtt actgagggaa gagattccaa 1080
aggagcgatt tacgtttttg ccagtggaaa tggtggaact cgtggtgata attgcaatta 1140
cgacggctat actaattcca tatattctat tactattggg gctattgatc acaaagatct 1200
acatcctcct tattccgaag gttgttccgc cgtcatggca gtcacgtatt cttcaggttc 1260
aggcgaatat attcattcga gtgatatcaa cggcagatgc agtaatagcc acggtggaac 1320
gtctgcggct gctccattag ctgccggtgt ttacactttg ttactagaag ccaacccaaa 1380
cctaacttgg agagacgtac agtatttatc aatcttgtct gcggtagggt tagaaaagaa 1440
cgctgacgga gattggagag atagcgccat ggggaagaaa tactctcatc gctatggctt 1500
tggtaaaatc gatgcccata agttaattga aatgtccaag acctgggaga atgttaacgc 1560
acaaacctgg ttttacctgc caacattgta tgtttcccag tccacaaact ccacggaaga 1620
gacattagaa tccgtcataa ccatatcaga aaaaagtctt caagatgcta acttcaagag 1680
aattgagcac gtcacggtaa ctgtagatat tgatacagaa attaggggaa ctacgactgt 1740
cgatttaata tcaccagcgg ggataatttc aaaccttggc gttgtaagac caagagatgt 1800
ttcatcagag ggattcaaag actggacatt catgtctgta gcacattggg gtgagaacgg 1860
cgtaggtgat tggaaaatca aggttaagac aacagaaaat ggacacagga ttgacttcca 1920
cagttggagg ctgaagctct ttggggaatc cattgattca tctaaaacag aaactttcgt 1980
ctttggaaac gataaagagg aggttgaacc agctgctaca gaaagtaccg tatcacaata 2040
ttctgccagt tcaacttcta tttccatcag cgctacttct acatcttcta tctcaattgg 2100
tgtggaaacg tcggccattc cccaaacgac tactgcgagt accgatcctg attctgatcc 2160
aaacactcct aaaaaacttt cctctcctag gcaagccatg cattattttt taacaatatt 2220
tttgattggc gccacatttt tggtgttata cttcatgttt tttatgaaat caaggagaag 2280
gatcagaagg tcaagagcgg aaacgtatga attcgatatc attgatacag actctgagta 2340
cgattctact ttggacaatg gaacttccgg aattactgag cccgaagagg ttgaggactt 2400
cgattttgat ttgtccgatg aagaccatct tgcaagtttg tcttcatcag aaaacggtga 2460
tgctgaacat acaattgata gtgtactaac aaacgaaaat ccatttagtg accctataaa 2520
gcaaaagttc ccaaatgacg ccaacgcaga atctgcttcc aataaattac aagaattaca 2580
gcctgatgtt cctccatctt ccggacgatc gtgattcgat atgtacagaa agcttcaaat 2640
tacaaaatag catttttttc ttatagatta taatactctc tcatacgtat acgtatatgt 2700
gtatatgata tataaacaaa cattaatatc ctattccttc cgtttgaaat ccctatgatg 2760
tactttgcat tgtttgcacc cgcgaataaa atgaaaactc cgaaccgata tatcaagcac 2820
ataaaagggg agggtccaat taatgcat 2848
<210>327
<211>794
<212>PRT
<213>Homo sapiens
<220>
<223>Furin precursor S08.071
<400>327
Met Glu Leu Arg Pro Trp Leu Leu Trp Val Val Ala Ala Thr Gly Thr
1 5 10 15
Leu Val Leu Leu Ala Ala Asp Ala Gln Gly Gln Lys Val Phe Thr Asn
20 25 30
Thr Trp Ala Val Arg Ile Pro Gly Gly Pro Ala Val Ala Asn Ser Val
35 40 45
Ala Arg Lys His Gly Phe Leu Asn Leu Gly Gln Ile Phe Gly Asp Tyr
50 55 60
Tyr His Phe Trp His Arg Gly Val Thr Lys Arg Ser Leu Ser Pro His
65 70 75 80
Arg Pro Arg His Ser Arg Leu Gln Arg Glu Pro Gln Val Gln Trp Leu
85 90 95
Glu Gln Gln Val Ala Lys Arg Arg Thr Lys Arg Asp Val Tyr Gln Glu
100 105 110
Pro Thr Asp Pro Lys Phe Pro Gln Gln Trp Tyr Leu Ser Gly Val Thr
115 120 125
Gln Arg Asp Leu Asn Val Lys Ala Ala Trp Ala Gln Gly Tyr Thr Gly
130 135 140
His Gly Ile Val Val Ser Ile Leu Asp Asp Gly Ile Glu Lys Asn His
145 150 155 160
Pro Asp Leu Ala Gly Asn Tyr Asp Pro Gly Ala Ser Phe Asp Val Asn
165 170 175
Asp Gln Asp Pro Asp Pro Gln Pro Arg Tyr Thr Gln Met Asn Asp Asn
180 185 190
Arg His Gly Thr Arg Cys Ala Gly Glu Val Ala Ala Val Ala Asn Asn
195 200 205
Gly Val Cys Gly Val Gly Val Ala Tyr Asn Ala Arg Ile Gly Gly Val
210 215 220
Arg Met Leu Asp Gly Glu Val Thr Asp Ala Val Glu Ala Arg Ser Leu
225 230 235 240
Gly Leu Asn Pro Asn His Ile His Ile Tyr Ser Ala Ser Trp Gly Pro
245 250 255
Glu Asp Asp Gly Lys Thr Val Asp Gly Pro Ala Arg Leu Ala Glu Glu
260 265 270
Ala Phe Phe Arg Gly Val Ser Gln Gly Arg Gly Gly Leu Gly Ser Ile
275 280 285
Phe Val Trp Ala Ser Gly Asn Gly Gly Arg Glu His Asp Ser Cys Asn
290 295 300
Cys Asp Gly Tyr Thr Asn Ser Ile Tyr Thr Leu Ser Ile Ser Ser Ala
305 310 315 320
Thr Gln Phe Gly Asn Val Pro Trp Tyr Ser Glu Ala Cys Ser Ser Thr
325 330 335
Leu Ala Thr Thr Tyr Ser Ser Gly Asn Gln Asn Glu Lys Gln Ile Val
340 345 350
Thr Thr Asp Leu Arg Gln Lys Cys Thr Glu Ser His Thr Gly Thr Ser
355 360 365
Ala Ser Ala Pro Leu Ala Ala Gly Ile Ile Ala Leu Thr Leu Glu Ala
370 375 380
Asn Lys Asn Leu Thr Trp Arg Asp Met Gln His Leu Val Val Gln Thr
385 390 395 400
Ser Lys Pro Ala His Leu Asn Ala Asn Asp Trp Ala Thr Asn Gly Val
405 410 415
Gly Arg Lys Val Ser His Ser Tyr Gly Tyr Gly Leu Leu Asp Ala Gly
420 425 430
Ala Met Val Ala Leu Ala Gln Asn Trp Thr Thr Val Ala Pro Gln Arg
435 440 445
Lys Cys Ile Ile Asp Ile Leu Thr Glu Pro Lys Asp Ile Gly Lys Arg
450 455 460
Leu Glu Val Arg Lys Thr Val Thr Ala Cys Leu Gly Glu Pro Asn His
465 470 475 480
Ile Thr Arg Leu Glu His Ala Gln Ala Arg Leu Thr Leu Ser Tyr Asn
485 490 495
Arg Arg Gly Asp Leu Ala Ile His Leu Val Ser Pro Met Gly Thr Arg
500 505 510
Ser Thr Leu Leu Ala Ala Arg Pro His Asp Tyr Ser Ala Asp Gly Phe
515 520 525
Asn Asp Trp Ala Phe Met Thr Thr His Ser Trp Asp Glu Asp Pro Ser
530 535 540
Gly Glu Trp Val Leu Glu Ile Glu Asn Thr Ser Glu Ala Asn Asn Tyr
545 550 555 560
Gly Thr Leu Thr Lys Phe Thr Leu Val Leu Tyr Gly Thr Ala Pro Glu
565 570 575
Gly Leu Pro Val Pro Pro Glu Ser Ser Gly Cys Lys Thr Leu Thr Ser
580 585 590
Ser Gln Ala Cys Val Val Cys Glu Glu Gly Phe Ser Leu His Gln Lys
595 600 605
Ser Cys Val Gln His Cys Pro Pro Gly Phe Ala Pro Gln Val Leu Asp
610 615 620
Thr His Tyr Ser Thr Glu Asn Asp Val Glu Thr Ile Arg Ala Ser Val
625 630 635 640
Cys Ala Pro Cys His Ala Ser Cys Ala Thr Cys Gln Gly Pro Ala Leu
645 650 655
Thr Asp Cys Leu Ser Cys Pro Ser His Ala Ser Leu Asp Pro Val Glu
660 665 670
Gln Thr Cys Ser Arg Gln Ser Gln Ser Ser Arg Glu Ser Pro Pro Gln
675 680 685
Gln Gln Pro Pro Arg Leu Pro Pro Glu Val Glu Ala Gly Gln Arg Leu
690 695 700
Arg Ala Gly Leu Leu Pro Ser His Leu Pro Glu Val Val Ala Gly Leu
705 710 715 720
Ser Cys Ala Phe Ile Val Leu Val Phe Val Thr Val Phe Leu Val Leu
725 730 735
Gln Leu Arg Ser Gly Phe Ser Phe Arg Gly Val Lys Val Tyr Thr Met
740 745 750
Asp Arg Gly Leu Ile Ser Tyr Lys Gly Leu Pro Pro Glu Ala Trp Gln
755 760 765
Glu Glu Cys Pro Ser Asp Ser Glu Glu Asp Glu Gly Arg Gly Glu Arg
770 775 780
Thr Ala Phe Ile Lys Asp Gln Ser Ala Leu
785 790
<210>328
<211>4180
<212>DNA
<213>Homo sapiens
<220>
<223>Furin precursor S08.071
<400>328
gcggggaagc agcagcggcc aggatgaatc ccaggtgctc tggagctgga tggtgaaggt 60
cggcactctt caccctcccg agccctgccc gtctcggccc catgccccca ccagtcagcc 120
ccgggccaca ggcagtgagc aggcacctgg gagccgaggc cctatgacca ggccaaggag 180
acgggcgctc cagggtccca gccacctgtc ccccccatgg agctgaggcc ctggttgcta 240
tgggtggtag cagcaacagg aaccttggtc ctgctagcag ctgatgctca gggccagaag 300
gtcttcacca acacgtgggc tgtgcgcatc cctggaggcc cagcggtggc caacagtgtg 360
gcacggaagc atgggttcct caacctgggc cagatcttcg gggactatta ccacttctgg 420
catcgaggag tgacgaagcg gtccctgtcg cctcaccgcc cgcggcacag ccggctgcag 480
agggagcctc aagtacagtg gctggaacag caggtggcaa agcgacggac taaacgggac 540
gtgtaccagg agcccacaga ccccaagttt cctcagcagt ggtacctgtc tggtgtcact 600
cagcgggacc tgaatgtgaa ggcggcctgg gcgcagggct acacagggca cggcattgtg 660
gtctccattc tggacgatgg catcgagaag aaccacccgg acttggcagg caattatgat 720
cctggggcca gttttgatgt caatgaccag gaccctgacc cccagcctcg gtacacacag 780
atgaatgaca acaggcacgg cacacggtgt gcgggggaag tggctgcggt ggccaacaac 840
ggtgtctgtg gtgtaggtgt ggcctacaac gcccgcattg gaggggtgcg catgctggat 900
ggcgaggtga cagatgcagt ggaggcacgc tcgctgggcc tgaaccccaa ccacatccac 960
atctacagtg ccagctgggg ccccgaggat gacggcaaga cagtggatgg gccagcccgc 1020
ctcgccgagg aggccttctt ccgtggggtt agccagggcc gaggggggct gggctccatc 1080
tttgtctggg cctcggggaa cgggggccgg gaacatgaca gctgcaactg cgacggctac 1140
accaacagta tctacacgct gtccatcagc agcgccacgc agtttggcaa cgtgccgtgg 1200
tacagcgagg cctgctcgtc cacactggcc acgacctaca gcagtggcaa ccagaatgag 1260
aagcagatcg tgacgactga cttgcggcag aagtgcacgg agtctcacac gggcacctca 1320
gcctctgccc ccttagcagc cggcatcatt gctctcaccc tggaggccaa taagaacctc 1380
acatggcggg acatgcaaca cctggtggta cagacctcga agccagccca cctcaatgcc 1440
aacgactggg ccaccaatgg tgtgggccgg aaagtgagcc actcatatgg ctacgggctt 1500
ttggacgcag gcgccatggt ggccctggcc cagaattgga ccacagtggc cccccagcgg 1560
aagtgcatca tcgacatcct caccgagccc aaagacatcg ggaaacggct cgaggtgcgg 1620
aagaccgtga ccgcgtgcct gggcgagccc aaccacatca ctcggctgga gcacgctcag 1680
gcgcggctca ccctgtccta taatcgccgt ggcgacctgg ccatccacct ggtcagcccc 1740
atgggcaccc gctccaccct gctggcagcc aggccacatg actactccgc agatgggttt 1800
aatgactggg ccttcatgac aactcattcc tgggatgagg atccctctgg cgagtgggtc 1860
ctagagattg aaaacaccag cgaagccaac aactatggga cgctgaccaa gttcaccctc 1920
gtactctatg gcaccgcccc tgaggggctg cccgtacctc cagaaagcag tggctgcaag 1980
accctcacgt ccagtcaggc ctgtgtggtg tgcgaggaag gcttctccct gcaccagaag 2040
agctgtgtcc agcactgccc tccaggcttc gccccccaag tcctcgatac gcactatagc 2100
accgagaatg acgtggagac catccgggcc agcgtctgcg ccccctgcca cgcctcatgt 2160
gccacatgcc aggggccggc cctgacagac tgcctcagct gccccagcca cgcctccttg 2220
gaccctgtgg agcagacttg ctcccggcaa agccagagca gccgagagtc cccgccacag 2280
cagcagccac ctcggctgcc cccggaggtg gaggcggggc aacggctgcg ggcagggctg 2340
ctgccctcac acctgcctga ggtggtggcc ggcctcagct gcgccttcat cgtgctggtc 2400
ttcgtcactg tcttcctggt cctgcagctg cgctctggct ttagttttcg gggggtgaag 2460
gtgtacacca tggaccgtgg cctcatctcc tacaaggggc tgccccctga agcctggcag 2520
gaggagtgcc cgtctgactc agaagaggac gagggccggg gcgagaggac cgcctttatc 2580
aaagaccaga gcgccctctg atgagcccac tgcccacccc ctcaagccaa tcccctcctt 2640
gggcactttt taattcacca aagtattttt ttatcttggg actgggtttg gaccccagct 2700
gggaggcaag aggggtggag actgtttccc atcctaccct cgggcccacc tggccacctg 2760
aggtgggccc aggaccagct ggggcgtggg gagggccgta ccccaccctc agcacccctt 2820
ccatgtggag aaaggagtga aacctttagg gcagcttgcc ccggccccgg ccccagccag 2880
agttcctgcg gagtgaagag gggcagccct tgcttgttgg gattcctgac ccaggccgca 2940
gctcttgccc ttccctgtcc ctctaaagca ataatggtcc catccaggca gtcgggggct 3000
ggcctaggag atatctgagg gaggaggcca cctctccaag ggcttctgca ccctccaccc 3060
tgtcccccag ctctggtgag tcttggcggc agcagccatc ataggaaggg accaaggcaa 3120
ggcaggtgcc tccaggtgtg cacgtggcat gtggcctgtg gcctgtgtcc catgacccac 3180
ccctgtgctc cgtgcctcca ccaccactgg ccaccaggct ggcgcagcca aggccgaagc 3240
tctggctgaa ccctgtgctg gtgtcctgac caccctcccc tctcttgcac ccgcctctcc 3300
cgtcagggcc caagtccctg ttttctgagc ccgggctgcc tgggctgttg gcactcacag 3360
acctggagcc cctgggtggg tggtggggag gggcgctggc ccagccggcc tctctggcct 3420
cccacccgat gctgctttcc cctgtgggga tctcaggggc tgtttgagga tatattttca 3480
ctttgtgatt atttcacttt agatgctgat gatttgtttt tgtattttta atgggggtag 3540
cagctggact acccacgttc tcacacccac cgtccgccct gctcctccct ggctgccctg 3600
gccctgaggt gtgggggctg cagcatgttg ctgaggagtg aggaatagtt gagccccaag 3660
tcctgaagag gcgggccagc caggcgggct caaggaaagg gggtcccagt gggaggggca 3720
ggctgacatc tgtgtttcaa gtggggctcg ccatgccggg ggttcatagg tcactggctc 3780
tccaagtgcc agaggtgggc aggtggtggc actgagcccc cccaacactg tgccctggtg 3840
gagaaagcac tgacctgtca tgcccccctc aaacctcctc ttctgacgtg ccttttgcac 3900
ccctcccatt aggacaatca gtcccctccc atctgggagt ccccttttct tttctaccct 3960
agccattcct ggtacccagc catctgccca ggggtgcccc ctcctctccc atccccctgc 4020
cctcgtggcc agcccggctg gttttgtaag atactgggtt ggtgcacagt gatttttttc 4080
ttgtaattta aacaggccca gcattgctgg ttctatttaa tggacatgag ataatgttag 4140
aggttttaaa gtgattaaac gtgcagacta tgcaaaccag 4180
<210>329
<211>969
<212>PRT
<213>Homo sapiens
<220>
<223>Subtilisin/kexin-like protease PACE4 S08.075
<400>329
Met Pro Pro Arg Ala Pro Pro Ala Pro Gly Pro Arg Pro Pro Pro Arg
1 5 10 15
Ala Ala Ala Ala Thr Asp Thr Ala Ala Gly Ala Gly Gly Ala Gly Gly
20 25 30
Ala Gly Gly Ala Gly Gly Pro Gly Phe Arg Pro Leu Ala Pro Arg Pro
35 40 45
Trp Arg Trp Leu Leu Leu Leu Ala Leu Pro Ala Ala Cys Ser Ala Pro
50 55 60
Pro Pro Arg Pro Val Tyr Thr Asn His Trp Ala Val Gln Val Leu Gly
65 70 75 80
Gly Pro Ala Glu Ala Asp Arg Val Ala Ala Ala His Gly Tyr Leu Asn
85 90 95
Leu Gly Gln Ile Gly Asn Leu Glu Asp Tyr Tyr His Phe Tyr His Ser
100 105 110
Lys Thr Phe Lys Arg Ser Thr Leu Ser Ser Arg Gly Pro His Thr Phe
115 120 125
Leu Arg Met Asp Pro Gln Val Lys Trp Leu Gln Gln Gln G1u Val Lys
130 135 140
Arg Arg Val Lys Arg Gln Val Arg Ser Asp Pro Gln Ala Leu Tyr Phe
145 150 155 160
Asn Asp Pro Ile Trp Ser Asn Met Trp Tyr Leu His Cys Gly Asp Lys
165 170 175
Asn Ser Arg Cys Arg Ser Glu Met Asn Val Gln Ala Ala Trp Lys Arg
180 185 190
Gly Tyr Thr Gly Lys Asn Val Val Val Thr Ile Leu Asp Asp Gly Ile
195 200 205
Glu Arg Asn His Pro Asp Leu Ala Pro Asn Tyr Asp Ser Tyr Ala Ser
210 215 220
Tyr Asp Val Asn Gly Asn Asp Tyr Asp Pro Ser Pro Arg Tyr Asp Ala
225 230 235 240
Ser Asn Glu Asn Lys His Gly Thr Arg Cys Ala Gly Glu Val Ala Ala
245 250 255
Ser Ala Asn Asn Ser Tyr Cys Ile Val Gly Ile Ala Tyr Asn Ala Lys
260 265 270
Ile Gly Gly Ile Arg Met Leu Asp Gly Asp Val Thr Asp Val Val Glu
275 280 285
Ala Lys Ser Leu Gly Ile Arg Pro Asn Tyr Ile Asp Ile Tyr Ser Ala
290 295 300
Ser Trp Gly Pro Asp Asp Asp Gly Lys Thr Val Asp Gly Pro Gly Arg
305 310 315 320
Leu Ala Lys Gln Ala Phe Glu Tyr Gly Ile Lys Lys Gly Arg Gln Gly
325 330 335
Leu Gly Ser Ile Phe Val Trp Ala Ser Gly Asn Gly Gly Arg Glu Gly
340 345 350
Asp Tyr Cys Ser Cys Asp Gly Tyr Thr Asn Ser Ile Tyr Thr Ile Ser
355 360 365
Val Ser Ser Ala Thr Glu Asn Gly Tyr Lys Pro Trp Tyr Leu Glu Glu
370 375 380
Cys Ala Ser Thr Leu Ala Thr Thr Tyr Ser Ser Gly Ala Phe Tyr Glu
385 390 395 400
Arg Lys Ile Val Thr Thr Asp Leu Arg Gln Arg Cys Thr Asp Gly His
405 410 415
Thr Gly Thr Ser Val Ser Ala Pro Met Val Ala Gly Ile Ile Ala Leu
420 425 430
Ala Leu Glu Ala Asn Ser Gln Leu Thr Trp Arg Asp Val Gln His Leu
435 440 445
Leu Val Lys Thr Ser Arg Pro Ala His Leu Lys Ala Ser Asp Trp Lys
450 455 460
Val Asn Gly Ala Gly His Lys Val Ser His Phe Tyr Gly Phe Gly Leu
465 470 475 480
Val Asp Ala Glu Ala Leu Val Val Glu Ala Lys Lys Trp Thr Ala Val
485 490 495
Pro Ser Gln His Met Cys Val Ala Ala Ser Asp Lys Arg Pro Arg Ser
500 505 510
Ile Pro Leu Val Gln Val Leu Arg Thr Thr Ala Leu Thr Ser Ala Cys
515 520 525
Ala Glu His Ser Asp Gln Arg Val Val Tyr Leu Glu His Val Val Val
530 535 540
Arg Thr Ser Ile Ser His Pro Arg Arg Gly Asp Leu Gln Ile Tyr Leu
545 550 555 560
Val Ser Pro Ser Gly Thr Lys Ser Gln Leu Leu Ala Lys Arg Leu Leu
565 570 575
Asp Leu Ser Asn Glu Gly Phe Thr Asn Trp Glu Phe Mer Thr Val His
580 585 590
Cys Trp Gly Glu Lys Ala Glu Gly Gln Trp Thr Leu Glu Ile Gln Asp
595 600 605
Leu Pro Ser Gln Val Arg Asn Pro Glu Lys Gln Gly Lys Leu Lys Glu
610 615 620
Trp Ser Leu Ile Leu Tyr Gly Thr Ala Glu His Pro Tyr His Thr Phe
625 630 635 640
Ser Ala His Gln Ser Arg Ser Arg Met Leu Glu Leu Ser Ala Pro Glu
645 650 655
Leu Glu Pro Pro Lys Ala Ala Leu Ser Pro Ser Gln Val Glu Val Pro
660 665 670
Glu Asp Glu Glu Asp Tyr Thr Ala Gln Ser Thr Pro Gly Ser Ala Asn
675 680 685
Ile Leu Gln Thr Ser Val Cys His Pro Glu Cys Gly Asp Lys Gly Cys
690 695 700
Asp Gly Pro Asn Ala Asp Gln Cys Leu Asn Cys Val His Phe Ser Leu
705 710 715 720
Gly Ser Val Lys Thr Ser Arg Lys Cys Val Ser Val Cys Pro Leu Gly
725 730 735
Tyr Phe Gly Asp Thr Ala Ala Arg Arg Cys Arg Arg Cys His Lys Gly
740 745 750
Cys Glu Thr Cys Ser Ser Arg Ala Ala Thr Gln Cys Leu Ser Cys Arg
755 760 765
Arg Gly Phe Tyr His His Gln Glu Met Asn Thr Cys Val Thr Leu Cys
770 775 780
Pro Ala Gly Phe Tyr Ala Asp Glu Ser Gln Lys Asn Cys Leu Lys Cys
785 790 795 800
His Pro Ser Cys Lys Lys Cys Val Asp Glu Pro Glu Lys Cys Thr Val
805 810 815
Cys Lys Glu Gly Phe Ser Leu Ala Arg Gly Ser Cys Ile Pro Asp Cys
820 825 830
Glu Pro Gly Thr Tyr Phe Asp Ser Glu Leu Ile Arg Cys Gly Glu Cys
835 840 845
His His Thr Cys Gly Thr Cys Val Gly Pro Gly Arg Glu Glu Cys Ile
850 855 860
His Cys Ala Lys Asn Phe His Phe His Asp Trp Lys Cys Val Pro Ala
865 870 875 880
Cys Gly Glu Gly Phe Tyr Pro Glu Glu Met Pro Gly Leu Pro His Lys
885 890 895
Val Cys Arg Arg Cys Asp Glu Asn Cys Leu Ser Cys Ala Gly Ser Ser
900 905 910
Arg Asn Cys Ser Arg Cys Lys Thr Gly Phe Thr Gln Leu Gly Thr Ser
915 920 925
Cys Ile Thr Asn His Thr Cys Ser Asn Ala Asp Glu Thr Phe Cys Glu
930 935 940
Met Val Lys Ser Asn Arg Leu Cys Glu Arg Lys Leu Phe Ile Gln Phe
945 950 955 960
Cys Cys Arg Thr Cys Leu Leu Ala Gly
965
<210>330
<211>4403
<212>DNA
<213>Homo sapiens
<220>
<223>Subtilisin/kexin-like protease PACE4 S08.075
<400>330
cgggaacgcg ccgcggccgc ctcctcctcc ccggctcccg cccgcggcgg tgttggcggc 60
ggcggtggcg gcggcggcgg cgcttccccg gcgcggagcg gctttaaaag gcggcactcc 120
accccccggc gcactcgcag ctcgggcgcc gcgcgagcct gtcgccgcta tgcctccgcg 180
cgcgccgcct gcgcccgggc cccggccgcc gccccgggcc gccgccgcca ccgacaccgc 240
cgcgggcgcg gggggcgcgg ggggcgcggg gggcgccggc gggcccgggt tccggccgct 300
cgcgccgcgt ccctggcgct ggctgctgct gctggcgctg cctgccgcct gctccgcgcc 360
cccgccgcgc cccgtctaca ccaaccactg ggcggtgcaa gtgctgggcg gcccggccga 420
ggcggaccgc gtggcggcgg cgcacggcta cctcaacttg ggccagattg gaaacctgga 480
agattactac catttttatc acagcaaaac ctttaaaaga tcaaccttga gtagcagagg 540
ccctcacacc ttcctcagaa tggaccccca ggtgaaatgg ctccagcaac aggaagtgaa 600
acgaagggtg aagagacagg tgcgaagtga cccgcaggcc ctttacttca acgaccccat 660
ttggtccaac atgtggtacc tgcattgtgg cgacaagaac agtcgctgcc ggtcggaaat 720
gaatgtccag gcagcgtgga agaggggcta cacaggaaaa aacgtggtgg tcaccatcct 780
tgatgatggc atagagagaa atcaccctga cctggcccca aattatgatt cctacgccag 840
ctacgacgtg aacggcaatg attatgaccc atctccacga tatgatgcca gcaatgaaaa 900
taaacacggc actcgttgtg cgggagaagt tgctgcttca gcaaacaatt cctactgcat 960
cgtgggcata gcgtacaatg ccaaaatagg aggcatccgc atgctggacg gcgatgtcac 1020
agatgtggtc gaggcaaagt cgctgggcat cagacccaac tacatcgaca tttacagtgc 1080
cagctggggg ccggacgacg acggcaagac ggtggacggg cccggccgac tggctaagca 1140
ggctttcgag tatggcatta aaaagggccg gcagggcctg ggctccattt tcgtctgggc 1200
atctgggaat ggcgggagag agggggacta ctgctcgtgc gatggctaca ccaacagcat 1260
ctacaccatc tccgtcagca gcgccaccga gaatggctac aagccctggt acctggaaga 1320
gtgtgcctcc accctggcca ccacctacag cagtggggcc ttttatgagc gaaaaatcgt 1380
caccacggat ctgcgtcagc gctgtaccga tggccacact gggacctcag tctctgcccc 1440
catggtggcg ggcatcatcg ccttggctct agaagcaaac agccagttaa cctggaggga 1500
cgtccagcac ctgctagtga agacatcccg gccggcccac ctgaaagcga gcgactggaa 1560
agtaaacggc gcgggtcata aagttagcca tttctatgga tttggtttgg tggacgcaga 1620
agctctcgtt gtggaggcaa agaagtggac agcagtgcca tcgcagcaca tgtgtgtggc 1680
cgcctcggac aagagaccca ggagcatccc cttagtgcag gtgctgcgga ctacggccct 1740
gaccagcgcc tgcgcggagc actcggacca gcgggtggtc tacttggagc acgtggtggt 1800
tcgcacctcc atctcacacc cacgccgagg agacctccag atctacctgg tttctccctc 1860
gggaaccaag tctcaacttt tggcaaagag gttgctggat ctttccaatg aagggtttac 1920
aaactgggaa ttcatgactg tccactgctg gggagaaaag gctgaagggc agtggacctt 1980
ggaaatccaa gatctgccat cccaggtccg caacccggag aagcaaggga agttgaaaga 2040
atggagcctc atactgtatg gcacagcaga gcacccgtac cacaccttca gtgcccatca 2100
gtcccgctcg cggatgctgg agctctcagc cccagagctg gagccaccca aggctgccct 2160
gtcaccctcc caggtggaag ttcctgaaga tgaggaagat tacacagctc aatccacccc 2220
aggctctgct aatattttac agaccagtgt gtgccatccg gagtgtggtg acaaaggctg 2280
tgatggcccc aatgcagacc agtgcttgaa ctgcgtccac ttcagcctgg ggagtgtcaa 2340
gaccagcagg aagtgcgtga gtgtgtgccc cttgggctac tttggggaca cagcagcaag 2400
acgctgtcgc cggtgccaca aggggtgtga gacctgctcc agcagagctg cgacgcagtg 2460
cctgtcttgc cgccgcgggt tctatcacca ccaggagatg aacacctgtg tgaccctctg 2520
tcctgcagga ttttatgctg atgaaagtca gaaaaattgc cttaaatgcc acccaagctg 2580
taaaaagtgc gtggatgaac ctgagaaatg tactgtctgt aaagaaggat tcagccttgc 2640
acggggcagc tgcattcctg actgtgagcc aggcacctac tttgactcag agctgatcag 2700
atgtggggaa tgccatcaca cctgcggaac ctgcgtgggg ccaggcagag aagagtgcat 2760
tcactgtgcg aaaaacttcc acttccacga ctggaagtgt gtgccagcct gtggtgaggg 2820
cttctaccca gaagagatgc cgggcttgcc ccacaaagtg tgtcgaaggt gtgacgagaa 2880
ctgcttgagc tgtgcaggct ccagcaggaa ctgtagcagg tgtaagacgg gcttcacaca 2940
gctggggacc tcctgcatca ccaaccacac gtgcagcaac gctgacgaga cattctgcga 3000
gatggtgaag tccaaccggc tgtgcgaacg gaagctcttc attcagttct gctgccgcac 3060
gtgcctcctg gccgggtaag ggtgcctagc tgcccacaga gggcaggcac tcccatccat 3120
ccatccgtcc accttcctcc agactgtcgg ccagagtctg tttcaggagc ggcgccctgc 3180
acctgacagc tttatctccc caggagcagc atctctgagc acccaagcca ggtgggtggt 3240
ggctcttaag gaggtgttcc taaaatggtg atatcctctc aaatgctgct tgttggctcc 3300
agtcttccga caaactaaca ggaacaaaat gaattctggg aatccacagc tctggctttg 3360
gagcagcttc tgggaccata agtttactga atcttcaaga ccaaagcaga aaagaaaggc 3420
gcttggcatc acacatcact cttctccccg tgcttttctg cggctgtgta gtaaatctcc 3480
ccggcccagc tggcgaaccc tgggccatcc tcacatgtga caaagggcca gcagtctacc 3540
tgctcgttgc ctgccactga gcagtctggg gacggtttgg tcagactata aataagatag 3600
gtttgagggc ataaaatgta tgaccactgg ggccggagta tctatttcta catagtcagc 3660
tacttctgaa actgcagcag tggcttagaa agtccaattc caaagccaga ccagaagatt 3720
ctatcccccg cagcgctctc ctttgagcaa gccgagctct ccttgttacc gtgttctgtc 3780
tgtgtcttca ggagtctcat ggcctgaacg accacctcga cctgatgcag agccttctga 3840
ggagaggcaa caggaggcat tctgtggcca gccaaaaggt accccgatgg ccaagcaatt 3900
cctctgaaca aaatgtaaag ccagccatgc attgttaatc atccatcact tcccatttta 3960
tggaattgct tttaaaatac atttggcctc tgcccttcag aagactcgtt tttaaggtgg 4020
aaactcctgt gtctgtgtat attacaagcc tacatgacac agttggattt attctgccaa 4080
acctgtgtag gcattttata agctacatgt tctaattttt accgatgtta attattttga 4140
caaatatttc atatattttc attgaaatgc acagatctgc ttgatcaatt cccttgaata 4200
gggaagtaac atttgcctta aattttttcg acctcgtctt tctccatatt gtcctgctcc 4260
cctgtttgac gacagtgcat ttgccttgtc acctgtgagc tggagagaac ccagatgttg 4320
tttattgaat ctacaactct gaaagagaaa tcaatgaagc aagtacaatg ttaaccctaa 4380
attaataaaa gagttaacat ccc 4403
<210>331
<211>662
<212>PRT
<213>Anabaena variabilis
<220>
<223>Calcium-dependent protease precursor(Trypsin)
S08.79
<400>331
Met Val His Val Arg Tyr Gly Gly Gln Asn Gly Glu Gln Tyr Glu Leu
1 5 10 15
Ala Ile Ser Glu Asn His Ile Val Val Arg Thr Glu Ser Arg Ser Ser
20 25 30
Leu Ile Ser Asp Arg Pro Phe Glu Ala Ala Pro Val Ser Pro Gln Ala
35 40 45
Arg Asn Ile Leu Asn Gln Phe Glu Leu Ser Thr Arg Phe Ser Gln Ala
50 55 60
Gly Val Glu Val Leu His Val Lys Glu Pro Ser His Asp Gly Ala Leu
65 70 75 80
Arg Asp Thr Ala Arg Glu Ile Leu Asn Gln Glu Pro Glu Val Gln Phe
85 90 95
Ala Gly Arg Val Leu Ile Asp Pro Val Ser Gln Gln Pro Ile Val Tyr
100 105 110
Thr Glu Asn Leu Phe Val Lys Phe Asp His Glu Glu Asp Val Ser Phe
115 120 125
Cys Gln Glu Ile Leu Gly Arg Tyr Gly Leu Thr Ile Lys Arg Gln Leu
130 135 140
Glu Tyr Ala Arg Asn Ala Tyr Phe Val Ser Ala Pro Ser Asn Thr Gly
145 150 155 160
Leu Ala Ile Phe Asp Ile Ser Glu Arg Leu Leu Asn Glu Glu Ser Val
165 170 175
Glu Leu Cys His Pro Glu Leu Val Arg Glu Phe Arg Gln Arg Gln Ala
180 185 190
Phe Pro Pro Gln Trp His Leu Lys Gln Thr Thr Ile Gly Gly Lys Thr
195 200 205
Ile Asn Ala His Ala Asn Val Glu Ala Ala Trp Lys Leu Ser Asp Gly
210 215 220
Thr Gly Thr Ile Ile Ala Ile Ile Asp Asp Gly Val Asp Ile Asp His
225 230 235 240
Glu Glu Phe Arg Ser Ser Gly Lys Ile Val Ala Pro Arg Asp Val Thr
245 250 255
Arg Lys Thr Asn Phe Pro Thr Pro Gly Asn Arg Asp Asn His G1y Thr
260 265 270
Ala Cys Ala Gly Val Ala Cys Gly Asn Gly Asn Phe Gly Ala Ser Gly
275 280 285
Val Ala Pro Gly Ala Lys Leu Met Pro Ile Arg Phe Val Ser Ala Leu
290 295 300
Gly Ser Gln Asp Glu Ala Asp Ser Phe Val Trp Ala Ala Gln Asn Gly
305 310 315 320
Ala Asp Val Ile Ser Cys Ser Trp Gly Pro Pro Asp Gly Thr Trp Trp
325 330 335
Asp Asp Lys Asp Pro Leu His Lys Gln Lys Val Pro Leu Pro Asp Ser
340 345 350
Thr Arg Leu Ala Met Asp Tyr Ala Ile Asn Lys Gly Arg Asn Gly Lys
355 360 365
Gly Cys Val Ile Leu Phe Ala Ala Gly Asn Gly Asn Glu Ser Val Asp
370 375 380
Asn Asp Gly Tyr Ala Ser Tyr Glu Lys Val Ile Ala Val Ala Ala Cys
385 390 395 400
Asn Asp Phe Gly Thr Arg Ser Ala Tyr Ser Asp Phe Gly Thr Ala Val
405 410 415
Trp Cys Ala Phe Pro Ser Asn Asn Gly Asn Pro Ser Gln Thr Pro Gly
420 425 430
Ile Trp Thr Ala Asp Arg Thr Gly Val Val Gly Tyr Asn Ser Gly Asn
435 440 445
Thr Asn Leu Gly Asp Gln Ala Gly Asn Tyr Thr Asn Ser Phe Gly Gly
450 455 460
Thr Ser Ser Ala Cys Pro Gly Ala Ala Gly Val Ala Ala Leu Ile Leu
465 470 475 480
Ser Arg Asn Pro Asn Leu Arg Trp Asp Glu Val Arg Asp Ile Ile Lys
485 490 495
Arg Ser Cys Asp Arg Ile Asp Pro Val Gly Gly Asn Tyr Asn Ala Glu
500 505 510
Gly Arg Ser Pro Phe Tyr Gly Tyr Gly Arg Ile Asn Ala Leu Lys Ala
515 520 525
Val Glu Leu Ala Leu Pro Ala Gln Pro Glu Pro Val Ser Ile Phe Thr
530 535 540
Ala Val Gln Asp Val Pro Ile Asn Asp Leu Gln Ile Ser Gln Leu Ser
545 550 555 560
Leu Ala Ile Ala Asn Thr Asn Pro Ile Lys Ser Ile Lys Val Thr Val
565 570 575
Asp Ile Glu His Thr Tyr Ile Gly Asp Leu Val Val Ser Leu Asn Pro
580 585 590
Pro Ala Glu Ser Gly Val Leu Pro Ile Ile Leu His Asp Arg Lys Gly
595 600 605
Gly Gly Ala Asp Asp Ile Lys Gln Thr Tyr Asp Glu Val Ser Thr Pro
610 615 620
Gly Leu Thr Ala Leu Lys Gly Lys Ile Pro Gln Gly Thr Trp Thr Leu
625 630 635 640
Glu Val Ala Asp Lys Ala Gln Ala Asp Thr Gly Lys Ile Arg Ser Leu
645 650 655
Thr Ile Glu Leu Gly Phe
660
<210>332
<211>2187
<212>DNA
<213>Anabaena variabilis
<220>
<223>Calcium-dependent protease precursor
(Trypsin)S08.79
<400>332
ggtaggctgt tgattttgtg ccttttcagg gttgaaaaat ttatgcacta tcctgttcta 60
gctgagtaaa atttgcagtg ggaaaagttc attacccatt acccattacc catttctgga 120
aagccagact gatagtttgt atccacagga aattgttcta agctgcgtct tatgtgtgcg 180
tcattttgga aacagcacac aaaatacaga ggcttagtat caacttaata gtaggagatt 240
aagtatggtt catgtccgct atggtggtca gaacggtgaa cagtacgaat tagcgattag 300
tgagaatcat attgtagtac ggactgaaag ccgcagctcg ctaattagcg atcgcccctt 360
tgaagccgcc cctgtttcac cccaagcccg taatatcctc aatcaattcg agttatcaac 420
tcgtttcagc caagcaggtg tagaagttct ccatgtgaag gaaccaagcc acgatggtgc 480
tttgcgcgac acagccaggg aaattctcaa tcaagaacct gaggtgcaat ttgccggacg 540
tgtgttaatt gatccggtca gtcaacaacc catagtttac acagaaaacc tcttcgtgaa 600
gtttgaccac gaagaagacg ttagtttctg ccaagaaatt ttgggacgct acggcttaac 660
aattaaacgt caactagaat acgcccgtaa tgcctacttt gtcagcgcac cctccaatac 720
tggtttagcc atctttgata tttccgaaag gttgctcaac gaggaatcgg tggaattgtg 780
tcatcctgaa ttggtgcggg aatttagaca acgtcaggca tttcccccgc aatggcatct 840
caagcaaacc acgattgggg gtaaaactat caatgcccac gccaatgttg aggcggcatg 900
gaagttgagc gatggtacgg gaacgattat tgccattatt gatgatggtg tggatatcga 960
ccatgaagag ttccgttcct ctggtaagat tgtcgcccct agagatgtca cacggaaaac 1020
caactttccc acaccaggaa accgagataa tcacggtacg gcttgtgcag gtgtagcttg 1080
tggtaatggc aactttgggg cttctggggt agcgcctggg gctaagttaa tgcccattcg 1140
tttcgtttct gccctgggtt cccaagatga agccgactct tttgtctggg ctgctcaaaa 1200
tggtgcggat gtaatttcct gtagttgggg gccaccagat ggtacatggt gggatgataa 1260
agacccccta cacaagcaga aagtacctct acccgattcc acaagactag caatggacta 1320
tgccatcaat aaggggcgta atggcaaagg atgtgtgatt ttattcgctg cgggtaatgg 1380
taacgaaagc gtagataatg acggctacgc cagctatgaa aaggtcatag ctgtcgcggc 1440
ttgtaacgac tttggtacta ggagtgctta cagtgatttt ggtacagccg tctggtgtgc 1500
ctttcctagc aacaacggta acccttccca aacccctggt atttggacag ccgatcgcac 1560
tggtgtagtc ggttataatt ccggtaatac taatctcggt gaccaggcag gcaattatac 1620
aaatagtttc ggtggcacct ctagcgcctg tcctggtgca gctggtgtcg ccgccttgat 1680
tctctctcgc aatccaaatc tgcgttggga tgaggtcaga gatattatta aacgctcatg 1740
cgatcgcatt gatccagttg gtggtaacta taacgctgaa ggtcgtagcc cattctacgg 1800
ttatggtcgt atcaatgccc tcaaagcggt agaattggca ctacccgcac aacctgagcc 1860
agtcagcatt ttcacggctg tgcaggatgt ccccatcaac gacttgcaga tatctcaact 1920
cagtttagcg atcgctaaca ccaaccccat caaatctatc aaagttacgg tagatatcga 1980
gcatacctac atcggtgact tagtagttag cctcaacccc cctgctgaat ccggtgtatt 2040
accaatcatc ctccacgacc gcaaaggcga ggcgcagatg atatcaaaca aacttacgat 2100
gaggtgagta caccaggact aactgctttg aagggtaaaa ttcctcaagg aacctggact 2160
ttagaagtag cagataaagc acaggcg 2187
<210>333
<211>402
<212>PRT
<213>Rattus norvegicus
<220>
<223>Caspase-1 precursor C14.001
<400>333
Met Ala Asp Lys Val Leu Arg Ala Lys Arg Lys Gln Phe Ile Asn Ser
1 5 10 15
Val Ser Val Gly Thr Ile Asn Gly Leu Leu Asp Glu Leu Leu Glu Lys
20 25 30
Arg Val Leu Asn Gln Glu Glu Met Asp Thr Ile Lys Leu Ala Asn Ile
35 40 45
Thr Val Met Glu Lys Ala Arg Asp Leu Cys Asp His Val Thr Lys Lys
50 55 60
Gly Pro Arg Ala Ser Gln Met Phe Ile Thr Tyr Ile Cys Asn Glu Asp
65 70 75 80
Cys Tyr Leu Ala Glu Ile Leu Glu Leu Gln Ser Gly Pro Ser Ala Glu
85 90 95
Thr Val Phe Val Thr Glu Asp Ser Lys Gly Gly His Pro Phe Ser Ser
100 105 110
Glu Thr Lys Glu Lys Leu Asn Lys Glu Gly Gly Ala Phe Pro Gly Pro
115 120 125
Ser Gly Ser Leu Lys Phe Cys Pro Leu Glu Ile Ala Gln Lys Leu Trp
130 135 140
Lys Glu Asn His Ser Glu Ile Tyr Pro Ile Met Lys Thr Pro Thr Arg
145 150 155 160
Thr Arg Leu Ala Leu Ile Ile Cys Asn Thr Asp Phe Gln His Leu Ser
165 170 175
Arg Arg Val Gly Ala Asp Val Asp Leu Arg Glu Met Lys Leu Leu Leu
180 185 190
Gln Asp Leu Gly Tyr Thr Val Lys Val Lys Glu Asn Leu Thr Ala Leu
195 200 205
Glu Met Thr Lys Glu Leu Lys Glu Phe Ala Ala Cys Pro Glu His Lys
210 215 220
Thr Ser Asp Ser Thr Phe Leu Val Phe Met Ser His Gly Leu Gln Glu
225 230 235 240
Gly Ile Cys Gly Ile Thr Tyr Ser Asn Glu Val Ala Asp Ile Leu Lys
245 250 255
Val Asp Thr Ile Phe Gln Met Met Asn Thr Leu Lys Cys Pro Ser Leu
260 265 270
Lys Asp Lys Pro Lys Val Ile Ile Ile Gln Ala Cys Arg Gly Glu Lys
275 280 285
Gln Gly Val Val Leu Leu Lys Asp Ser Val Gly Asn Ser Glu Glu Gly
290 295 300
Phe Leu Thr Asp Ala Ile Phe Glu Asp Asp Gly Ile Lys Lys Ala His
305 310 315 320
Ile Glu Lys Asp Phe Ile Ala Phe Cys Ser Ser Thr Pro Asp Asn Val
325 330 335
Ser Trp Arg His Pro Val Arg Gly Ser Leu Phe Ile Glu Ser Leu Ile
340 345 350
Lys His Met Lys Glu Tyr Ala Trp Ser Cys Asp Leu Glu Asp Ile Phe
355 360 365
Arg Lys Val Arg Phe Ser Phe Glu Gln Pro Asp Ser Arg Leu Gln Met
370 375 380
Pro Thr Thr Glu Arg Val Thr Leu Thr Lys Arg Phe Tyr Leu Phe Pro
385 390 395 400
Gly His
<210>334
<211>1209
<212>DNA
<213>Rattus norvegicus
<220>
<223>Caspase-1 precursor C14.001
<400>334
atggccgaca aggtcctgag ggcaaagagg aagcaattta tcaactcagt gagtgtaggg 60
acaataaatg gattgctgga tgaactttta gagaagagag tcctgaacca ggaagagatg 120
gatacaatca aacttgcaaa tatcactgtt atggaaaagg cacgagacct gtgcgatcat 180
gtcactaaaa aaggaccccg ggcaagccag atgtttatca cttacatttg taatgaagac 240
tgctacctgg cagaaattct ggagcttcag tcaggtccat cagctgaaac tgtttttgtt 300
acagaagatt ctaagggagg acatcctttc tcctcagaaa caaaagaaaa actgaacaaa 360
gaaggtggcg catttcctgg accgagtggt tccctcaagt tttgcccttt agaaatagcc 420
cagaaattat ggaaagaaaa tcattcagag atttatccaa taatgaaaac acccactcgt 480
acacgtcttg ccctcattat ctgcaacaca gactttcaac atctttctcg gagagtcgga 540
gctgatgttg acctcagaga aatgaagttg ctgctgcagg atctgggcta tactgtgaaa 600
gtcaaagaaa atctcacagc tctggagatg acaaaagaat tgaaagaatt tgctgcctgc 660
ccagagcaca agacttctga cagtaccttc cttgtattca tgtctcatgg tctccaggag 720
ggaatatgtg ggatcacata ctctaatgaa gttgcagata ttttaaaggt tgacacaatc 780
tttcaaatga tgaacacatt gaagtgccca agcttgaaag acaagcccaa ggttatcatt 840
attcaggcat gccgtggaga gaaacaagga gtggtgttgt taaaagattc agtaggaaac 900
tccgaagagg gattcttaac agatgcaatt tttgaagatg atggcattaa gaaggcccat 960
atagagaaag attttattgc tttctgctct tcaacaccag ataatgtgtc ctggagacat 1020
cctgtccggg gatccctctt cattgagtca ctcatcaaac acatgaaaga atatgcctgg 1080
tcttgtgact tggaggacat tttcagaaag gttcgatttt ctttcgaaca accagactct 1140
agactacaga tgccaaccac tgaaagggtg accctgacaa aacgtttcta cctcttccca 1200
ggacattag 1209
<210>335
<211>503
<212>PRT
<213>Caenorhabditis elegans
<220>
<223>Cell death protein 3 precursor C14.002
<400>335
Met Met Arg Gln Asp Arg Arg Ser Leu Leu Glu Arg Asn Ile Met Met
1 5 10 15
Phe Ser Ser His Leu Lys Val Asp Glu Ile Leu Glu Val Leu Ile Ala
20 25 30
Lys Gln Val Leu Asn Ser Asp Asn Gly Asp Met Ile Asn Ser Cys Gly
35 40 45
Thr Val Arg Glu Lys Arg Arg Glu Ile Val Lys Ala Val Gln Arg Arg
50 55 60
Gly Asp Val Ala Phe Asp Ala Phe Tyr Asp Ala Leu Arg Ser Thr Gly
65 70 75 80
His Glu Gly Leu Ala Glu Val Leu Glu Pro Leu Ala Arg Ser Val Asp
85 90 95
Ser Asn Ala Val Glu Phe Glu Cys Pro Met Ser Pro Ala Ser His Arg
100 105 110
Arg Ser Arg Ala Leu Ser Pro Ala Gly Tyr Thr Ser Pro Thr Arg Val
115 120 125
His Arp Asp Ser Val Ser Ser Val Ser Ser Phe Thr Ser Tyr Gln Asp
130 135 140
Ile Tyr Ser Arg Ala Arg Ser Arg Ser Arg Ser Arg Ala Leu His Ser
145 150 155 160
Ser Asp Arg His Asn Tyr Ser Ser Pro Pro Val Asn Ala Phe Pro Ser
165 170 175
Gln Pro Ser Ser Ala Asn Ser Ser Phe Thr Gly Cys Ser Ser Leu Gly
180 185 190
Tyr Ser Ser Ser Arg Asn Arg Ser Phe Ser Lys Ala Ser Gly Pro Thr
195 200 205
Gln Tyr Ile Phe His Glu Glu Asp Met Asn Phe Val Asp Ala Pro Thr
210 215 220
Ile Ser Arg Val Phe Asp Glu Lys Thr Met Tyr Arg Asn Phe Ser Ser
225 230 235 240
Pro Arg Gly Met Cys Leu Ile Ile Asn Asn Glu His Phe Glu Gln Met
245 250 255
Pro Thr Arg Asn Gly Thr Lys Ala Asp Lys Asp Asn Leu Thr Asn Leu
260 265 270
Phe Arg Cys Met Gly Tyr Thr Val Ile Cys Lys Asp Asn Leu Thr Gly
275 280 285
Arg Gly Met Leu Leu Thr Ile Arg Asp Phe Ala Lys His Glu Ser His
290 295 300
Gly Asp Ser Ala Ile Leu Val Ile Leu Ser His Gly Glu Glu Asn Val
305 310 315 320
Ile Ile Gly Val Asp Asp Ile Pro Ile Ser Thr His Glu Ile Tyr Asp
325 330 335
Leu Leu Asn Ala Ala Asn Ala Pro Arg Leu Ala Asn Lys Pro Lys Ile
340 345 350
Val Phe Val Gln Ala Cys Arg Gly Glu Arg Arg Asp Asn Gly Phe Pro
355 360 365
Val Leu Asp Ser Val Asp Gly Val Pro Ala Phe Leu Arg Arg Gly Trp
370 375 380
Asp Asn Arg Asp Gly Pro Leu Phe Asn Phe Leu Gly Cys Val Arg Pro
385 390 395 400
Gln Val Gln Gln Val Trp Arg Lys Lys Pro Ser Gln Ala Asp Ile Leu
405 410 415
Ile Ala Tyr Ala Thr Thr Ala Gln Tyr Val Ser Trp Arg Asn Ser Ala
420 425 430
Arg Gly Ser Trp Phe Ile Gln Ala Val Cys Glu Val Phe Ser Thr His
435 440 445
Ala Lys Asp Met Asp Val Val Glu Leu Leu Thr Glu Val Asn Lys Lys
450 455 460
Val Ala Cys Gly Phe Gln Thr Ser Gln Gly Ser Asn Ile Leu Lys Gln
465 470 475 480
Met Pro Glu Met Thr Ser Arg Leu Leu Lys Lys Phe Tyr Phe Trp Pro
485 490 495
Glu Ala Arg Asn Ser Ala Val
500
<210>336
<211>7653
<212>DNA
<213>Caenorhabditis elegans
<220>
<223>Cell death protein 3 precursor C14.002
<400>336
agatctgaaa taaggtgata aattaataaa ttaagtgtat ttctgaggaa atttgactgt 60
tttagcacaa ttaatcttgt ttcagaaaaa aagtccagtt ttctagattt ttccgtctta 120
ttgtcgaatt aatatcccta ttatcacttt ttcatgctca tcctcgagcg gcacgtcctc 180
aaagaattgt gagagcaaac gcgctcccat tgacctccac actcagccgc caaaacaaac 240
gttcgaacat tcgtgtgttg tgctcctttt ccgttatctt gcagtcatct tttgtcgttt 300
ttttctttgt tctttttgtt gaacgtgttg ctaagcaatt attacatcaa ttgaagaaaa 360
ggctcgccga tttattgttg ccagaaagat tctgagattc tcgaagtcga ttttataata 420
tttaaccttg gtttttgcat tgtttcgttt aaaaaaacca ctgtttatgt gaaaaacgat 480
tagtttacta ataaaactac ttttaaacct ttacctttac ctcaccgctc cgtgttcatg 540
gctcatagat tttcgatact caaatccaaa aataaattta cgagggcaat taatgtgaaa 600
caaaaacaat cctaagattt ccacatgttt gacctctccg gcaccttctt ccttagcccc 660
accactccat cacctctttg gcggtgttct tcgaaaccca cttaggaaag cagtgtgtat 720
ctcatttggt atgctctttt cgattttata gctctttgtc gcaatttcaa tgctttaaac 780
aatccaaatc gcattatatt tgtgcatgga ggcaaatgac ggggttggaa tcttagatga 840
gatcaggagc tttcagggta aacgcccggt tcattttgta ccacatttca tcattttcct 900
gtcgtccttg gtatcctcaa cttgtcccgg ttttgttttc ggtacactct tccgtgatgc 960
cacctgtctc cgtctcaatt atcgtttaga aatgtgaact gtccagatgg gtgactcata 1020
ttgctgctgc tacaatccac tttcttttct catcggcagt cttacgagcc catcataaac 1080
ttttttttcc gcgaaatttg caataaaccg gccaaaaact ttctccaaat tgttacgcaa 1140
tatatacaat ccataagaat atcttctcaa tgtttatgat ttcttcgcag cactttctct 1200
tcgtgtgcta acatcttatt tttataatat ttccgctaaa attccgattt ttgagtatta 1260
atttatcgta aaattatcat aatagcaccg aaaactacta aaaatggtaa aagctccttt 1320
taaatcggct cgacattatc gtattaagga atcacaaaat tctgagaatg cgtactgcgc 1380
aacatatttg acggcaaaat atctcgtagc gaaaactaca gtaattcttt aaatgactac 1440
tgtagcgctt gtgtcgattt acgggctcaa tttttgaaaa taattttttt tttcgaattt 1500
tgataacccg taaatcgtca caacgctaca gtagtcattt aaaggattac tgtagttcta 1560
gctacgagat attttgcgcg ccaaatatga ctgtaatacg cattctctga attttgtgtt 1620
tccgtaataa tttcacaaga ttttggcatt ccactttaaa ggcgcacagg atttattcca 1680
atgggtctcg gcacgcaaaa agtttgatag acttttaaat tctccttgca tttttaattc 1740
aattactaaa attttcgtga atttttctgt taaaattttt aaaatcagtt ttctaatatt 1800
ttccaggctg acaaacagaa acaaaaacac aacaaacatt ttaaaaatca gttttcaaat 1860
taaaaataac gatttctcat tgaaaattgt gttttatgtt tgcgaaaata aaagagaact 1920
gattcaaaac aattttaaca aaaaaaaacc ccaaaattcg ccagaaatca agataaaaaa 1980
ttcaagaggg tcaaaatttt ccgattttac tgactttcac cttttttttc gtagttcagt 2040
gcagttgttg gagtttttga cgaaaactag gaaaaaaatc gataaaaatt actcaaatcg 2100
agctgaattt tgaggacaat gtttaaaaaa aaacactatt tttccaataa tttcactcat 2160
tttcagacta aatcgaaaat caaatcgtac tctgactacg ggtcagtaga gaggtcaacc 2220
atcagccgaa gatgatgcgt caagatagaa ggagcttgct agagaggaac attatgatgt 2280
tctctagtca tctaaaagtc gatgaaattc tcgaagttct catcgcaaaa caagtgttga 2340
atagtgataa tggagatatg attaatgtga gtttttaatc gaataataat tttaaaaaaa 2400
aattgataat ataaagaata tttttgcagt catgtggaac ggttcgcgag aagagacggg 2460
agatcgtgaa agcagtgcaa cgacggggag atgtggcgtt cgacgcgttt tatgatgctc 2520
ttcgctctac gggacacgaa ggacttgctg aagttcttga acctctcgcc agatcgtagg 2580
tttttaaagt tcggcgcaaa agcaagggtc tcacggaaaa aagaggcgga tcgtaatttt 2640
gcaacccacc ggcacggttt tttcctccga aaatcggaaa ttatgcactt tcccaaatat 2700
ttgaagtgaa atatatttta tttactgaaa gctcgagtga ttatttattt tttaacacta 2760
attttcgtgg cgcaaaaggc cattttgtag atttgccgaa aatacttgtc acacacacac 2820
acacacatct ccttcaaata tccctttttc cagtgttgac tcgaatgctg tcgaattcga 2880
gtgtccaatg tcaccggcaa gccatcgtcg gagccgcgca ttgagccccg ccggctacac 2940
ttcaccgacc cgagttcacc gtgacagcgt ctcttcagtg tcatcattca cttcttatca 3000
ggatatctac tcaagagcaa gatctcgttc tcgatcgcgt gcacttcatt catcggatcg 3060
acacaattat tcatctcctc cagtcaacgc atttcccagc caaccttgta tgttgatgcg 3120
aacactaaat tctgagaatg cgcattactc aacatatttg acgcgcaaat atctcgtagc 3180
gaaaaataca gtaacccttt aaatgactat tgtagtgtcg atttacgggc tcgattttcg 3240
aaacgaatat atgctcgaat tgtgacaacg aattttaatt tgtcattttt gtgttttctt 3300
ttgatatttt tgatcaatta ataaattatt tccgtaaaca gacaccagcg ctacagtact 3360
cttttaaaga gttacagtag ttttcgcttc aagatatttt gaaaagaatt ttaaacattt 3420
tgaaaaaaaa tcatctaaca tgtgccaaaa cgcttttttc aagtttcgca gattttttga 3480
tttttttcat tcaagatatg cttattaaca catataatta tcattaatgt gaatttcttg 3540
tagaaatttt gggcttttcg ttctagtatg ctctactttt gaaattgctc aacgaaaaaa 3600
tcatgtggtt tgttcatatg aatgacgaaa aatagcaatt ttttatatat tttcccctat 3660
tcatgttgtg cagaaaaata gtaaaaaagc gcatgcattt ttcgacattt tttacatcga 3720
acgacagctc acttcacatg ctgaagacga gagacgcgga gaaataccac acatctttct 3780
gcgtctctcg tcttcagcat gtgaaatggg atctcggtcg atgtaaaaaa atgtcgaata 3840
atgtaaaaaa tgcatgcgtt tttttacact tttctgcaca aatgaatagg gggaaaatgt 3900
attaaaatac attttttgta tttttcaaca tcacatgatt aaccccatta ttttttcgtt 3960
gagcaactta aaaagtagag aatattagag cgaaaaccaa aatttcttca agatattacc 4020
tttattgata attatagatg ttaataagca tatcttgaat gaaagtcagc aaaaatatgt 4080
gcgaaacacc tgaaaaaaat caaaaattct gcgaaaattg aaaaaatgca ttaaaataca 4140
tttttgcatt tttctacatc acatgaatgt agaaaattaa aagggaaatc aaaatttcta 4200
gaggatataa ttgaatgaaa cattgcgaaa ttaaaatgtg cgaaacgtca aaaaagagga 4260
aatttgggta tcaaaatcga tcctaaaacc aacacatttc agcatccgcc aactcttcat 4320
tcaccggatg ctcttctctc ggatacagtt caagtcgtaa tcgctcattc agcaaagctt 4380
ctggaccaac tcaatacata ttccatgaag aggatatgaa ctttgtcgat gcaccaacca 4440
taagccgtgt tttcgacgag aaaaccatgt acagaaactt ctcgagtcct cgtggaatgt 4500
gcctcatcat aaataatgaa cactttgagc agatgccaac acggaatggt accaaggccg 4560
acaaggacaa tcttaccaat ttgttcagat gcatgggcta tacggttatt tgcaaggaca 4620
atctgacggg aagggtacgg cgaaattata ttacccaaac gcgaaatttg ccattttgcg 4680
ccgaaaatgt ggcgcccggt ctcgacacga caatttgtgt taaatgcaaa aatgtataat 4740
tttgcaaaaa acaaaatttt gaacttccgc gaaaatgatt tacctagttt cgaaattttc 4800
gttttttccg gctacattat gtgttttttc ttagtttttc tataatattt gatgtaaaaa 4860
accgtttgta aattttcaga caattttccg catacaaaac ttgatagcac gaaatcaatt 4920
ttctgaattt tcaaaattat ccaaaaatgc acaatttaaa atttgtgaaa attggcaaac 4980
ggtgtttcaa tatgaaatgt atttttaaaa actttaaaaa ccactccgga aaagcaataa 5040
aaatcaaaac aacgtcacaa ttcaaattca aaagttattc atccgatttg tttatttttg 5100
caaaatttga aaaaatcatg aaggatttag aaaagtttta taacattttt tctagatttt 5160
tcaaaatttt ttttaacaaa tcgagaaaaa gagaatgaaa aatcgatttt aaaaatatcc 5220
acagcttcga gagtttgaaa ttacagtact ccttaaaggc gcacacccca tttgcattgg 5280
accaaaaatt tgtcgtgtcg agaccaggta ccgtagtttt tgtcgcaaaa attgcaccat 5340
tggacaataa accttcctaa tcaccaaaaa gtaaaattga aatcttcgaa aagccaaaaa 5400
attcaaaaaa aaagtcgaat ttcgattttt tttttggttt tttggtccca aaaaccaaaa 5460
aaatcaattt tctgcaaaat accaaaaaga aacccgaaaa aatttcccag ccttgttcct 5520
aatgtaaact gatatttaat ttccagggaa tgctcctgac aattcgagac tttgccaaac 5580
acgaatcaca cggagattct gcgatactcg tgattctatc acacggagaa gagaatgtga 5640
ttattggagt tgatgatata ccgattagta cacacgagat atatgatctt ctcaacgcgg 5700
caaatgctcc ccgtctggcg aataagccga aaatcgtttt tgtgcaggct tgtcgaggcg 5760
gttcgttttt tattttaatt ttaatataaa tattttaaat aaattcattt tcagaacgtc 5820
gtgacaatgg attcccagtc ttggattctg tcgacggagt tcctgcattt cttcgtcgtg 5880
gatgggacaa tcgagacggg ccattgttca attttcttgg atgtgtgcgg ccgcaagttc 5940
aggttgcaat ttaatttctt gaatgagaat attccttcaa aaaatctaaa atagattttt 6000
attccagaaa gtcccgatcg aaaaattgcg atataattac gaaatttgtg ataaaatgac 6060
aaaccaatca gcatcgtcga tctccgccca cttcatcgga ttggtttgaa agtgggcgga 6120
gtgaattgct gattggtcgc agttttcagt ttagagggaa tttaaaaatc gccttttcga 6180
aaattaaaaa ttgatttttt caattttttc gaaaaatatt ccgattattt tatattcttt 6240
ggagcgaaag ccccgtcctg taaacatttt taaatgataa ttaataaatt tttgcagcaa 6300
gtgtggagaa agaagccgag ccaagctgac attctgattg catacgcaac gacagctcaa 6360
tatgtttcgt ggagaaacag tgctcgtgga tcatggttca ttcaagccgt ctgtgaagtg 6420
ttctcgacac acgcaaagga tatggatgtt gttgagctgc tgactgaagt caataagaag 6480
gtcgcttgtg gatttcagac atcacaggga tcgaatattt tgaaacagat gccagaggta 6540
cttgaaacaa acaatgcatg tctaactttt aaggacacag aaaaataggc agaggctcct 6600
tttgcaagcc tgccgcgcgt caacctagaa ttttagtttt tagctaaaat gattgatttt 6660
gaatatttta tgctaatttt tttgcgttaa attttgaaat agtcactatt tatcgggttt 6720
ccagtaaaaa atgtttatta gccattggat tttactgaaa acgaaaattt gtagtttttc 6780
aacgaaattt atcgattttt aaatgtaaaa aaaaatagcg aaaattacat caaccatcaa 6840
gcatttaagc caaaattgtt aactcattta aaaattaatt caaagttgtc cacgagtatt 6900
acacggttgg cgcgcggcaa gtttgcaaaa cgacgctccg cctctttttc tgtgcggctt 6960
gaaaacaagg gatcggttta gatttttccc caaaatttaa attaaatttc agatgacatc 7020
ccgcctgctc aaaaagttct acttttggcc ggaagcacga aactctgccg tctaaaattc 7080
actcgtgatt cattgcccaa ttgataattg tctgtatctt ctcccccagt tctctttcgc 7140
ccaattagtt taaaaccatg tgtatattgt tatcctatac tcatttcact ttatcattct 7200
atcatttctc ttcccatttt cacacatttc catttctcta cgataatcta aaattatgac 7260
gtttgtgtct cgaacgcata ataattttaa taactcgttt tgaatttgat tagttgttgt 7320
gcccagtata tatgtatgta ctatgcttct atcaacaaaa tagtttcata gatcatcacc 7380
ccaaccccac caacctaccg taccatattc atttttgccg ggaatcaatt tcgattaatt 7440
ttaacctatt ttttcgccac aaaaaatcta atatttgaat taacgaatag cattcccatc 7500
tctcccgtgc cggaatgcct cccggccttt taaagttcgg aacatttggc aattatgtat 7560
aaatttgtag gtccccccca tcatttcccg cccatcatct caaattgcat tcttttttcg 7620
ccgtgatatc ccgattctgg tcagcaaaga tct 7653
<210>337
<211>277
<212>PRT
<213>Homo sapiens
<220>
<223>Caspase-3 precursor C14.003
<400>337
Met Glu Asn Thr Glu Asn Ser Val Asp Ser Lys Ser Ile Lys Asn Leu
1 5 10 15
Glu Pro Lys Ile Ile His Gly Ser Glu Ser Met Asp Ser Gly Ile Ser
20 25 30
Leu Asp Asn Ser Tyr Lys Met Asp Tyr Pro Glu Met Gly Leu Cys Ile
35 40 45
Ile Ile Asn Asn Lys Asn Phe His Lys Ser Thr Gly Met Thr Ser Arg
50 55 60
Ser Gly Thr Asp Val Asp Ala Ala Asn Leu Arg Glu Thr Phe Arg Asn
65 70 75 80
Leu Lys Tyr Glu Val Arg Asn Lys Asn Asp Leu Thr Arg Glu Glu Ile
85 90 95
Val Glu Leu Met Arg Asp Val Ser Lys Glu Asp His Ser Lys Arg Ser
100 105 110
Ser Phe Val Cys Val Leu Leu Ser His Gly Glu Glu Gly Ile Ile Phe
115 120 125
Gly Thr Asn Gly Pro Val Asp Leu Lys Lys Ile Thr Asn Phe Phe Arg
130 135 140
Gly Asp Arg Cys Arg Ser Leu Thr Gly Lys Pro Lys Leu Phe Ile Ile
145 150 155 160
Gln Ala Cys Arg Gly Thr Glu Leu Asp Cys Gly Ile Glu Thr Asp Ser
165 170 175
Gly Val Asp Asp Asp Met Ala Cys His Lys Ile Pro Val Glu Ala Asp
180 185 190
Phe Leu Tyr Ala Tyr Ser Thr Ala Pro Gly Tyr Tyr Ser Trp Arg Asn
195 200 205
Ser Lys Asp Gly Ser Trp Phe Ile Gln Ser Leu Cys Ala Met Leu Lys
210 215 220
Gln Tyr Ala Asp Lys Leu Glu Phe Met His Ile Leu Thr Arg Val Asn
225 230 235 240
Arg Lys Val Ala Thr Glu Phe Glu Ser Phe Ser Phe Asp Ala Thr Phe
245 250 255
His Ala Lys Lys Gln Ile Pro Cys Ile Val Ser Met Leu Thr Lys Glu
260 265 270
Leu Tyr Phe Tyr His
275
<210>338
<211>2635
<212>DNA
<213>Homo sapiens
<220>
<223>Caspase-3 precursor C14.003
<400>338
gaattcggca cgaggggtgc tattgtgagg cggttgtaga agagtttcgt gagtgctcgc 60
agctcatacc tgtggctgtg tatccgtggc cacagctggt tggcgtcgcc ttgaaatccc 120
aggccgtgag gagttagcga gccctgctca cactcggcgc tctggttttc ggtgggtgtg 180
ccctgcacct gcctcttccc ccattctcat taataaaggt atccatggag aacactgaaa 240
actcagtgga ttcaaaatcc attaaaaatt tggaaccaaa gatcatacat ggaagcgaat 300
caatggactc tggaatatcc ctggacaaca gttataaaat ggattatcct gagatgggtt 360
tatgtataat aattaataat aagaattttc ataaaagcac tggaatgaca tctcggtctg 420
gtacagatgt cgatgcagca aacctcaggg aaacattcag aaacttgaaa tatgaagtca 480
ggaataaaaa tgatcttaca cgtgaagaaa ttgtggaatt gatgcgtgat gtttctaaag 540
aagatcacag caaaaggagc agttttgttt gtgtgcttct gagccatggt gaagaaggaa 600
taatttttgg aacaaatgga cctgttgacc tgaaaaaaat aacaaacttt ttcagagggg 660
atcgttgtag aagtctaact ggaaaaccca aacttttcat tattcaggcc tgccgtggta 720
cagaactgga ctgtggcatt gagacagaca gtggtgttga tgatgacatg gcgtgtcata 780
aaataccagt ggatgccgac ttcttgtatg catactccac agcacctggt tattattctt 840
ggcgaaattc aaaggatggc tcctggttca tccagtcgct ttgtgccatg ctgaaacagt 900
atgccgacaa gcttgaattt atgcacattc ttacccgggt taaccgaaag gtggcaacag 960
aatttgagtc cttttccttt gacgctactt ttcatgcaaa gaaacagatt ccatgtattg 1020
tttccatgct cacaaaagaa ctctattttt atcactaaag aaatggttgg ttggtggttt 1080
tttttagttt gtatgccaag tgagaagatg gtatatttgg tactgtattt ccctctcatt 1140
ttgacctact ctcatgctgc agagggtact ttaagacata ctccttccat caaatagaac 1200
cactatgaag ctacctcaaa cttccagtca ggtagttgca attgaattaa attaggaata 1260
aataaaaatg gatactggtg cagtcattat gagaggcaat gattgttaat ttacagcttt 1320
catgattagc aagttacagt gatgctgtgc tatgaatttt caagtaattg tgaaaaagtt 1380
aaacattgaa gtaatgaatt tttatgatat tccccccact taagactgtg tattctagtt 1440
ttgtcaaact gtagaaatga tgatgtggaa gaacttaggc atctgtgggc atggtcaaag 1500
gctcaaacct ttattttaga attgatatac acggatgact taactgcatt ttagaccatt 1560
tatctgggat tatggttttg tgatgtttgt cctgaacact tttgttgtaa aaaaataata 1620
ataataatgt ttaatattga gaaagaaact aatattttat gtgagagaaa gtgtgagcaa 1680
actaacttga cttttaaggc taaaacttaa cattcataga ggggtggagt tttaactgta 1740
aggtgctaca atgcccctgg atctaccagc ataaatatct tctgatttgt ccctatgcat 1800
atcagttgag cttcatatac cagcaatata tctgaagagc tattatataa aaaccccaaa 1860
ctgttgatta ttagccaggt aatgtgaata aattctatag gaacatatga aaatacaact 1920
taaataataa acagtggaat ataaggaaag caataaatga atgggctgag ctgcctgtaa 1980
cttgagagta gatggtttga gcctgagcag agacatgact cagcctgttc catgaaggca 2040
gagccatgga ccacgcagga agggcctaca gcccatttct ccatacgcac tggtatgtgt 2100
ggatgatgct gccagggcgc catcgccaag taagaaagtg aagcaaatca gaaacttgtg 2160
aagtggaaat gttctaaagg tggtgaggca ataaaaatca tagtactctt tgtagcaaaa 2220
ttcttaagta tgttattttc tgttgaagtt tacaatcaaa ggaaaatagt aatgttttat 2280
actgtttact gaaagaaaaa gacctatgag cacataggac tctagacggc atccagccgg 2340
aggccagagc tgagcactca gcccgggagg caggctccag gcctcagcag gtgcggagcc 2400
gtcactgcac caagtctcac tggctgtcag tatgacattt cacgggagat ttcttgttgc 2460
tcaaaaaatg agctcgcatt tgtcaatgac agtttctttt ttcttactag acctgtaact 2520
tttgtaaata cacacagcat gtaatggtat cttaaagtgt gtttctatgt gacaattttg 2580
tacaaatttg ttattttcca tttttatttc aaaatataca ttcaaactta aaatt 2635
<210>339
<211>303
<212>PRT
<213>Homo sapiens
<220>
<400>339
Met Ala Asp Asp Gln Gly Cys Ile Glu Glu Gln Gly Val Glu Asp Ser
1 5 10 15
Ala Asn Glu Asp Ser Val Asp Ala Lys Pro Asp Arg Ser Ser Phe Val
20 25 30
Pro Ser Leu Phe Ser Lys Lys Lys Lys Asn Val Thr Met Arg Ser Ile
35 40 45
Lys Thr Thr Arg Asp Arg Val Pro Thr Tyr Gln Tyr Asn Met Asn Phe
50 55 60
Glu Lys Leu Gly Lys Cys Ile Ile Ile Asn Asn Lys Asn Phe Asp Lys
65 70 75 80
Val Thr Gly Met Gly Val Arg Asn Gly Thr Asp Lys Asp Ala Glu Ala
85 90 95
Leu Phe Lys Cys Phe Arg Ser Leu Gly Phe Asp Val Ile Val Tyr Asn
100 105 110
Asp Cys Ser Cys Ala Lys Met Gln Asp Leu Leu Lys Lys Ala Ser Glu
115 120 125
Glu Asp His Thr Asn Ala Ala Cys Phe Ala Cys Ile Leu Leu Ser His
130 135 140
Gly Glu Glu Asn Val Ile Tyr Gly Lys Asp Gly Val Thr Pro Ile Lys
145 150 155 160
Asp Leu Thr Ala His Phe Arg Gly Asp Arg Cys Lys Thr Leu Leu Glu
165 170 195
Lys Pro Lys Leu Phe Phe Ile Gln Ala Cys Arg Gly Thr Glu Leu Asp
180 185 190
Asp Gly Ile Gln Ala Asp Ser Gly Pro Ile Asn Asp Thr Asp Ala Asn
195 200 205
Pro Arg Tyr Lys Ile Pro Val Glu Ala Asp Phe Leu Phe Ala Tyr Ser
210 215 220
Thr Val Pro Gly Tyr Tyr Ser Trp Arg Ser Pro Gly Arg Gly Ser Trp
225 230 235 240
Phe Val Gln Ala Leu Cys Ser Ile Leu Glu Glu His Gly Lys Asp Leu
245 250 255
Glu Ile Met Gln Ile Leu Thr Arg Val Asn Asp Arg Val Ala Arg His
260 265 270
Phe Glu Ser Gln Ser Asp Asp Pro His Phe His Glu Lys Lys Gln Ile
275 280 285
Pro Cys Val Val Ser Met Leu Thr Lys Glu Leu Tyr Phe Ser Gln
290 295 300
<210>340
<211>912
<212>DNA
<213>Homo sapiens
<220>
<223>Caspase-7 precursor C14.004
<400>340
atggcagatg atcagggctg tattgaagag cagggggttg aggattcagc aaatgaagat 60
tcagtggatg ctaagccaga ccggtcctcg tttgtaccgt ccctcttcag taagaagaag 120
aaaaatgtca ccatgcgatc catcaagacc acccgggacc gagtgcctac atatcagtac 180
aacatgaatt ttgaaaagct gggcaaatgc atcataataa acaacaagaa ctttgataaa 240
gtgacaggta tgggcgttcg aaacggaaca gacaaagatg ccgaggcgct cttcaagtgc 300
ttccgaagcc tgggttttga cgtgattgtc tataatgact gctcttgtgc caagatgcaa 360
gatctgctta aaaaagcttc tgaagaggac catacaaatg ccgcctgctt cgcctgcatc 420
ctcttaagcc atggagaaga aaatgtaatt tatgggaaag atggtgtcac accaataaag 480
gatttgacag cccactttag gggggataga tgcaaaaccc ttttagagaa acccaaactc 540
ttcttcattc aggcttgccg agggaccgag cttgatgatg ccatccaggc cgactcgggg 600
cccatcaatg acacagatgc taatcctcga tacaagatcc cagtggaagc tgacttcctc 660
ttcgcctatt ccacggttcc aggctattac tcgtggagga gcccaggaag aggctcctgg 720
tttgtgcaag ccctctgctc catcctggag gagcacggaa aagacctgga aatcatgcag 780
atcctcacca gggtgaatga cagagttgcc aggcactttg agtctcagtc tgatgaccca 840
cacttccatg agaagaagca gatcccctgt gtggtctcca tgctcaccaa ggaactctac 900
ttcagtcaat ag 912
<210>341
<211>293
<212>PRT
<213>Homo sapiens
<220>
<223>Caspase-6 precursor C14.005
<400>341
Met Ser Ser Ala Ser Gly Leu Arg Arg Gly His Pro Ala Gly Gly Glu
1 5 10 15
Glu Asn Met Thr Glu Thr Asp Ala Phe Tyr Lys Arg Glu Met Phe Asp
20 25 30
Pro Ala Glu Lys Tyr Lys Met Asp His Arg Arg Arg Gly Ile Ala Leu
35 40 45
Ile Phe Asn His Glu Arg Phe Phe Trp His Leu Thr Leu Pro Glu Arg
50 55 60
Arg Gly Thr Cys Ala Asp Arg Asp Asn Leu Thr Arg Arg Phe Ser Asp
65 70 75 80
Leu Gly Phe Glu Val Lys Cys Phe Asn Asp Leu Lys Ala Glu Glu Leu
85 90 95
Leu Leu Lys Ile His Glu Val Ser Thr Val Ser His Ala Asp Ala Asp
100 105 110
Cys Phe Val Cys Val Phe Leu Ser His Gly Glu Gly Asn His Ile Tyr
115 120 125
Ala Tyr Asp Ala Lys Ile Glu Ile Gln Thr Leu Thr Gly Leu Phe Lys
130 135 140
Gly Asp Lys Cys His Ser Leu Val Gly Lys Pro Lys Ile Phe Ile Ile
145 150 155 160
Gln Ala Cys Arg Gly Asn Gln His Asp Val Pro Val Ile Pro Leu Asp
165 170 175
Val Val Asp Asn Gln Thr Glu Lys Leu Asp Thr Asn Ile Thr Glu Val
180 185 190
Asp Ala Ala Ser Val Tyr Thr Leu Pro Ala Gly Ala Asp Phe Leu Met
195 200 205
Cys Tyr Ser Val Ala Glu Gly Tyr Tyr Ser His Arg Glu Thr Val Asn
210 215 220
Gly Ser Trp Tyr Ile Gln Asp Leu Cys Glu Met Leu Gly Lys Tyr Gly
225 230 235 240
Ser Ser Leu Glu Phe Thr Glu Leu Leu Thr Leu Val Asn Arg Lys Val
245 250 255
Ser Gln Arg Arg Val Asp Phe Cys Lys Asp Pro Ser Ala Ile Gly Lys
260 265 270
Lys Gln Val Pro Cys Phe Ala Ser Met Leu Thr Lys Lys Leu His Phe
275 280 285
Phe Pro Lys Ser Asn
290
<210>342
<211>1545
<212>DNA
<213>Homo sapiens
<220>
<223>Caspase-6 precursor C14.005
<400>342
ccgagggcgg ggccgggccc gggagcctgt ggcttcagga agaggagggc aaggtgtctg 60
gctgcgcgtt tggctgcaat gagctcggcc tcggggctcc gcagggggca cccggcaggt 120
ggggaagaaa acatgacaga aacagatgcc ttctataaaa gagaaatgtt tgatccggca 180
gaaaagtaca aaatggacca caggaggaga ggaattgctt taatcttcaa tcatgagagg 240
ttcttttggc acttaacact gccagaaagg cggcgcacct gcgcagatag agacaatctt 300
acccgcaggt tttcagatct aggatttgaa gtgaaatgct ttaatgatct taaagcagaa 360
gaactactgc tcaaaattca tgaggtgtca actgttagcc acgcagatgc cgattgcttt 420
gtgtgtgtct tcctgagcca tggcgaaggc aatcacattt atgcatatga tgctaaaatc 480
gaaattcaga cattaactgg cttgttcaaa ggagacaagt gtcacagcct ggttggaaaa 540
cccaagatat ttatcatcca ggcatgtcgg ggaaaccagc acgatgtgcc agtcattcct 600
ttggatgtag tagataatca gacagagaag ttggacacca acataactga ggtggatgca 660
gcctccgttt acacgctgcc tgctggagct gacttcctca tgtgttactc tgttgcagaa 720
ggatattatt ctcaccggga aactgtgaac ggctcatggt acattcaaga tttgtgtgag 780
atgttgggaa aatatggctc ctccttagag ttcacagaac tcctcacact ggtgaacagg 840
aaagtttctc agcgccgagt ggacttttgc aaagacccaa gtgcaattgg aaagaagcag 900
gttccctgtt ttgcctcaat gctaactaaa aagctgcatt tctttccaaa atctaattaa 960
ttaatagagg ctatctaatt tcacactctg tattgaaaat ggctttctca gccaggcgtg 1020
gttactcaca cctgtaatcc cagcactttg ggagtccaag gtgggcggat cacctgaggt 1080
cgggagttcg agaccagcct gaccaacatg gcagaagccc cgcctctact aaaaatgcaa 1140
aaaaaaattt agctaggcat ggcggcgcat gcctgcaatc ccagctactt ggaaggctga 1200
ggcaggagaa tcacttgaac ccaggaggtg gaggctgcgg tgagccgagc attgcgccat 1260
tgcactccag cctgggcaac gagtgaaact ccgtctcaaa aaaaaagaaa atgtctttct 1320
cttcctttta tataaatatc gttagggtga agcattatgg tctaatgatt caaatgtttt 1380
aaagtttaat gcctagcaga gaactgcctt aaaaaaaaaa agttcatgtt ggccatggtg 1440
aaagggtttg atatggagaa acaaaatcct caggaaatta gataaataaa aatttataag 1500
catttgtatt attttttaat aaactgcagg gttacacaaa aatct 1545
<210>343
<211>452
<212>PRT
<213>Mus musculus
<220>
<223>Caspase-2 precursor C14.006
<400>343
Met Ala Ala Pro Ser Gly Arg Ser Gln Ser Ser Leu His Arg Lys Gly
1 5 10 15
Leu Met Ala Ala Asp Arg Arg Ser Arg Ile Leu Ala Val Cys Gly Met
20 25 30
His Pro Asp His Gln Glu Thr Leu Lys Lys Asn Arg Val Val Leu Ala
35 40 45
Lys Gln Leu Leu Leu Ser Glu Leu Leu Glu His Leu Leu Glu Lys Asp
50 55 60
Ile Ile Thr Leu Glu Met Arg Glu Leu Ile Gln Ala Lys Gly Gly Ser
65 70 75 80
Phe Ser Gln Asn Val Glu Leu Leu Asn Leu Leu Pro Lys Arg Gly Pro
85 90 95
Gln Ala Phe Asp Ala Phe Cys Glu Ala Leu Arg Glu Thr Arg Gln Gly
100 105 110
His Leu Glu Asp Leu Leu Leu Thr Thr Leu Ser Asp Ile Gln His Val
115 120 125
Leu Pro Pro Leu Ser Cys Asp Tyr Asp Thr Ser Leu Pro Phe Ser Val
130 135 140
Cys Glu Ser Cys Pro Pro His Lys Gln Leu Arg Leu Ser Thr Asp Ala
145 150 155 160
Thr Glu His Ser Leu Asp Asn Gly Asp Gly Pro Pro Cys Leu Leu Val
165 170 175
Lys Pro Cys Thr Pro Glu Phe Tyr Gln Ala His Tyr Gln Leu Ala Tyr
180 185 190
Arg Leu Gln Ser Gln Pro Arg Gly Leu Ala Leu Val Leu Ser Asn Val
195 200 205
His Phe Thr Gly Glu Lys Asp Leu Glu Phe Arg Ser Gly Gly Asp Val
210 215 220
Asp His Thr Thr Leu Val Thr Leu Phe Lys Leu Leu Gly Tyr Asn Val
225 230 235 240
His Val Leu His Asp Gln Thr Ala Gln Glu Met Gln Glu Lys Leu Gln
245 250 255
Asn Phe Ala Gln Leu Pro Ala His Arg Val Thr Asp Ser Cys Val Val
260 265 270
Ala Leu Leu Ser His Gly Val Glu Gly Gly Ile Tyr Gly Val Asp Gly
275 280 285
Lys Leu Leu Gln Leu Gln Glu Val Phe Arg Leu Phe Asp Asn Ala Asn
290 295 300
Cys Pro Ser Leu Gln Asn Lys Pro Lys Met Phe Phe Ile Gln Ala Cys
305 310 315 320
Arg Gly Asp Glu Thr Asp Arg Gly Val Asp Gln Gln Asp Gly Lys Asn
325 330 335
His Thr Gln Ser Pro Gly Cys Glu Glu Ser Asp Ala Gly Lys Glu Glu
340 345 350
Leu Met Lys Met Arg Leu Pro Thr Arg Ser Asp Met Ile Cys Gly Tyr
355 360 365
Ala Cys Leu Lys Gly Asn Ala Ala Met Arg Asn Thr Lys Arg Gly Ser
370 375 380
Trp Tyr Ile Glu Ala Leu Thr Gln Val Phe Ser Glu Arg Ala Cys Asp
385 390 395 400
Met His Val Ala Asp Met Leu Val Lys Val Asn Ala Leu Ile Lys Glu
405 410 415
Arg Glu Gly Tyr Ala Pro Gly Thr Glu Phe His Arg Cys Lys Glu Met
420 425 430
Ser Glu Tyr Cys Ser Thr Leu Cys Gln Gln Leu Tyr Leu Phe Pro Gly
435 440 445
Tyr Pro Pro Thr
450
<210>344
<211>3395
<212>DNA
<213>Mus musculus
<220>
<223>Caspase-2 precursor C14.006
<400>344
ctggaaatgg cggcgccgag cgggaggtcg cagtcctccc tgcacaggaa ggggctgatg 60
gcggctgaca ggaggagcag gattttggca gtgtgtggaa tgcatcctga ccaccaggaa 120
acactgaaaa agaatcgagt ggtgctggcc aagcagctgc tgctgagcga gctgttagaa 180
cacctcctag agaaggacat tatcactttg gaaatgaggg agctcatcca ggccaaaggg 240
ggcagtttca gccagaatgt ggaactcctc aacctgctgc caaagagagg accccaggct 300
tttgatgcct tctgtgaagc cctgcgggag accaggcagg gtcacttgga agacttactg 360
ctcacaaccc tctcagatat tcagcacgta ctcccaccgt tgagctgtga ctatgacaca 420
agtctccctt tctcggtgtg tgagtcctgc cctcctcaca agcagctccg cctatccaca 480
gatgctacgg aacactcctt agataatggt gatggtcctc cctgtcttct ggtgaagcca 540
tgcactcctg agttttacca ggcacactac cagctggcct ataggttgca atctcagccc 600
cgtggcttgg cactggtgct gagcaatgtg cacttcactg gagagaaaga cctggaattc 660
cgctctggag gggatgtgga ccacactact ctagtcaccc tcttcaagct tttgggctac 720
aatgtccatg tgctacatga ccagaccgca caggaaatgc aagagaaact tcagaatttt 780
gcacagttac ctgcacaccg ggtcacagac tcctgcgtag tggcactcct ctcacatggt 840
gtggaaggtg gcatctatgg tgtagatggc aaactgcttc agctccaaga ggtttttcga 900
ctttttgaca atgctaactg tccaagtcta cagaacaagc caaaaatgtt cttcatccaa 960
gcatgtcgtg gagatgagac agatagaggt gtcgaccagc aagatggaaa gaaccacaca 1020
caatcccctg gatgtgagga gagtgatgct ggcaaagagg agttgatgaa gatgagactg 1080
cctactcgct cagacatgat atgtggctat gcttgcctta aaggtaatgc tgccatgcgg 1140
aacaccaaac ggggttcctg gtacattgag gccctcactc aggtgttctc tgaaagagct 1200
tgtgacatgc acgtggccga catgcttgtt aaggtgaatg cccttatcaa ggagcgtgaa 1260
ggctatgccc ctggcacaga attccaccga tgcaaggaga tgtctgagta ctgtagtact 1320
ctgtgccagc aactctacct gttcccaggc tacccaccca cgtgatgccg cctgctattc 1380
ctgctgttgg aggccactgg accactgggg gcacaatgga gacttctctt cagaatggtt 1440
tttgttctgt ctaccctctc agggatatga gattctccca ggcttgtttc ctgtcagcca 1500
tctctgtctt tgggtatgaa acataaggat ggctcctccg gtgtcgtgtt ctctacctat 1560
agagccagct ctgaatggat gtgttaccag aagcatttta gctacagcct agaaaatgac 1620
attgtgaaca cagtattatt gtgggaagag ggcatttgga tttctcaatg tttgtgatat 1680
ttttgttccc aaggcatctt aggagtactt ggatcatagc tttttttttt tttcctaaat 1740
cagttaagga gtctcagaga tcatctcctt ttttttccat atctacaacc tcatttttcc 1800
cacagtggag atttggaaga tgtcccaatt taatgtaggt gttttcatct gtcatgaagg 1860
gacagatgag atcctactac ttgcgaagtt tctatgcata cctttaagtt caggccctag 1920
gtgaaggaca gtccctcagc ctttccattg gttcctttgt gttcagtgca cccagccttt 1980
gaacagagcc tagggtctgt atgccatgac actggaagtc atagaaattt ccctggtcat 2040
gctttgtttg aactgtcact gaatgaacct tatcgggcat aactacatga aaatgcagtg 2100
acagctgagt gtgctgtgtc tcacactatc acccgtcatc aggatgtctc tccttcctta 2160
ctgtggcttc tgcatgcact tacactgtac ttgacggctg gcctccaggg tctctcttgc 2220
tttgtactgg ttcccctctt taccttcacc attcgctgct tctgccaagt ctgtgaagcc 2280
gtcctttgta ggatgtttct tgccacttac gctgtactgt agttgcttat tctttctgcc 2340
ttctgcttca gcgtgaggct tctttggttt tctgtggcag cgtctccctt ctcattgttt 2400
ctctgtgttt tagtggggat agtaccatat gtgatataac ctagaagcac ttgtctctgc 2460
tcttatgaaa cttgcttatt cttgaaaacc ttctgcattt ccattttttc ctctcttcca 2520
atttattctc catgtaacag agtagtttgg tttttaaaat atctggtgat gtcattctct 2580
tgcttagaac actagcttcc tgttacgctt catctaaaat gcaaattctt acacccagct 2640
tacgagatct ggctcatacc ttccctttgg atctcattaa atggtgattt ctcactatgc 2700
tccagcccct cttaggtcct catctccgtc ttgcaggtgt tctgaactct cctttggcta 2760
gtctctgatt tttgagtctg gcggaggcct cttgaccatg ctgcccatgc tgtctactgt 2820
gcctccttat gagggcatca tgttggtctc tgttgtgctt actgcaggct gtaatggcac 2880
ttttgcttgt ttcacttgtt ccctctgagg ctgaatgctc caagagagtg ggaactgtgc 2940
ttcttactta ctgatatcca gtaactggca cttactaggt cttcatgaat gtttcctgag 3000
taaaggaagg agaccagcag ctaaccttag ttagagccta ccttttgcag tttctaaatt 3060
gctattatag tgtacagttc aattagtata tgggtttttt tttccaggtg ttttattttt 3120
atccactgtt ttgttgttgt ttttttatat tttctacata tcacgtgtga gacaacttct 3180
ttcacatctc catagtgccc agcaaatttg aggcctatgg tagttgaggt gctcaactaa 3240
tgtttgttgt atgaaccaag tggtttgaag acttgctgcc aaattctgcc ttttgggtca 3300
gtataggatg cataagtggt agaatcttca cacttcccac tgccaagatt ttgtattgcc 3360
atcaggtgcc aaataaatgt tgatacttat tactg 3395
<210>345
<211>377
<212>PRT
<213>Homo sapiens
<220>
<223>Caspase-4 precursor C14.007
<400>345
Met Ala Glu Gly Asn His Arg Lys Lys Pro Leu Lys Val Leu Glu Ser
1 5 10 15
Leu Gly Lys Asp Phe Leu Thr Gly Val Leu Asp Asn Leu Val Glu Gln
20 25 30
Asn Val Leu Asn Trp Lys Glu Glu Glu Lys Lys Lys Tyr Tyr Asp Ala
35 40 45
Lys Thr Glu Asp Lys Val Arg Val Met Ala Asp Ser Met Gln Glu Lys
50 55 60
Gln Arg Met Ala Gly Gln Met Leu Leu Gln Thr Phe Phe Asn Ile Asp
65 70 75 80
Gln Ile Ser Pro Asn Lys Lys Ala His Pro Asn Met Glu Ala Gly Pro
85 90 95
Pro Glu Ser Gly Glu Ser Thr Asp Ala Leu Lys Leu Cys Pro His Glu
100 105 110
Glu Phe Leu Arg Leu Cys Lys Glu Arg Ala Glu Glu Ile Tyr Pro Ile
115 120 125
Lys Glu Arg Asn Asn Arg Thr Arg Leu Ala Leu Ile Ile Cys Asn Thr
130 135 140
Glu Phe Asp His Leu Pro Pro Arg Asn Gly Ala Asp Phe Asp Ile Thr
145 150 155 160
Gly Met Lys Glu Leu Leu Glu Gly Leu Asp Tyr Ser Val Asp Val Glu
165 170 175
Glu Asn Leu Thr Ala Arg Asp Met Glu Ser Ala Leu Arg Ala Phe Ala
180 185 190
Thr Arg Pro Glu His Lys Ser Ser Asp Ser Thr Phe Leu Val Leu Met
195 200 205
Ser His Gly Ile Leu Glu Gly Ile Cys Gly Thr Val His Asp Glu Lys
210 215 220
Lys Pro Asp Val Leu Leu Tyr Asp Thr Ile Phe Gln Ile Phe Asn Asn
225 230 235 240
Arg Asn Cys Leu Ser Leu Lys Asp Lys Pro Lys Val Ile Ile Val Gln
245 250 255
Ala Cys Arg Gly Ala Asn Arg Gly Glu Leu Trp Val Arg Asp Ser Pro
260 265 270
Ala Ser Leu Glu Val Ala Ser Ser Gln Ser Ser Glu Asn Leu Glu Glu
275 280 285
Asp Ala Val Tyr Lys Thr His Val Glu Lys Asp Phe Ile Ala Phe Cys
290 295 300
Ser Ser Thr Pro His Asn Val Ser Trp Arg Asp Ser Thr Met Gly Ser
305 310 315 320
Ile Phe Ile Thr Gln Leu Ile Thr Cys Phe Gln Lys Tyr Ser Trp Cys
325 330 335
Cys His Leu Glu Glu Val Phe Arg Lys Val Gln Gln Ser Phe Glu Thr
340 345 350
Pro Arg Ala Lys Ala Gln Met Pro Thr Ile Glu Arg Leu Ser Met Thr
355 360 365
Arg Tyr Phe Tyr Leu Phe Pro Gly Asn
370 375
<210>346
<211>1291
<212>DNA
<213>Homo sapiens
<220>
<223>Caspase-4 precursor C14.007
<400>346
gctctttcca acgctgtaaa aaaggacaga ggctgttccc tatggcagaa ggcaaccaca 60
gaaaaaagcc acttaaggtg ttggaatccc tgggcaaaga tttcctcact ggtgttttgg 120
ataacttggt ggaacaaaat gtactgaact ggaaggaaga ggaaaaaaag aaatattacg 180
atgctaaaac tgaagacaaa gttcgggtca tggcagactc tatgcaagag aagcaacgta 240
tggcaggaca aatgcttctt caaacctttt ttaacataga ccaaatatcc cccaataaaa 300
aagctcatcc gaatatggag gctggaccac ctgagtcagg agaatctaca gatgccctca 360
agctttgtcc tcatgaagaa ttcctgagac tatgtaaaga aagagctgaa gagatctatc 420
caataaagga gagaaacaac cgcacacgcc tggctctcat catatgcaat acagagtttg 480
accatctgcc tccgaggaat ggagctgact ttgacatcac agggatgaag gagctacttg 540
agggtctgga ctatagtgta gatgtagaag agaatctgac agccagggat atggagtcag 600
cgctgagggc atttgctacc agaccagagc acaagtcctc tgacagcaca ttcttggtac 660
tcatgtctca tggcatcctg gagggaatct gcggaactgt gcatgatgag aaaaaaccag 720
atgtgctgct ttatgacacc atcttccaga tattcaacaa ccgcaactgc ctcagtctga 780
aggacaaacc caaggtcatc attgtccagg cctgcagagg tgcaaaccgt ggggaactgt 840
gggtcagaga ctctccagca tccttggaag tggcctcttc acagtcatct gagaacctgg 900
aggaagatgc tgtttacaag acccacgtgg agaaggactt cattgctttc tgctcttcaa 960
cgccacacaa cgtgtcctgg agagacagca caatgggctc tatcttcatc acacaactca 1020
tcacatgctt ccagaaatat tcttggtgct gccacctaga ggaagtattt cggaaggtac 1080
agcaatcatt tgaaactcca agggccaaag ctcaaatgcc caccatagaa cgactgtcca 1140
tgacaagata tttctacctc tttcctggca attgaaaatg gaagccacaa gcagcccagc 1200
cctccttaat caacttcaag gagcaccttc attagtacag cttgcatatt taacattttg 1260
tatttcaata aaagtgaaga caaaaaaaaa a 1291
<210>347
<211>418
<212>PRT
<213>Homo sapiens
<220>
<223>Caspase-5 precursor C14.008
<400>347
Met Phe Lys Gly Ile Leu Gln Ser Gly Leu Asp Asn Phe Val Ile Asn
1 5 10 15
His Met Leu Lys Asn Asn Val Ala Gly Gln Thr Ser Ile Gln Thr Leu
20 25 30
Val Pro Asn Thr Asp Gln Lys Ser Thr Ser Val Lys Lys Asp Asn His
35 40 45
Lys Lys Lys Thr Val Lys Met Leu Glu Tyr Leu Gly Lys Asp Val Leu
50 55 60
His Gly Val Phe Asn Tyr Leu Ala Lys His Asp Val Leu Thr Leu Lys
65 70 75 80
Glu Glu Glu Lys Lys Lys Tyr Tyr Asp Ala Lys Ile Glu Asp Lys Ala
85 90 95
Leu Ile Leu Val Asp Ser Leu Arg Lys Asn Arg Val Ala His Gln Met
100 105 110
Phe Thr Gln Thr Leu Leu Asn Met Asp Gln Lys Ile Thr Ser Val Lys
115 120 125
Pro Leu Leu Gln Ile Glu Ala Gly Pro Pro Glu Ser Ala Glu Ser Thr
130 135 140
Asn Ile Leu Lys Leu Cys Pro Arg Glu Glu Phe Leu Arg Leu Cys Lys
145 150 155 160
Lys Asn His Asp Glu Ile Tyr Pro Ile Lys Lys Arg Glu Asp Arg Arg
165 170 175
Arg Leu Ala Leu Ile Ile Cys Asn Thr Lys Phe Asp His Leu Pro Ala
180 185 190
Arg Asn Gly Ala His Tyr Asp Ile Val Gly Met Lys Arg Leu Leu Gln
195 200 205
Gly Leu Gly Tyr Thr Val Val Asp Glu Lys Asn Leu Thr Ala Arg Asp
210 215 220
Met Glu Ser Val Leu Arg Ala Phe Ala Ala Arg Pro Glu His Lys Ser
225 230 235 240
Ser Asp Ser Thr Phe Leu Val Leu Met Ser His Gly Ile Leu Glu Gly
245 250 255
Ile Cys Gly Thr Ala His Lys Lys Lys Lys Pro Asp Val Leu Leu Tyr
260 265 270
Asp Thr Ile Phe Gln Ile Phe Asn Asn Arg Asn Cys Leu Ser Leu Lys
275 280 285
Asp Lys Pro Lys Val Ile Ile Val Gln Ala Cys Arg Gly Glu Lys His
290 295 300
Gly Glu Leu Trp ValArg Asp Ser Pro Ala Ser Leu Ala Val Ile Ser
305 310 315 320
Ser Gln Ser Ser Glu Asn Leu Glu Ala Asp Ser Val Cys Lys Ile His
325 330 335
Glu Glu Lys Asp Phe Ile Ala Phe Cys Ser Ser Thr Pro His Asn Val
340 345 350
Ser Trp Arg Asp Arg Thr Arg Gly Ser Ile Phe Ile Thr Glu Leu Ile
355 360 365
Thr Cys Phe Gln Lys Tyr Ser Cys Cys Cys His Leu Met Glu Ile Phe
370 375 380
Arg Lys Val Gln Lys Ser Phe Glu Val Pro Gln Ala Lys Ala Gln Met
385 390 395 400
Pro Thr Ile Glu Arg Ala Thr Leu Thr Arg Asp Phe Tyr Leu Phe Pro
405 410 415
Gly Asn
<210>348
<211>1400
<212>DNA
<213>Homo sapiens
<220>
<223>Caspase-5 precursor C14.008
<400>348
cggcaaaaaa aaaaggcgta agaattttga agctatgttc aaaggtatcc ttcagagtgg 60
attggataac ttcgtgataa accacatgct aaagaacaac gtggctggac aaacatctat 120
ccagacccta gtacctaata cggatcaaaa gtcgaccagt gtaaaaaaag acaaccacaa 180
aaaaaaaaca gttaagatgt tggaatacct gggcaaagat gttcttcatg gtgtttttaa 240
ttatttggca aaacacgatg ttctgacatt gaaggaagag gaaaagaaaa aatattatga 300
tgccaaaatt gaagacaagg ccctgatctt ggtagactct ttgcgaaaga atcgcgtggc 360
tcatcaaatg tttacccaaa cacttctcaa tatggaccaa aagatcacca gtgtaaaacc 420
tcttctgcaa atcgaggctg gaccacctga gtcagcagaa tctacaaata tactcaaact 480
ttgtcctcgt gaagaattcc tgagactgtg taaaaaaaat catgatgaga tctatccaat 540
aaaaaagaga gaggaccgca gacgcctggc tctcatcata tgcaatacaa agtttgatca 600
cctgcctgca aggaatgggg ctcactatga catcgtgggg atgaaaaggc tgcttcaagg 660
cctgggctac actgtggttg acgaaaagaa tctcacagcc agggatatgg agtcagtgct 720
gagggcattt gctgccagac cagagcacaa gtcctctgac agcacgttct tggtactcat 780
gtctcatggc atcctagagg gaatctgcgg aactgcgcat aaaaagaaaa aaccggatgt 840
gctgctttat gacaccatct tccagatatt caacaaccgc aactgcctca gtctaaagga 900
caaacccaag gtcatcattg tccaggcctg cagaggtgaa aaacatgggg aactctgggt 960
cagagactct ccagcatcct tggcagtcat ctcttcacag tcatctgaga acctggaggc 1020
agattctgtt tgcaagatcc acgaggagaa ggacttcatt gctttctgtt cttcaacacc 1080
acataacgtg tcctggagag accgcacaag gggctccatc ttcattacgg aactcatcac 1140
atgcttccag aaatattctt gctgctgcca cctaatggaa atatttcgga aggtacagaa 1200
atcatttgaa gttccacagg ctaaagccca gatgcccacc atagaacgag caaccttgac 1260
aagagatttc tacctctttc ctggcaattg aaaatgaaac cacaggcagc ccagccctcc 1320
tctgtcaaca tcaaagagca catttaccag tatagcttgc atagtcaata tttggtattt 1380
caataaaagt aaagactgta 1400
<210>349
<211>479
<212>PRT
<213>Homo sapiens
<220>
<223>Caspase-8 precursor C14.009
<400>349
Met Asp Phe Ser Arg Asn Leu Tyr Asp Ile Gly Glu Gln Leu Asp Ser
1 5 10 15
Glu Asp Leu Ala Ser Leu Lys Phe Leu Ser Leu Asp Tyr Ile Pro Gln
20 25 30
Arg Lys Gln Glu Pro Ile Lys Asp Ala Leu Met Leu Phe Gln Arg Leu
35 40 45
Gln Glu Lys Arg Met Leu Glu Glu Ser Asn Leu Ser Phe Leu Lys Glu
50 55 60
Leu Leu Phe Arg Ile Asn Arg Leu Asp Leu Leu Ile Thr Tyr Leu Asn
65 70 75 80
Thr Arg Lys Glu Glu Met Glu Arg Glu Leu Gln Thr Pro Gly Arg Ala
85 90 95
Gln Ile Ser Ala Tyr Arg Val Met Leu Tyr Gln Ile Ser Glu Glu Val
100 105 110
Ser Arg Ser Glu Leu Arg Ser Phe Lys Phe Leu Leu Gln Glu Glu Ile
115 120 125
Ser Lys Cys Lys Leu Asp Asp Asp Met Asn Leu Leu Asp Ile Phe Ile
130 135 140
Glu Met Glu Lys Arg Val Ile Leu Gly Glu Gly Lys Leu Asp Ile Leu
145 150 155 160
Lys Arg Val Cys Ala Gln Ile Asn Lys Ser Leu Leu Lys Ile Ile Asn
165 170 175
Asp Tyr Glu Glu Phe Ser Lys Glu Arg Ser Ser Ser Leu Glu Gly Ser
180 185 190
Pro Asp Glu Phe Ser Asn Gly Glu Glu Leu Cys Gly Val Met Thr Ile
195 200 205
Ser Asp Ser Pro Arg Glu Gln Asp Ser Glu Ser Gln Thr Leu Asp Lys
210 215 220
Val Tyr Gln Met Lys Ser Lys Pro Arg Gly Tyr Cys Leu Ile Ile Asn
225 230 235 240
Asn His Asn Phe Ala Lys Ala Arg Glu Lys Val Pro Lys Leu His Ser
245 250 255
Ile Arg Asp Arg Asn Gly Thr His Leu Asp Ala Gly Ala Leu Thr Thr
260 265 270
Thr Phe Glu Glu Leu His Phe Glu Ile Lys Pro His Asp Asp Cys Thr
275 280 285
Val Glu Gln Ile Tyr Glu Ile Leu Lys Ile Tyr Gln Leu Met Asp His
290 295 300
Ser Asn Met Asp Cys Phe Ile Cys Cys Ile Leu Ser His Gly Asp Lys
305 310 315 320
Gly Ile Ile Tyr Gly Thr Asp Gly Gln Glu Ala Pro Ile Tyr Glu Leu
325 330 335
Thr Ser Gln Phe Thr Gly Leu Lys Cys Pro Ser Leu Ala Gly Lys Pro
340 345 350
Lys Val Phe Phe Ile Gln Ala Cys Gln Gly Asp Asn Tyr Gln Lys Gly
355 360 365
Ile Pro Val Glu Thr Asp Ser Glu Glu Gln Pro Tyr Leu Glu Met Asp
370 375 380
Leu Ser Ser Pro Gln Thr Arg Tyr Ile Pro Asp Glu Ala Asp Phe Leu
385 390 395 400
Leu Gly Met Ala Thr Val Asn Asn Cys Val Ser Tyr Arg Asn Pro Ala
405 410 415
Glu Gly Thr Trp Tyr Ile Gln Ser Leu Cys Gln Ser Leu Arg Glu Arg
420 425 430
Cys Pro Arg Gly Asp Asp Ile Leu Thr Ile Leu Thr Glu Val Asn Tyr
435 440 445
Glu Val Ser Asn Lys Asp Asp Lys Lys Asn Met Gly Lys Gln Met Pro
450 455 460
Gln Pro Thr Phe Thr Leu Arg Lys Lys Leu Val Phe Pro Ser Asp
465 470 475
<210>350
<211>2887
<212>DNA
<213>Homo sapiens
<220>
<223>Caspase-8 precursor C14.009
<400>350
gattctgcct ttctgctgga gggaagtgtt ttcacaggtt ctcctccttt tatcttttgt 60
gttttttttc aagccctgct gaatttgcta gtcaactcaa caggaagtga ggccatggag 120
ggaggcagaa gagccagggt ggttattgaa agtagaagaa acttcttcct gggagccttt 180
cccaccccct tccctgctga gcacgtggag ttaggcaggt taggggactc ggagactgcg 240
atggtgccag gaaagggtgg agcggattat attctcctgc cttttaaaaa gatggacttc 300
agcagaaatc tttatgatat tggggaacaa ctggacagtg aagatctggc ctccctcaag 360
ttcctgagcc tggactacat tccgcaaagg aagcaagaac ccatcaagga tgccttgatg 420
ttattccaga gactccagga aaagagaatg ttggaggaaa gcaatctgtc cttcctgaag 480
gagctgctct tccgaattaa tagactggat ttgctgatta cctacctaaa cactagaaag 540
gaggagatgg aaagggaact tcagacacca ggcagggctc aaatttctgc ctacagggtc 600
atgctctatc agatttcaga agaagtgagc agatcagaat tgaggtcttt taagtttctt 660
ttgcaagagg aaatctccaa atgcaaactg gatgatgaca tgaacctgct ggatattttc 720
atagagatgg agaagagggt catcctggga gaaggaaagt tggacatcct gaaaagagtc 780
tgtgcccaaa tcaacaagag cctgctgaag ataatcaacg actatgaaga attcagcaaa 840
gagagaagca gcagccttga aggaagtcct gatgaatttt caaatgggga ggagttgtgt 900
ggggtaatga caatctcgga ctctccaaga gaacaggata gtgaatcaca gactttggac 960
aaagtttacc aaatgaaaag caaacctcgg ggatactgtc tgatcatcaa caatcacaat 1020
tttgcaaaag cacgggagaa agtgcccaaa cttcacagca ttagggacag gaatggaaca 1080
cacttggatg caggggcttt gaccacgacc tttgaagagc ttcattttga gatcaagccc 1140
cacgatgact gcacagtaga gcaaatctat gagattttga aaatctacca actcatggac 1200
cacagtaaca tggactgctt catctgctgt atcctctccc atggagacaa aggcatcatc 1260
tatggcactg atggacagga ggcccccatc tatgagctga catctcagtt cactggtttg 1320
aagtgccctt cccttgctgg aaaacccaaa gtgtttttta ttcaggcttg tcagggggat 1380
aactaccaga aaggtatacc tgttgagact gattcagagg agcaacccta tttagaaatg 1440
gatttatcat cacctcaaac gagatatatc ccggatgagg ctgactttct gctggggatg 1500
gccactgtga ataactgtgt ttcctaccga aaccctgcag agggaacctg gtacatccag 1560
tcactttgcc agagcctgag agagcgatgt cctcgaggcg atgatattct caccatcctg 1620
actgaagtga actatgaagt aagcaacaag gatgacaaga aaaacatggg gaaacagatg 1680
cctcagccta ctttcacact aagaaaaaaa cttgtcttcc cttctgattg atggtgctat 1740
tttgtttgtt ttgttttgtt ttgttttttt gagacagaat ctcgctctgt cgcccaggct 1800
ggagtgcagt ggcgtgatct cggctcaccg caagctccgc ctcccgggtt cacgccattc 1860
tcctgcctca gcctcccgag tagctgggac tacaggggcc cgccaccaca cctggctaat 1920
tttttaaaaa tatttttagt agagacaggg tttcactgtg ttagccaggg tggtcttgat 1980
ctcctgacct cgtgatccac ccacctcggc ctcccaaagt gctgggatta caggcgtgag 2040
ccaccgcgcc tggccgatgg tactatttag atataacact atgtttattt actaattttc 2100
tagattttct actttattaa ttgttttgca cttttttata agagctaaag ttaaatagga 2160
tattaacaac aataacactg tctcctttct cttacgctta aggctttggg aatgttttta 2220
gctggtggca ataaatacca gacacgtaca aaatccagct atgaatatag agggcttatg 2280
attcagattg ttatctatca actataagcc cactgttaat attctattaa ctttaattct 2340
ctttcaaagc taaattccac actaccacat taaaaaaatt agaaagtagc cacgtatggt 2400
ggctcatgtc tataatccca gcactttggg aggttgaggt gggaggattt gcttgaaccc 2460
aagaggtcca aggctgcagt gagccatgtt cacaccgctg cactcaagct tgggtgacag 2520
agcaagaccc cgtccccaaa aaaatttttt ttttaataaa cccaaatttg tttgaaaact 2580
tttaaaaatt caaatgattt ttacaagttt taaataagct ctccccaaac ttgctttatg 2640
ccttcttatt gcttttatga tatatatatg cttggctaac tatatttgct ttttgctaac 2700
aatgctctgg ggtcttttta tgcatttgca tttgctcttt catctctgct tggattattt 2760
taaatcatta ggaattaagt tatctttaaa atttaagtat cttttttcca aaacattttt 2820
taatagaata aaatataatt tgatcttaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2880
aaaaaaa 2887
<210>351
<211>416
<212>PRT
<213>Homo sapiens
<220>
<223>Caspase-9 precursor
C14.010
<400>351
Met Asp Glu Ala Asp Arg Arg Leu Leu Arg Arg Cys Arg Leu Arg Leu
1 5 10 15
Val Glu Glu Leu Gln Val Asp Gln Leu Trp Asp Ala Leu Leu Ser Arg
20 25 30
Glu Leu Phe Arg Pro His Met Ile Glu Asp Ile Gln Arg Ala Gly Ser
35 40 45
Gly Ser Arg Arg Asp Gln Ala Arg Gln Leu Ile Ile Asp Leu Glu Thr
50 55 60
Arg Gly Ser Gln Ala Leu Pro Leu Phe Ile Ser Cys Leu Glu Asp Thr
65 70 75 80
Gly Gln Asp Met Leu Ala Ser Phe Leu Arg Thr Asn Arg Gln Ala Ala
85 90 95
Lys Leu Ser Lys Pro Thr Leu Glu Asn Leu Thr Pro Val Val Leu Arg
100 105 110
Pro Glu Ile Arg Lys Pro Glu Val Leu Arg Pro Glu Thr Pro Arg Pro
115 120 125
Val Asp Ile Gly Ser Gly Gly Phe Gly Asp Val Gly Ala Leu Glu Ser
130 135 140
Leu Arg Gly Asn Ala Asp Leu Ala Tyr Ile Leu Ser Met Glu Pro Cys
145 150 155 160
Gly His Cys Leu Ile Ile Asn Asn Val Asn Phe Cys Arg Glu Ser Gly
165 170 175
Leu Arg Thr Arg Thr Gly Ser Asn Ile Asp Cys Glu Lys Leu Arg Arg
180 185 190
Arg Phe Ser Ser Leu His Phe Met Val Glu Val Lys Gly Asp Leu Thr
195 200 205
Ala Lys Lys Met Val Leu Ala Leu Leu Glu Leu Ala Gln Gln Asp His
210 215 220
Gly Ala Leu Asp Cys Cys Val Val Val Ile Leu Ser His Gly Cys Gln
225 230 235 240
Ala Ser His Leu Gln Phe Pro Gly Ala Val Tyr Gly Thr Asp Gly Cys
245 250 255
Pro Val Ser Val Glu Lys Ile Val Asn Ile Phe Asn Gly Thr Ser Cys
260 265 270
Pro Ser Leu Gly Gly Lys Pro Lys Leu Phe Phe Ile Gln Ala Cys Gly
275 280 285
Gly Glu Gln Lys Asp His Gly Phe Glu Val Ala Ser Thr Ser Pro Glu
290 295 300
Asp Glu Ser Pro Gly Ser Asn Pro Glu Pro Asp Ala Thr Pro Phe Gln
305 310 315 320
Glu Gly Leu Arg Thr Phe Asp Gln Leu Asp Ala Ile Ser Ser Leu Pro
325 330 335
Thr Pro Ser Asp Ile Phe Val Ser Tyr Ser Thr Phe Pro Gly Phe Val
340 345 350
Ser Trp Arg Asp Pro Lys Ser Gly Ser Trp Tyr Val Glu Thr Leu Asp
355 360 365
Asp Ile Phe Glu Gln Trp Ala His Ser Glu Asp Leu Gln Ser Leu Leu
370 375 380
Leu Arg Val Ala Asn Ala Val Ser Val Lys Gly Ile Tyr Lys Gln Met
385 390 395 400
Pro Gly Cys Phe Asn Phe Leu Arg Lys Lys Leu Phe Phe Lys Thr Ser
405 410 415
<210>352
<211>1481
<212>DNA
<213>Homo sapiens
<220>
<223>Caspase-9 precursor C14.010
<400>352
gccatggacg aagcggatcg gcggctcctg cggcggtgcc ggctgcggct ggtggaagag 60
ctgcaggtgg accagctctg ggacgtcctg ctgagccgcg agctgttcag gccccatatg 120
atcgaggaca tccagcgggc aggctctgga tctcggcggg atcaggccag gcagctgatc 180
atagatctgg agactcgagg gagtcaggct cttcctttgt tcatctcctg cttagaggac 240
acaggccagg acatgctggc ttcgtttctg cgaactaaca ggcaagcagg aaagttgtcg 300
aagccaaccc tagaaaacct taccccagtg gtgctcagac cagagattcg caaaccagag 360
gttctcagac cggaaacacc cagaccagtg gacattggtt ctggaggatt cggtgatgtc 420
ggtgctcttg agagtttgag gggaaatgca gatttggctt acatcctgag catggagccc 480
tgtggccact gcctcattat caacaatgtg aacttctgcc gtgagtccgg gctccgcacc 540
cgcactggct ccaacatcga ctgtgagaag ttgcggcgtc gcttctcctc gctgcatttc 600
atggtggagg tgaagggcga cctgactgcc aagaaaatgg tgctggcttt gctggagctg 660
gcgcggcagg accacggtgc tctggactgc tgcgtggtgg tcattctctc tcacggctgt 720
caggccagcc acctgcagtt cccaggggct gtctacggca cagatggatg ccctgtgtcg 780
gtcgagaaga ttgtgaacat cttcaatggg accagctgcc ccagcctggg agggaagccc 840
aagctctttt tcatccaggc ctgtggtggg gagcagaaag accatgggtt tgaggtggcc 900
tccacttccc ctgaagacga gtcccctggc agtaaccccg agccagatgc caccccgttc 960
caggaaggtt tgaggacctt cgaccagctg gacgccatat ctagtttgcc cacacccagt 1020
gacatctttg tgtcctactc tactttccca ggttttgttt cctggaggga ccccaagagt 1080
ggctcctggt acgttgagac cctggacgac atctttgagc agtgggctca ctctgaagac 1140
ctgcagtccc tcctgcttag ggtcgctaat gctgtttcgg tgaaagggat ttataaacag 1200
atgcctggtt gctttaattt cctccggaaa aaacttttct ttaaaacatc ataaggccag 1260
ggcccctcac cctgccttat cttgcacccc caaagctttc ctgccccagg cctgaaagag 1320
gctgaggcct ggactttcct gcaactcaag gactttgcag ccggcacagg gtctgctctt 1380
tctctgccag tgacagacag gctcttagca gcttccagat tgacgacaag tgctgaacag 1440
tggaggaaga gggacagatg aatgccgtgg attgcacgtg g 1481
<210>353
<211>521
<212>PRT
<213>Homo sapiens
<220>
<223>Caspase-l0 precursor C14.011
<400>353
Met Lys Ser Gln Gly Gln His Trp Tyr Ser Ser Ser Asp Lys Asn Cys
1 5 10 15
Lys Val Ser Phe Arg Glu Lys Leu Leu Ile Ile Asp Ser Asn Leu Gly
20 25 30
Val Gln Asp Val Glu Asn Leu Lys Phe Leu Cys Ile Gly Leu Val Pro
35 40 45
Asn Lys Lys Leu Glu Lys Ser Ser Ser Ala Ser Asp Val Phe Glu His
50 55 60
Leu Leu Ala Glu Asp Leu Leu Ser Glu Glu Asp Pro Phe Phe Leu Ala
65 70 75 80
Glu Leu Leu Tyr Ile Ile Arg Gln Lys Lys Leu Leu Gln His Leu Asn
85 90 95
Cys Thr Lys Glu Glu Val Glu Arg Leu Leu Pro Thr Arg Gln Arg Val
100 105 110
Ser Leu Phe Arg Asn Leu Leu Tyr Glu Leu Ser Glu Gly Ile Asp Ser
115 120 125
Glu Asn Leu Lys Asp Met Ile Phe Leu Leu Lys Asp Ser Leu Pro Lys
130 135 140
Thr Glu Met Thr Ser Leu Ser Phe Leu Ala Phe Leu Glu Lys Gln Gly
145 150 155 160
Lys Ile Asp Glu Asp Asn Leu Thr Cys Leu Glu Asp Leu Cys Lys Thr
165 170 175
Val Val Pro Lys Leu Leu Arg Asn Ile Glu Lys Tyr Lys Arg Glu Lys
180 185 190
Ala Ile Gln Ile Val Thr Pro Pro Val Asp Lys Glu Ala Glu Ser Tyr
195 200 205
Gln Gly Glu Glu Glu Leu Val Ser Gln Thr Asp Val Lys Thr Phe Leu
210 215 220
Glu Ala Leu Pro Gln Glu Ser Trp Gln Asn Lys His Ala Gly Ser Asn
225 230 235 240
Gly Asn Arg Ala Thr Asn Gly Ala Pro Ser Leu Val Ser Arg Gly Met
245 250 255
Gln Gly Ala Ser Ala Asn Thr Leu Asn Ser Glu Thr Ser Thr Lys Arg
260 265 270
Ala Ala Val Tyr Arg Met Asn Arg Asn His Arg Gly Leu Cys Val Ile
275 280 285
Val Asn Asn His Ser Phe Thr Ser Leu Lys Asp Arg Gln Gly Thr His
290 295 300
Lys Asp Ala Glu Ile Leu Ser His Val Phe Gln Trp Leu Gly Phe Thr
305 310 315 320
Val His Ile His Asn Asn Val Thr Lys Val Glu Met Glu Met Val Leu
325 330 335
Gln Lys Gln Lys Cys Asn Pro Ala His Ala Asp Gly Asp Cys Phe Val
340 345 350
Phe Cys Ile Leu Thr His Gly Arg Phe Gly Ala Val Tyr Ser Ser Asp
355 360 365
Glu Ala Leu Ile Pro Ile Arg Glu Ile Met Ser His Phe Thr Ala Leu
370 375 380
Gln Cys Pro Arg Leu Ala Glu Lys Pro Lys Leu Phe Phe Ile Gln Ala
385 390 395 400
Cys Gln Gly Glu Glu Ile Gln Pro Ser Val Ser Ile Glu Ala Asp Ala
405 410 415
Leu Asn Pro Glu Gln Ala Pro Thr Ser Leu Gln Asp Ser Ile Pro Ala
420 425 430
Glu Ala Asp Phe Leu Leu Gly Leu Ala Thr Val Pro Gly Tyr Val Ser
435 440 445
Phe Arg His Val Glu Glu Gly Ser Trp Tyr Ile Gln Ser Leu Cys Asn
450 455 460
His Leu Lys Lys Leu Val Pro Arg Met Leu Lys Phe Leu Glu Lys Thr
465 470 475 480
Met Glu Ile Arg Gly Arg Lys Arg Thr Val Trp Gly Ala Lys Gln Ile
485 490 495
Ser Ala Thr Ser Leu Pro Thr Ala Ile Ser Ala Gln Thr Pro Arg Pro
500 505 510
Pro Met Arg Arg Trp Ser Ser Val Ser
515 520
<210>354
<211>3536
<212>DNA
<213>Homo sapiens
<220>
<223>Caspase-10 precursor C14.011
<400>354
tgaagtctct tcccaagcaa atgggagctt ctttggacct tggagcacac agaggattct 60
actttcttta aaactttgtt ttcaggcaat ttccctgaga accgtttact tccagaagat 120
tggtggagct tgatctgaag gctggccatg aaatctcaag gtcaacattg gtattccagt 180
tcagataaaa actgtaaagt gagctttcgt gagaagcttc tgattattga ttcaaacctg 240
ggggtccaag atgtggagaa cctcaagttt ctctgcatag gattggtccc caacaagaag 300
ctggagaagt ccagctcagc ctcagatgtt tttgaacatc tcttggcaga ggatctgctg 360
agtgaggaag accctttctt cctggcagaa ctcctctata tcatacggca gaagaagctg 420
ctgcagcacc tcaactgtac caaagaggaa gtggagcgac tgctgcccac ccgacaaagg 480
gtttctctgt ttagaaacct gctctacgaa ctgtcagaag gcattgactc agagaactta 540
aaggacatga tcttccttct gaaagactcg cttcccaaaa ctgaaatgac ctccctaagt 600
ttcctggcat ttctagagaa acaaggtaaa atagatgaag ataatctgac atgcctggag 660
gacctctgca aaacagttgt acctaaactt ttgagaaaca tagagaaata caaaagagag 720
aaagctatcc agatagtgac acctcctgta gacaaggaag ccgagtcgta tcaaggagag 780
gaagaactag tttcccaaac agatgttaag acattcttgg aagccttacc gagggcagct 840
gtgtacagga tgaatcggaa ccacagaggc ctctgtgtca ttgtcaacaa ccacagcttt 900
acctccctga aggacagaca aggaacccat aaagatgctg agatcctgag tcatgtgttc 960
cagtggcttg ggttcacagt gcatatacac aataatgtga cgaaagtgga aatggagatg 1020
gtcctgcaga agcagaagtg caatccagcc catgccgacg gggactgctt cgtgttctgt 1080
attctgaccc atgggagatt tggagctgtc tactcttcgg atgaggccct cattcccatt 1140
cgggagatca tgtctcactt cacagccctg cagtgcccta gactggctga aaaacctaaa 1200
ctctttttca tccaggcctg ccaaggtgaa gagatacagc cttccgtatc catcgaagca 1260
gatgctctga accctgagca ggcacccact tccctgcagg acagtattcc tgccgaggct 1320
gacttcctac ttggtctggc cactgtccca ggctatgtat cctttcggca tgtggaggaa 1380
ggcagctggt atattcagtc tctgtgtaat catctgaaga aattggtccc aagacatgaa 1440
gacatcttat ccatcctcac tgctgtcaac gatgatgtga gtcgaagagt ggacaaacag 1500
ggaacaaaga aacagatgcc ccagcctgct ttcacactaa ggaaaaaact agtattccct 1560
gtgcccctgg atgcactttc aatatagcag agagtttttg ttggttctta gacctcaaac 1620
gaatcattgg gtataacctc cagcctcctg cccagcacag gaatcggtgg tctccacctg 1680
tcattctaga aacaggaaac accgtgtttt ctgacacagt caattctgat tttctttttc 1740
ttttgcaagt ctaaatgtta gaaaactttc tttttttgga gatagtctca ttctgtcacc 1800
cagactggag tgcagggggg caatcacggc tcactgtagt ctcgacctcc caggctcaag 1860
ctgtcctccc acctcagcct cccaagtagc tgagactaca ggtgtgtgtc catgcacagc 1920
taacttttta ttttttttgt ggagatgggg tttcactatg ttgcctaagc tggtctcaaa 1980
ctcctgggct caagcgatcc tcccacctca gcttctcaaa gttctgggac tacaggcatg 2040
aaatactgtg cctggcctgg ggaccaggtg cattttaagg ttccttggtg ttcaaaaacc 2100
acgttcttag cctagattga gcttagattg cctctctaga caactacccc ttagttataa 2160
ttctgtgtcc cctctgcatg cccttaaaca ttggacagtg aggtcacagt ccacccaccc 2220
tctctctgat ctcccccttc ctaagacttc tcttttgcac atctagtgag gtgaaaattt 2280
ggtctatgcc aggcccattt cctgcttttg tgtaaggaag gtgctcacat aggaagtttt 2340
tatttggtta gagacaggtt tccctgtagg aagatgatgg ctcatttaca ctcagctgct 2400
ctgcaagcag aaactttaca acctgatgtc atattccatt ttggactggg tgcggtgact 2460
catgcctgta atcccagtac tctgggaagc caaggcaggc agatcacttg aggtcaggag 2520
ttcgagacca gcctggccaa tacggcaaaa cctcatcatt actaaaaaca caaaaattag 2580
ccaggtgtgg cggcgagcac ctgtaatccc agctactcgg gaggctgaga caggagaatc 2640
tcttgaatcc aggaggcaga ggctgtggtg agccaagatg acacaactgc actccagctt 2700
gggcaacagg gcgagacctt gtttaaaaaa aaaattcaat attggggttg gaacatttca 2760
gttgccattg acagaacacc caattcaaat tgactgaagc aaagaaggga atttattgcc 2820
tctttcacat tgaaacccag gagtggataa cactggcttc aggcaaagct tgaatcagga 2880
ctcaatctac aggccagcac ctttctcttg gccggatgtc ctcagggctg gcagatgcag 2940
tagactgcag tggacagtcc ccaccttgtt actgctacta cactttgctc ctctggccca 3000
aggcatgagg agagaggctg tgtcagaaac tgaagctgtt ctcaggatca ctgggctctt 3060
cttggcagag gggatgtctg gcttgcctga agggagtggc tctgtaagga cgccttgatg 3120
ctttcttcat taagattttg agcattttta cgtacttgag cttttttttt tttttttttc 3180
aatttctaga ggaacttttt ctctgttaat tcctggaact gtattttgaa tccttaaagg 3240
tgagccctca tagggagatc caaagtcctg tggttaacgc cttcatttat agatgaggca 3300
gctgaggcct ggggatgtga acaacctgct cacagtcctc atttactgga tttgacttca 3360
gccaggtgaa ctggaatgcc ttggggcgtg gaagggcatt aggagtgttt catttgatat 3420
gtgaatgctc ataaaaaaat gtcaaggaat gaagaacaac aactctcagt ggtgcctgca 3480
tttataatta tttatgtgaa agtcaaattc atgtacagta aatttgttat aagaat 3536
<210>355
<211>373
<212>PRT
<213>Mus musculus
<220>
<223>Caspase-11 C14.012
<400>355
Met Ala Glu Asn Lys His Pro Asp Lys Pro Leu Lys Val Leu Glu Gln
1 5 10 15
Leu Gly Lys Glu Val Leu Thr Glu Tyr Leu Glu Lys Leu Val Gln Ser
20 25 30
Asn Val Leu Lys Leu Lys Glu Glu Asp Lys Gln Lys Phe Asn Asn Ala
35 40 45
Glu Arg Ser Asp Lys Arg Trp Val Phe Val Asp Ala Met Lys Lys Lys
50 55 60
His Ser Lys Val Gly Glu Met Leu Leu Gln Thr Phe Phe Ser Val Asp
65 70 75 80
Pro Gly Ser His His Gly Glu Ala Asn Leu Glu Met Glu Glu Pro Glu
85 90 95
Glu Ser Leu Asn Thr Leu Lys Leu Cys Ser Pro Glu Glu Phe Thr Arg
100 105 110
Leu Cys Arg Glu Lys Thr Gln Glu Ile Tyr Pro Ile Lys Glu Ala Asn
115 120 125
Gly Arg Thr Arg Lys Ala Leu Ile Ile Cys Asn Thr Glu Phe Lys His
130 135 140
Leu Ser Leu Arg Tyr Gly Ala Lys Phe Asp Ile Ile Gly Met Lys Gly
145 150 155 160
Leu Leu Glu Asp Leu Gly Tyr Asp Val Val Val Lys Glu Glu Leu Thr
165 170 175
Ala Glu Gly Met Glu Ser Glu Met Lys Asp Phe Ala Ala Leu Ser Glu
180 185 190
His Gln Thr Ser Asp Ser Thr Phe Leu Val Leu Met Ser His Gly Thr
195 200 205
Leu His Gly Ile Cys Gly Thr Met His Ser Glu Lys Thr Pro Asp Val
210 215 220
Leu Gln Tyr Asp Thr Ile Tyr Gln Ile Phe Asn Asn Cys His Cys Pro
225 230 235 240
Gly Leu Arg Asp Lys Pro Lys Val Ile Ile Val Gln Ala Cys Arg Gly
245 250 255
Gly Asn Ser Gly Glu Met Trp Ile Arg Glu Ser Ser Lys Pro Gln Leu
260 265 270
Cys Arg Gly Val Asp Leu Pro Arg Asn Met Glu Ala Asp Ala Val Lys
275 280 285
Leu Ser His Val Glu Lys Asp Phe Ile Ala Phe Tyr Ser Thr Thr Pro
290 295 300
His His Leu Ser Tyr Arg Asp Lys Thr Gly Gly Ser Tyr Phe Ile Thr
305 310 315 320
Arg Leu Ile Ser Cys Phe Arg Lys His Ala Cys Ser Cys His Leu Phe
325 330 335
Asp Ile Phe Leu Lys Val Gln Gln Ser Phe Glu Lys Ala Ser Ile His
340 345 350
Ser Gln Met Pro Thr Ile Asp Arg Ala Thr Leu Thr Arg Tyr Phe Tyr
355 360 365
Leu Phe Pro Gly Asn
370
<210>356
<211>1350
<212>DNA
<213>Mus musculus
<220>
<223>Caspase-11 C14.012
<400>356
tcttcacagt gcgaaagaac tgaggctttt tctcatggct gaaaacaaac accctgacaa 60
accacttaag gtgttggaac agctgggcaa agaagtcctt acggagtacc tagaaaaatt 120
agtacaaagc aatgtactga aattaaagga ggaagataaa caaaaattta acaatgctga 180
acgcagtgac aagcgttggg tttttgtaga tgccatgaaa aagaaacaca gcaaagtagg 240
tgaaatgctt ctccagacat tcttcagtgt ggacccaggc agccaccatg gtgaagctaa 300
tctggaaatg gaggaaccag aagaatcatt gaacactctc aagctttgtt cccctgaaga 360
gttcacaagg ctttgcagag aaaagacaca agaaatttac ccaataaagg aggccaatgg 420
ccgtacacga aaggctctta tcatatgcaa tacagagttc aaacatctct cactgaggta 480
tggggctaaa tttgacatca ttggtatgaa aggccttctt gaagacttag gctacgatgt 540
ggtggtgaaa gaggagctta cagcagaggg catggagtca gagatgaaag actttgctgc 600
actctcagaa caccagacat cagacagcac attcctggtg ctaatgtctc atggcacact 660
gcatggcatt tgtggaacaa tgcacagtga aaaaactcca gatgtgctac agtatgatac 720
catctatcag atattcaaca attgccactg tccaggtcta cgagacaaac ccaaagtcat 780
cattgtgcag gcctgcagag gtgggaactc tggagaaatg tggatcagag agtcttcaaa 840
accccagttg tgcagaggtg tagatctacc taggaatatg gaagctgatg ctgtcaagct 900
gagccacgtg gagaaggact tcattgcctt ctactctaca accccacatc acttgtccta 960
ccgagacaaa acaggaggct cttacttcat cactagactc atttcctgct tccggaaaca 1020
tgcttgctct tgtcatctct ttgatatatt cctgaaggtg caacaatcat ttgaaaaggc 1080
aagtattcat tcccagatgc ccaccattga tcgggcaacc ttgacaagat atttctacct 1140
ctttcctggc aactgagaac aaagcaacaa gcaactgaat ctcatttctt cagcttgaag 1200
aagtgatctt ggccaaggat cacattctat tcctgaaatt ccagaactag tgaaattaag 1260
gaaagaatac ttatgaattc aagaccagcc taagcaacac agtgggattc tgttccatag 1320
acaagcaaac aagcaaaaat aaaaaaaaaa 1350
<210>357
<211>419
<212>PRT
<213>Mus musculus
<220>
<223>Caspase-12 precursor
C14.013
<400>357
Met Ala Ala Arg Arg Thr His Glu Arg Asp Pro Ile Tyr Lys Ile Lys
1 5 10 15
Gly Leu Ala Lys Asp Met Leu Asp Gly Val Phe Asp Asp Leu Val Glu
20 25 30
Lys Asn Val Leu Asn Gly Asp Glu Leu Leu Lys Ile Gly Glu Ser Ala
35 40 45
Ser Phe Ile Leu Asn Lys Ala Glu Asn Leu Val Glu Asn Phe Leu Glu
50 55 60
Lys Thr Asp Met Ala Gly Lys Ile Phe Ala Gly His Ile Ala Asn Ser
65 70 75 80
Gln Glu Gln Leu Ser Leu Gln Phe Ser Asn Asp Glu Asp Asp Gly Pro
85 90 95
Gln Lys Ile Cys Thr Pro Ser Ser Pro Ser Glu Ser Lys Arg Lys Val
100 105 110
Glu Asp Asp Glu Met Glu Val Asn Ala Gly Leu Ala His Glu Ser His
115 120 125
Leu Met Leu Thr Ala Pro His Gly Leu Gln Ser Ser Glu Val Gln Asp
130 135 140
Thr Leu Lys Leu Cys Pro Arg Asp Gln Phe Cys Lys Ile Lys Thr Glu
145 150 155 160
Arg Ala Lys Glu Ile Tyr Pro Val Met Glu Lys Glu Gly Arg Thr Arg
165 170 175
Leu Ala Leu Ile Ile Cys Asn Lys Lys Phe Asp Tyr Leu Phe Asp Arg
180 185 190
Asp Asn Ala Asp Thr Asp Ile Leu Asn Met Gln Glu Leu Leu Glu Asn
195 200 205
Leu Gly Tyr Ser Val Val Leu Lys Glu Asn Leu Thr Ala Gln Glu Met
210 215 220
Glu Thr Glu Leu Met Gln Phe Ala Gly Arg Pro Glu His Gln Ser Ser
225 230 235 240
Asp Ser Thr Phe Leu Val Phe Met Ser His Gly Ile Leu Glu Gly Ile
245 250 255
Cys Gly Val Lys His Arg Asn Lys Lys Pro Asp Val Leu His Asp Asp
260 265 270
Thr Ile Phe Lys Ile Phe Asn Asn Ser Asn Cys Arg Ser Leu Arg Asn
275 280 285
Lys Pro Lys Ile Leu Ile Met Gln Ala Cys Arg Gly Arg Tyr Asn Gly
290 295 300
Thr Ile Trp Val Ser Thr Asn Lys Gly Ile Ala Thr Ala Asp Thr Asp
305 310 315 320
Glu Glu Arg Val Leu Ser Cys Lys Trp Asn Asn Ser Ile Thr Lys Ala
325 330 335
His Val Glu Thr Asp Phe Ile Ala Phe Lys Ser Ser Thr Pro His Asn
340 345 350
Ile Ser Trp Lys Val Gly Lys Thr Gly Ser Leu Phe Ile Ser Lys Leu
355 360 365
Ile Asp Cys Phe Lys Lys Tyr Cys Trp Cys Tyr His Leu Glu Glu Ile
370 375 380
Phe Arg Lys Val Gln His Ser Phe Glu Val Pro Gly Glu Leu Thr Gln
385 390 395 400
Met Pro Thr Ile Glu Arg Val Ser Met Thr Arg Tyr Phe Tyr Leu Phe
405 410 415
Pro Gly Asn
<210>358
<211>2228
<212>DNA
<213>Mus musculus
<220>
<223>Caspase-12 precursor C14.013
<400>358
ggcacgagcg gcacgagcgg cacgagcaca gccatggcgg ccaggaggac acatgaaaga 60
gatccaatct acaagatcaa aggtttggcc aaggacatgc tggatggggt ttttgatgac 120
ctggtggaga agaatgtttt aaatggagat gagttactca aaatagggga aagtgcgagt 180
ttcatcctga acaaggctga gaatctggtt gagaacttct tagagaaaac agacatggca 240
ggaaaaatat ttgctggcca cattgccaat tcccaggaac agctgagttt acaattttct 300
aatgatgagg atgatggacc tcagaagata tgtacacctt cttctccatc agaatccaag 360
agaaaagtag aggatgatga aatggaggta aatgctggat tggcccatga atcacatcta 420
atgctgacag ctcctcatgg actccagagc tcagaagtcc aagatacact gaagctttgt 480
ccacgtgatc agttttgtaa gataaagaca gaaagggcaa aagagatata tccagtgatg 540
gagaaggagg gacgaacacg tctggctctc atcatctgca acaaaaagtt tgactacctt 600
tttgatagag ataatgctga tactgacatt ttgaacatgc aagaactact tgaaaatctt 660
ggatactctg tggtgttaaa ggaaaacctt acagctcagg aaatggagac agagttaatg 720
cagtttgctg gccgtccaga gcaccagtcc tcagacagca cattcctggt gtttatgtcc 780
catggcatcc tggaaggaat ctgtggggtg aagcaccgaa acaaaaagcc agatgttctt 840
catgatgaca ctatcttcaa aattttcaac aactctaact gtcggagtct gagaaacaaa 900
cccaagattc tcatcatgca ggcctgcaga ggcagatata atggaactat ttgggtatcc 960
acaaacaaag ggatagccac tgctgataca gatgaggaac gtgtgttgag ctgtaaatgg 1020
aataatagta taacaaaggc ccatgtggag acagatttca ttgctttcaa atcttctacc 1080
ccacataata tttcttggaa ggtaggcaag actggttccc tcttcatttc caaactcatt 1140
gactgcttca aaaagtactg ttggtgttat catttggagg aaatttttcg aaaggttcaa 1200
cactcatttg aggtcccagg tgaactgacc cagatgccca ctattgagag agtatccatg 1260
acacgctatt tctacctttt tcccgggaat tagcacaggc aactctcatg cagttcccag 1320
tcaagtattg ctgtagctga gaagaaaaga aaattccaag atcccaggat ttttaaatgt 1380
gtaaaacttt tcagttagat attttataca atatcataag gctagataat ttccataggt 1440
tatttttagg ggaaacctga tacgtaaaat tttgcatgaa aagtatagat tcgttccttc 1500
cttccaaatc tgctcatctt ttactttatt ttattaaatt tatatttgca catttttgta 1560
aaatgaataa aggaaaaaga cactaaagaa catgtctgaa catatcctgg aacttcttct 1620
ttgtaaaatg aataaaggaa aaagacacta aagaacaaag atgatacacc ttttgatgaa 1680
ccactttcac ctataaatta gtagtaagaa tttactcaca aggaagatct tatctacaat 1740
gtagctgaga aatgaaactg ttgcatatgt aggtaaaact aatcatcttt taggaataag 1800
ataaagaggt ttatgtccac tgtttgtctc tggcatacat ggttttcatg cctagtggct 1860
acctaccatt cctttcaatc aatttcttag gatataattg acaaatttat gctatgttta 1920
taatttttct cttaaatgcc tttgtaataa gatatactca cacatacaca tttaagattc 1980
tgtctattag tgttattgaa ataaatccta gaaacaaaaa ttaaaagttt aaagcattaa 2040
agacaatttt gagacaaaat attctatgaa attctatgca aaattgaatt cttctacaag 2100
tatgtgatga agcccaccta ccactatttt tgtcatcttg aggatgttat atcattattt 2160
ttacttaact gagctcaaat taaaatcttt gtttattctt tgttctataa aaaaaaaaaa 2220
aaaaaaaa 2228
<210>359
<211>339
<212>PRT
<213>Drosophila melanogaster
<220>
<223>Caspase precursor(insect)
C14.015
<400>359
Met Asp Ala Thr Asn Asn Gly Glu Ser Ala Asp Gln Val Gly Ile Arg
1 5 10 15
Val Gly Asn Pro Glu Gln Pro Asn Asp His Thr Asp Ala Leu Gly Ser
20 25 30
Val Gly Ser Gly Gly Ala Gly Ser Ser Gly Leu Val Ala Gly Ser Ser
35 40 45
His Pro Tyr Gly Ser Gly Ala Ile Gly Gln Leu Ala Asn Gly Tyr Ser
50 55 60
Ser Pro Ser Ser Ser Tyr Arg Lys Asn Val Ala Lys Met Val Thr Asp
65 70 75 80
Arg His Ala Ala Glu Tyr Asn Met Arg His Lys Asn Arg Gly Met Ala
85 90 95
Leu Ile Phe Asn His Glu His Phe Glu Val Pro Thr Leu Lys Ser Arg
100 105 110
Ala Gly Thr Asn Val Asp Cys Glu Asn Leu Thr Arg Val Leu Lys Gln
115 120 125
Leu Asp Phe Glu Val Thr Val Tyr Lys Asp Cys Arg Tyr Lys Asp Ile
130 135 140
Leu Arg Thr Ile Glu Tyr Ala Ala Ser Gln Asn His Ser Asp Ser Asp
145 150 155 160
Cys Ile Leu Val Ala Ile Leu Ser His Gly Glu Met Gly Tyr Ile Tyr
165 170 175
Ala Lys Asp Thr Gln Tyr Lys Leu Asp Asn Ile Trp Ser Phe Phe Thr
180 185 190
Ala Asn His Cys Pro Ser Leu Ala Gly Lys Pro Lys Leu Phe Phe Ile
195 200 205
Gln Ala Cys Gln Gly Asp Arg Leu Asp Gly Gly Val Thr Met Gln Arg
210 215 220
Ser Gln Thr Glu Thr Asp Gly Asp Ser Ser Met Ser Tyr Lys Ile Pro
225 230 235 240
Val His Ala Asp Phe Leu Ile Ala Tyr Ser Thr Val Pro Gly Phe Tyr
245 250 255
Ser Trp Arg Asn Thr Thr Arg Gly Ser Trp Phe Met Gln Ser Leu Cys
260 265 270
Ala Glu Leu Ala Ala Asn Gly Lys Arg Leu Asp Ile Leu Thr Leu Leu
275 280 285
Thr Phe Val Cys Gln Arg Val Ala Val Asp Phe Glu Ser Cys Thr Pro
290 295 300
Asp Thr Pro Glu Met His Gln Gln Lys Gln Ile Pro Cys Ile Thr Thr
305 310 315 320
Met Leu Thr Arg Ile Leu Arg Phe Ser Asp Lys Gln Leu Ala Pro Ala
325 330 335
Gly Arg Val
<210>360
<211>1020
<212>DNA
<213>Drosophila melanogaster
<220>
<223>Caspase precursor(insect)C14.015
<400>360
atggacgcca ctaacaatgg agaatccgcc gaccaggtgg gcatcagggt gggcaatccg 60
gagcagccca acgatcacac agatgccctg ggctccgtgg gatccggagg agcgggtagc 120
agcggcctgg tcgcaggatc ctcgcatcct tacggcagcg gagccattgg gcagctggcc 180
aacgggtaca gctcaccctc gtccagctac cgaaagaatg tagccaaaat ggtcaccgac 240
cgccatgcag ccgagtacaa catgcgccac aagaaccgcg gaatggcact gatcttcaac 300
catgagcact tcgaggtgcc caccttgaag tcccgcgcgg gaaccaatgt ggactgcgag 360
aatctgactc gggtgctcaa gcagctggac ttcgaagtga ccgtgtacaa ggactgccgc 420
tacaaggaca ttttgaggac aatcgagtat tcagcgtcgc agaatcacag cgatagtgat 480
tgcatcctgg tcgccatcct gtcgcacggc gagatgggct acatctacgc caaggacaca 540
cagtacaagc tggataacat ctggagcttc ttcacggcca atcactgccc ctcgctagcc 600
ggcaaaccca agttgttctt catacaggcc tgccagggcg acagattgga tggcggagtg 660
accatgcagc gttctcagac ggaaaccgat ggcgactcct cgatgagcta caagattcca 720
gtgcacgccg actttctgat cgcctactcg acggttcctg gattctattc gtggcgcaac 780
accacccgcg gcacgtggtt catgcagagc ctgtgcgccg aactggcggc caatggcaag 840
cggctggaca tcctgaccct gctcacattc gtgtgccagc gggtggccgt ggactttgag 900
tcctgcaccc cggacactcc ggagatgcac cagcagaagc agataccctg catcaccacc 960
atgctgacgc gcattctgcg tttcagcgac aagcagttgg ctccagccgg acgggtttga 1020
<210>361
<211>323
<212>PRT
<213>Drosophila melanogaster
<220>
<223>Caspase-1 precursor(insect)
C14.016
<400>361
Met Thr Asp Glu Cys Val Thr Arg Asn Tyr Gly Val Gly Ile Arg Ser
1 5 10 15
Pro Asn Gly Ser Glu Asn Arg Gly Ser Phe Ile Met Ala Asp Asn Thr
20 25 30
Asp Ala Lys Gly Cys Thr Pro Glu Ser Leu Val Val Gly Gly Ala Thr
35 40 45
Ala Ala Ser Pro Leu Pro Ala Asn Lys Phe Val Ala Arg Met Pro Val
50 55 60
Glu Arg Tyr Ala Ser Glu Tyr Asn Met Ser His Lys His Arg Gly Val
65 70 75 80
Ala Leu Ile Phe Asn His Glu Phe Phe Asp Ile Pro Ser Leu Lys Ser
85 90 95
Arg Thr Gly Thr Asn Val Asp Ala Gln Glu Leu Lys Lys Ala Phe Glu
100 105 110
Asn Leu Gly Phe Ala Val Ser Val His Lys Asp Cys Lys Leu Arg Asp
115 120 125
Ile Leu Lys His Val Gly Lys Ala Ala Glu Leu Asp His Thr Asp Asn
130 135 140
Asp Cys Leu Ala Val Ala Ile Leu Ser His Gly Glu His Gly Tyr Leu
145 150 155 160
Tyr Ala Lys Asp Thr Gln Tyr Lys Leu Asp Asn Ile Trp His Tyr Phe
165 170 175
Thr Ala Thr Phe Cys Pro Ser Leu Ala Gly Lys Pro Lys Leu Phe Phe
180 185 190
Ile Gln Ala Cys Gln Gly Asp Arg Leu Asp Gly Gly Ile Thr Leu Glu
195 200 205
Lys Gly Val Thr Glu Thr Asp Gly Glu Ser Ser Thr Ser Tyr Lys Ile
210 215 220
Pro Ile His Ala Asp Phe Leu Phe Ser Tyr Ser Thr Ile Pro Gly Tyr
225 230 235 240
Phe Ser Trp Arg Asn Ile Asn Asn Gly Ser Trp Tyr Mer Gln Ser Leu
245 250 255
Ile Arg Glu Leu Asn Ala Asn Gly Lys Lys Tyr Asp Leu Leu Thr Leu
260 265 270
Leu Thr Phe Val Asn Gln Arg Val Ala Leu Asp Phe Glu Ser Asn Val
275 280 285
Pro Ala Thr Pro Met Met Asp Arg Gln Lys Gln Ile Pro Cys Leu Thr
290 295 300
Ser Met Leu Thr Arg Ile Leu Arg Phe Gly Asp Lys Pro Asn Gly Asn
305 310 315 320
Lys Ala Gly
<210>362
<211>1757
<212>DNA
<213>Drosophila melanogaster
<220>
<223>Caspase-1 precursor(insect)C14.016
<400>362
agtttgtttt cgccgctcag agcacgctta acctccgtta attgattgat taaacacgtc 60
cgactgtgag ccataaaagg acattcgctc gatcgaaagc cgacaaaagc tggctgagaa 120
gcgcaaaaca gctgcgctgc gcaaagtttg gccaacgcag agcaatagaa aacagaagca 180
gtgtcagaat aaaacaagaa tcagaccaaa aagggcgcaa caactgcgga gagagaaaag 240
gaaggaaaga gcagcgcgtg acgagggagg agcaggaggg agcgaacaaa aggcaagagg 300
aaacacaagg agaaacgcaa acaatcaaaa caagaactta agcaagaaaa ggggcgcaag 360
atgaccgacg agtgcgtaac cagaaactac ggagtaggca ttcgcagccc aaatggatcg 420
gaaaatcggg gcagctttat aatggctgat aataccgatg ccaagggctg tacgccggag 480
tctcttgttg ttggtggtgc aactgcagcg agccccttgc cggccaacaa gtttgtggct 540
cgaatgcccg tggaacgata tgcaagcgaa tacaacatga gtcacaagca ccgcggagtt 600
gcgctgatct tcaaccacga attcttcgac ataccctcgc tgaagagccg caccggaaca 660
aatgtcgatg cccaagagct gaaaaaggcc tttgaaaacc tgggattcgc ggtgtccgtg 720
cacaaggact gcaagttgag ggacatcctg aagcacgtgg ggaaggccgc cgagttggac 780
cacacggaca acgactgcct tgcggtggcc atactctcgc acggggagca cggctacctg 840
tacgccaagg atacgcagta caagctggac aacatctggc actacttcac tgccaccttc 900
tgcccctcgc tggcgggcaa gccgaagctg ttcttcatcc aagcctgcca gggcgatcgc 960
ttggacggag gcatcactct ggagaagggc gttaccgaga cggacgggga gtcctcgacg 1020
agctacaaga tacccataca cgccgacttt ctcttctcct actcgaccat tccgggctac 1080
ttctcctggc gcaacatcaa caatggctcc tggtacatgc aatcgctgat ccgcgagctg 1140
aacgccaatg gcaaaaagta cgacctgctc accctgctta cattcgttaa ccagcgcgta 1200
gccctagact ttgagtcgaa cgtgcccgcc acaccgatga tggatcgcca gaagcaaata 1260
ccgtgcctca cctccatgct gacgcgcata ctgcgcttcg gcgacaagcc gaacggcaat 1320
aaggctggct aggaagagat ctcccttcga agaagaactt tcaacagagg ttgtacaacc 1380
atcgcccgca atcattcacg ttggattcca attcacttca agtttagttt tagctgacgc 1440
gtgcgattaa cccgatggat attgaagcac ccgactccct ttcattcatt cccctgcctt 1500
tcccatgccc atttcggcct tatcattgac gaaattcgtt acccatttac ccactagatg 1560
acttccgtat gcataagcga gaccttgcca atagaaaacc cacaaatttg agataagata 1620
atacaatgaa gatttataca tacatacata cttttatgta cacgcaaaca tttatatttg 1680
atatgaataa aaatgaatta aaaacataaa aaaaaaaaaa aaaaaaaaaa aaaaaacaaa 1740
aaaaaaaaaa aaaaaaa 1757
<210>363
<211>377
<212>PRT
<213>Bos taurus
<220>
<223>Caspase-13 precursor C14.017
<400>363
Met Ala Glu Asp Lys His Asn Lys Asn Pro Leu Lys Met Leu Glu Ser
1 5 10 15
Leu Gly Lys Glu Leu Ile Ser Gly Leu Leu Asp Asp Phe Val Glu Lys
20 25 30
Asn Val Leu Lys Leu Glu Glu Glu Glu Lys Lys Lys Ile Tyr Asp Ala
35 40 45
Lys Leu Gln Asp Lys Ala Arg Val Leu Val Asp Ser Ile Arg Gln Lys
50 55 60
Asn Gln Glu Ala Gly Gln Val Phe Val Gln Thr Phe Leu Asn Ile Asp
65 70 75 80
Lys Asn Ser Thr Ser Ile Lys Ala Pro Glu Glu Thr Val Ala Gly Pro
85 90 95
Asp Glu Ser Val Gly Ser Ala Ala Thr Leu Lys Leu Cys Pro His Glu
100 105 110
Glu Phe Leu Lys Leu Cys Lys Glu Arg Ala Gly Glu Ile Tyr Pro Ile
115 120 125
Lys Glu Arg Lys Asp Arg Thr Arg Leu Ala Leu Ile Ile Cys Asn Thr
130 135 140
Glu Phe Asp His Met Pro Pro Arg Asn Gly Ala Ala Leu Asp Ile Leu
145 150 155 160
Gly Met Lys Gln Leu Leu Glu Gly Leu Gly Tyr Thr Val Glu Val Glu
165 170 175
Glu Lys Leu Thr Ala Arg Asp Met Glu Ser Val Leu Trp Lys Phe Ala
180 185 190
Ala Arg Glu Glu His Lys Ser Ser Asp Ser Thr Phe Leu Val Phe Met
195 200 205
Ser His Gly Ile Leu Asp Gly Ile Cys Gly Thr Met His Ser Glu Glu
210 215 220
Glu Pro Asp Val Leu Pro Tyr Asp Thr Ile Phe Arg Thr Phe Asn Asn
225 230 235 240
Arg Asn Cys Leu Ser Leu Lys Asp Lys Pro Lys Val Ile Ile Val Gln
245 250 255
Ala Cys Arg Gly Ala Asn Arg Gly Glu Leu Trp Val Ser Asp Ser Pro
260 265 270
Pro Ala Leu Ala Asp Ser Phe Ser Gln Ser Ser Glu Asn Leu Glu Glu
275 280 285
Asp Ala Val Tyr Lys Thr His Val Glu Lys Asp Phe Ile Ala Phe Cys
290 295 300
Ser Ser Thr Pro His Asn Val Ser Trp Arg Asp Ile Lys Lys Gly Ser
305 310 315 320
Leu Phe Ile Thr Arg Leu Ile Thr Cys Phe Gln Lys Tyr Ala Trp Cys
325 330 335
Cys His Leu Glu Glu Val Phe Arg Lys Val Gln Gln Ser Phe Glu Lys
340 345 350
Pro Asn Val Lys Ala Gln Met Pro Thr Val Glu Arg Leu Ser Met Thr
355 360 365
Arg Tyr Phe Tyr Leu Phe Pro Gly Asn
370 375
<210>364
<211>2060
<212>DNA
<213>Homo sapiens
<220>
<223>Caspase-13 precursor C14.017
<400>364
ccacgcgtcc gcggacgcgt gggctccttc aaggcagtag gaagatacta agcttattat 60
ctatggctga agacaaacac aacaaaaacc cacttaagat gttggaatct ttgggcaaag 120
aactcatttc tggccttttg gatgactttg tggaaaaaaa tgtcctgaaa ttggaggaag 180
aggagaagaa aaaaatttat gatgccaaac ttcaagacaa agcccgggtc ttggttgatt 240
ctatacgaca gaaaaaccaa gaggcaggtc aagtctttgt tcaaactttc ctaaacatag 300
acaaaaattc caccagtata aaagctcctg aggaaactgt ggctggacca gatgagtcag 360
tgggatctgc agctaccctc aagctttgcc ctcatgaaga attcctgaaa ctgtgtaaag 420
aaagggctgg agagatctat ccaataaagg agagaaagga ccgcactcgt ctggctctca 480
tcatatgcaa cacagagttt gatcatatgc ctcccaggaa tggggctgcc cttgacatcc 540
ttggaatgaa gcagctgctt gagggtcttg gctacactgt ggaagtggaa gagaaactca 600
cagccaggga catggaatca gtgctgtgga aatttgctgc acgtgaagag cacaaatcct 660
cagacagtac attcttggtg ttcatgtctc atggcatcct ggatgggatc tgtgggacta 720
tgcacagtga ggaagaacca gatgtgttac cttatgatac catcttccgg acattcaaca 780
accgtaattg cctcagtcta aaggacaaac ctaaagtcat cattgtccag gcctgcagag 840
gtgcaaatcg tggggaattg tgggtcagtg actctccacc agccttggca gacagctttt 900
cacagtcatc cgagaacctg gaagaggatg ctgtttacaa gacccatgta gagaaggact 960
tcattgcttt ctgttcctca actccacata atgtgtcctg gagagacata aaaaaaggtt 1020
ctctcttcat tacacgactc atcacatgct tccaaaaata tgcttggtgc tgtcatcttg 1080
aggaagtatt taggaaggta caacaatcat ttgaaaaacc aaatgttaaa gcccagatgc 1140
ccactgttga acgactctct atgacaagat atttctacct ctttcctggc aactgaaaat 1200
aaaaatcaca ggaaattcaa ccatttatca gcttcaagaa gcatttttat cagcacagca 1260
tccatgttta accttttgtc tttcattaaa gtgaaaacat atgaactgtt ctttggggtc 1320
ctctaagaaa gaatagaatt tcaattaaaa caatggatgg atggaaataa agtagaagaa 1380
gaaaactgga ttttcttgtt atattgcata taatgcctgc actttactga gtgaagagaa 1440
ctagtcatga cttgccctca gcagcagggt gaaaggagat ggtgcacatc tggagacaag 1500
gaccaaaaac tgggtcacct gctcctttga tcactcatca aaccttgcaa ctagaattat 1560
ttggaagaat atttctaatt tattatttaa ccaacatcat aggtcaagtt caattttttt 1620
tcagaaagtt gttatggtac ctatgcatct ttcttcttat ccataaatta atttgctttc 1680
ttgttgatga tttttcattc cagatttcca agcttcagga aaattttgtt atttagcaaa 1740
cacttggtaa ttgtctaaaa taaattgggc ctttgattta agatggtata attgggtgtg 1800
cttgtataga aagacagaca ttattagata cagaacatcc aagagtgtgg gaaatgtcag 1860
caatgggaaa gaaaatgtta aattttctgg aaagtgaatg tcagtagagg aattttagca 1920
acatggccat ctgagccact gactaaggag atgtaccact tcaattacat acatgcagaa 1980
tgctgaatag atatatcagg acatgctgaa ctgtattaca aataaaatga ttctctagat 2040
atcaaaaaaa aaaaaaaaaa 2060
<210>365
<211>242
<212>PRT
<213>Homo sapiens
<220>
<223>Caspase-14 precursor C14.018
<400>365
Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly
1 5 10 15
Ala Arg Leu Ala Leu Ile Leu Cys Val Thr Lys Ala Arg Glu Gly Ser
20 25 30
Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gln Leu Arg Phe
35 40 45
Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gln Phe Gln Glu Glu
50 55 60
Leu Glu Lys Phe Gln Gln Ala Ile Asp Ser Arg Glu Asp Pro Val Ser
65 70 75 80
Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys
85 90 95
Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu
100 105 110
Asn Asn Lys Asn Cys Gln Ala Leu Arg Ala Lys Pro Lys Val Tyr Ile
115 120 125
Ile Gln Ala Cys Arg Gly Glu Gln Arg Asp Pro Gly Glu Thr Val Gly
130 135 140
Gly Asp Glu Ile Val Met Val Ile Lys Asp Ser Pro Gln Thr Ile Pro
145 150 155 160
Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr Ile
165 170 175
Ala Tyr Arg His Asp Gln Lys Gly Ser Cys Phe Ile Gln Thr Leu Val
180 185 190
Asp Val Phe Thr Lys Arg Lys Gly His Ile Leu Glu Leu Leu Thr Glu
195 200 205
Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gln Glu Gly Lys Ala
210 215 220
Arg Lys Thr Asn Pro Glu Ile Gln Ser Thr Leu Arg Lys Arg Leu Tyr
225 230 235 240
Leu Gln
<210>366
<211>777
<212>DNA
<213>Homo sapiens
<220>
<223>Caspase-14 precursor C14.018
<400>366
aggatcagac aagggtgctg agagccggga ctcacaacca aaggagaaat gagcaatccg 60
cggtctttgg aagaggagaa atatgatatg tcaggtgccc gcctggccct aatactgtgt 120
gtcaccaaag cccgggaagg ttccgaagaa gacctggatg ctctggaaca catgtttcgg 180
cagctgagat tcgaaagcac catgaaaaga gaccccactg ccgagcaatt ccaggaagag 240
ctggaaaaat tccagcaggc catcgattcc cgggaagatc ccgtcagttg tgccttcgtg 300
gtactcatgg ctcacgggag ggaaggcttc ctcaagggag aagatgggga gatggtcaag 360
ctggagaatc tcttcgaggc cctgaacaac aagaactgcc aggccctgcg agctaagccc 420
aaggtgtaca tcatacaggc ctgtcgagga gaacaaaggg accccggtga aacagtaggt 480
ggagatgaga ttgtgatggt catcaaagac agcccacaaa ccatcccaac atacacagat 540
gccttgcacg tttattccac ggtagaggga tacatcgcct accgacatga tcagaaaggc 600
tcatgcttta tccagaccct ggtggatgtg ttcacgaaga ggaaaggaca tatcttggaa 660
cttctgacag aggtgacccg gcggatggca gaagcagagc tggttcaaga aggaaaagca 720
aggaaaacga accctgaaat ccaaagcacc ctccggaaac ggctgtatct gcagtag 777
<210>367
<211>450
<212>PRT
<213>Drosophila melanogaster
<220>
<223>Caspase Nc precursor
C14.019
<400>367
Met Gln Pro Pro Glu Leu Glu Ile Gly Met Pro Lys Arg His Arg Glu
1 5 10 15
His Ile Arg Lys Asn Leu Asn Ile Leu Val Glu Trp Thr Asn Tyr Glu
20 25 30
Arg Leu Ala Met Glu Cys Val Gln Gln Gly Ile Leu Thr Val Gln Met
35 40 45
Leu Arg Asn Thr Gln Asp Leu Asn Gly Lys Pro Phe Asn Met Asp Glu
50 55 60
Lys Asp Val Arg Val Glu Gln His Arg Arg Leu Leu Leu Lys Ile Thr
65 70 75 80
Gln Arg Gly Pro Thr Ala Tyr Asn Leu Leu Ile Asn Ala Leu Arg Asn
85 90 95
Ile Asn Cys Leu Asp Ala Ala Val Leu Leu Glu Ser Val Asp Glu Ser
100 105 110
Asp Ser Arg Pro Pro Phe Ile Ser Leu Asn Glu Arg Arg Thr Ser Arg
115 120 125
Lys Ser Ala Asp Ile Val Asp Thr Pro Ser Pro Glu Ala Ser Glu Gly
130 135 140
Pro Cys Val Ser Lys Leu Arg Asn Glu Pro Leu Gly Ala Leu Thr Pro
145 150 155 160
Tyr Val Gly Val Val Asp Gly Pro Glu Val Lys Lys Ser Lys Lys Ile
165 170 175
His Gly Gly Asp Ser Ala Ile Leu Gly Thr Tyr Lys Met Gln Ser Arg
180 185 190
Phe Asn Arg Gly Val Leu Leu Met Val Asn Ile Met Asp Tyr Pro Asp
195 200 205
Gln Asn Arg Arg Arg Ile Gly Ala Glu Lys Asp Ser Lys Ser Leu Ile
210 215 220
His Leu Phe Gln Glu Leu Asn Phe Thr Ile Phe Pro Tyr Gly Asn Val
225 230 235 240
Asn Gln Asp Gln Phe Phe Lys Leu Leu Thr Met Val Thr Ser Ser Ser
245 250 255
Tyr Val Gln Asn Thr Glu Cys Phe Val Met Val Leu Met Thr His Gly
260 265 270
Asn Ser Val Glu Gly Lys Glu Lys Val Glu Phe Cys Asp Gly Ser Val
275 280 285
Val Asp Met Gln Lys Ile Lys Asp His Phe Gln Thr Ala Lys Cys Pro
290 295 300
Tyr Leu Val Asn Lys Pro Lys Val Leu Met Phe Pro Phe Cys Arg Gly
305 3l0 315 320
Asp Glu Tyr Asp Leu Gly His Pro Lys Asn Gln Gly Asn Leu Met Glu
325 330 335
Pro Val Tyr Thr Ala Gln Glu Glu Lys Trp Pro Asp Thr Gln Thr Glu
340 345 350
Gly Ile Pro Ser Pro Ser Thr Asn Val Pro Ser Leu Ala Asp Thr Leu
355 360 365
Val Cys Tyr Ala Asn Thr Pro Gly Tyr Val Thr His Arg Asp Leu Asp
370 375 380
Thr Gly Ser Trp Tyr Ile Gln Lys Phe Cys Gln Val Met Ala Asp His
385 390 395 400
Ala His Asp Thr Asp Leu Glu Asp Ile Leu Lys Lys Thr Ser Glu Ala
405 410 415
Val Gly Asn Lys Arg Thr Lys Lys Gly Ser Met Gln Thr Gly Ala Tyr
420 425 430
Asp Asn Leu Gly Phe Asn Lys Lys Leu Tyr Phe Asn Pro Gly Phe Phe
435 440 445
Asn Glu
450
<210>368
<211>2090
<212>DNA
<213>Drosophila melanogaster
<220>
<223>Caspase Nc precursor C14.019
<400>368
tttggcggcg cgaaacgctc gttggtcctt ttgtttcgaa agaaatccta ttcgaagatc 60
cccgatcctt gctaggatca tctagtgcaa tatatagact agttaattta cttttggaaa 120
aataaggaca ccagcagggc cgccgatttg tgcccctttc ttgaaagtcg caaaacaaaa 180
acaacgacga caacaacaaa gcggagacaa agaatcgaca agtagcgata aacgaaatca 240
ttcccgggaa aaccttggag acgggtgatt cactgccaat accactgcca attggagact 300
gatcacggca gccatccttg gcgcgcccaa taagcggagt caccggaacg cgtggaagcc 360
atatccggaa tgcagccgcc ggagctcgag attggaatgc cgaagaggca tcgcgagcat 420
atacgcaaga atctgaatat actggttgaa tggacgaact acgagcgtct ggccatggag 480
tgcgtgcaac agggcatcct aaccgttcag atgctaagaa atacgcagga tctcaatggc 540
aagccattca acatggacga gaaggatgtg cgtgtggagc agcatcgtag gctcctattg 600
aagatcaccc agcgtggtcc caccgcctat aacctgctga tcaatgcact gcgcaatatc 660
aattgtctgg atgcggccgt tctattggaa tccgtcgatg agtccgattc aaggccaccc 720
tttatctcgc taaacgaacg gagaaccagc cggaagtcgg ccgatattgt ggacacaccc 780
tcacccgaag cctccgaagg accctgcgtt agcaagctcc ggaatgagcc gctgggagca 840
ctcacccctt atgtgggtgt cgttgacggt cccgaggtaa aaaaatcgaa aaagatacat 900
ggtggggata gtgccatatt gggcacatat aagatgcaat cacgtttcaa ccgaggcgtt 960
ttgctaatgg ttaacataat ggactatccg gatcaaaacc gtcgacggat cggagccgaa 1020
aaggacagca agtcgttgat acacttgttt caagaactga attttacgat tttcccctat 1080
gggaacgtga atcaggatca gttctttaaa cttctgacaa tggtgacctc ctcgtcgtat 1140
gtgcagaata ccgagtgttt cgtaatggta ctgatgacac acggcaacag tgtggaggga 1200
aaagagaagg tggagttttg cgatggatct gtggtcgata tgcagaagat caaggaccat 1260
ttccagacgg ccaaatgtcc ttatttggtg aacaagccga aggtgcttat gtttcccttt 1320
tgccgcggcg atgaatatga tttgggccat ccaaagaatc aaggcaatct catggagcca 1380
gtgtatacgg cgcaagagga gaagtggcct gacacccaga cggagggcat acccagcccg 1440
agcaccaatg tgccaagtct tgccgacact ctggtctgct atgctaatac gccgggctat 1500
gttacccacc gcgatctcga cacgggcagc tggtacatcc agaagttttg ccaagtgatg 1560
gccgatcatg cccacgacac agaccttgag gatatcctaa agaagacgag cgaagccgtg 1620
ggtaataagc gcaccaagaa gggttccatg cagacaggtg cctatgataa tcttggcttt 1680
aataagaaac tctacttcaa tcccgggttt ttcaacgaat agttgccgcc actggacatt 1740
ttatcattcc ggatgcattt ttaaccgcat ttatgttctt atcgtcgcat ttagtatgtc 1800
ttttagatta tgtgttctgt gctcgcgtgc tataaaatgt ttatattgta acaaatactc 1860
attcaagtat tcttctaata cgtattgcat acctcattta aaggtacgaa attatgtaat 1920
ttatgtgata actgccagaa atctacaaat attataatac agagttccag tatatacaca 1980
gccagatttt tgaaatcgaa catttaggca aaatgtaaag caattttcca gtctgatttt 2040
tgagtgtgta cacttacaaa ataaacgatg atatttcaca aggcccaaag 2090
<210>369
<211>824
<212>PRT
<213>Homo sapiens
<220>
<223>MALT lymphoma translocation protein 1
(paracaspase)
C14.026
<400>369
Met Ser Leu Leu Gly Asp Pro Leu Gln Ala Leu Pro Pro Ser Ala Ala
1 5 10 15
Pro Thr Gly Pro Leu Leu Ala Pro Pro Ala Gly Ala Thr Leu Asn Arg
20 25 30
Leu Arg Glu Pro Leu Leu Arg Arg Leu Ser Glu Leu Leu Asp Gln Ala
35 40 45
Pro Glu Gly Arg Gly Trp Arg Arg Leu Ala Glu Leu Ala Gly Ser Arg
50 55 60
Gly Arg Leu Arg Leu Ser Cys Leu Asp Leu Glu Gln Cys Ser Leu Lys
65 70 75 80
Val Leu Glu Pro Glu Gly Ser Pro Ser Leu Cys Leu Leu Lys Leu Met
85 90 95
Gly Glu Lys Gly Cys Thr Val Thr Glu Leu Ser Asp Phe Leu Gln Ala
100 105 110
Met Glu His Thr Glu Val Leu Gln Leu Leu Ser Pro Pro Gly Ile Lys
115 120 125
Ile Thr Val Asn Pro Glu Ser Lys Ala Val Leu Ala Gly Gln Phe Val
130 135 140
Lys Leu Cys Cys Arg Ala Thr Gly His Pro Phe Val Gln Tyr Gln Trp
145 150 155 160
Phe Lys Met Asn Lys Glu Ile Pro Asn Gly Asn Thr Ser Glu Leu Ile
165 170 175
Phe Asn Ala Val His Val Lys Asp Ala Gly Phe Tyr Val Cys Arg Val
180 185 190
Asn Asn Asn Phe Thr Phe Glu Phe Ser Gln Trp Ser Gln Leu Asp Val
195 200 205
Cys Asp Ile Pro Glu Ser Phe Gln Arg Ser Val Asp Gly Val Ser Glu
210 215 220
Ser Lys Leu Gln Ile Cys Val Glu Pro Thr Ser Gln Lys Leu Met Pro
225 230 235 240
Gly Ser Thr Leu Val Leu Gln Cys Val Ala Val Gly Ser Pro Ile Pro
245 250 255
His Tyr Gln Trp Phe Lys Asn Glu Leu Pro Leu Thr His Glu Thr Lys
260 265 270
Lys Leu Tyr Met Val Pro Tyr Val Asp Leu Glu His Gln Gly Thr Tyr
275 280 285
Trp Cys His Val Tyr Asn Asp Arg Asp Ser Gln Asp Ser Lys Lys Val
290 295 300
Glu Ile Ile Ile Gly Arg Thr Asp Glu Ala Val Glu Cys Thr Glu Asp
305 310 315 320
Glu Leu Asn Asn Leu Gly His Pro Asp Asn Lys Glu Gln Thr Thr Asp
325 330 335
Gln Pro Leu Ala Lys Asp Lys Val Ala Leu Leu Ile Gly Asn Met Asn
340 345 350
Tyr Arg Glu His Pro Lys Leu Lys Ala Pro Leu Val Asp Val Tyr Glu
355 360 365
Leu Thr Asn Leu Leu Arg Gln Leu Asp Phe Lys Val Val Ser Leu Leu
370 375 380
Asp Leu Thr Glu Tyr Glu Met Arg Asn Ala Val Asp Glu Phe Leu Leu
385 390 395 400
Leu Leu Asp Lys Gly Val Tyr Gly Leu Leu Tyr Tyr Ala Gly His Gly
405 410 415
Tyr Glu Asn Phe Gly Asn Ser Phe Met Val Pro Val Asp Ala Pro Asn
420 425 430
Pro Tyr Arg Ser Glu Asn Cys Leu Cys Val Gln Asn Ile Leu Lys Leu
435 440 445
Met Gln Glu Lys Glu Thr Gly Leu Asn Val Phe Leu Leu Asp Met Cys
450 455 460
Arg Lys Arg Asn Asp Tyr Asp Asp Thr Ile Pro Ile Leu Asp Ala Leu
465 470 475 480
Lys Val Thr Ala Asn Ile Val Phe Gly Tyr Ala Thr Cys Gln Gly Ala
485 490 495
Glu Ala Phe Glu Ile Gln His Ser Gly Leu Ala Asn Gly Ile Phe Met
500 505 510
Lys Phe Leu Lys Asp Arg Leu Leu Glu Asp Lys Lys Ile Thr Val Leu
515 520 525
Leu Asp Glu Val Ala Glu Asp Met Gly Lys Cys His Leu Thr Lys Gly
530 535 540
Lys Gln Ala Leu Glu Ile Arg Ser Ser Leu Ser Glu Lys Arg Ala Leu
545 550 555 560
Thr Asp Pro Ile Gln Gly Thr Glu Tyr Ser Ala Glu Ser Leu Val Arg
565 570 575
Asn Leu Gln Trp Ala Lys Ala His Glu Leu Pro Glu Ser Met Cys Leu
580 585 590
Lys Phe Asp Cys Gly Val Gln Ile Gln Leu Gly Phe Ala Ala Glu Phe
595 600 605
Ser Asn Val Met Ile Ile Tyr Thr Ser Ile Val Tyr Lys Pro Pro Glu
610 615 620
Ile Ile Met Cys Asp Ala Tyr Val Thr Asp Phe Pro Leu Asp Leu Asp
625 630 635 640
Ile Asp Pro Lys Asp Ala Asn Lys Gly Thr Pro Glu Glu Thr Gly Ser
645 650 655
Tyr Leu Val Ser Lys Asp Leu Pro Lys His Cys Leu Tyr Thr Arg Leu
660 665 670
Ser Ser Leu Gln Lys Leu Lys Glu His Leu Val Phe Thr Val Cys Leu
675 680 685
Ser Tyr Gln Tyr Ser Gly Leu Glu Asp Thr Val Glu Asp Lys Gln Glu
690 695 700
Val Asn Val Gly Lys Pro Leu Ile Ala Lys Leu Asp Met His Arg Gly
705 710 715 720
Leu Gly Arg Lys Thr Cys Phe Gln Thr Cys Leu Met Ser Asn Gly Pro
725 730 735
Tyr Gln Ser Ser Ala Ala Thr Ser Gly Gly Ala Gly His Tyr His Ser
740 745 750
Leu Gln Asp Pro Phe His Gly Val Tyr His Ser His Pro Gly Asn Pro
755 760 765
Ser Asn Val Thr Pro Ala Asp Ser Cys His Cys Ser Arg Thr Pro Asp
770 775 780
Ala Phe Ile Ser Ser Phe Ala His His Ala Ser Cys His Phe Ser Arg
785 790 795 800
Ser Asn Val Pro Val Glu Thr Thr Asp Glu Ile Pro Phe Ser Phe Ser
805 810 815
Asp Arg Leu Arg Ile Ser Glu Lys
820
<210>370
<211>2828
<212>DNA
<213>Homo sapiens
<220>
<223>MALT lymphoma translocation protein 1
(paracaspase)C14.026
<400>370
ggggcgggga gcggacttcc tcctctgagg gccgtgccgc gctgccagat ttgttcttcc 60
gcccctgcct ccgcggctcg gaggcgagcg gaaggtgccc cggggccgag gcccgtgacg 120
gggcgggcgg gagccccggc agtccggggt cgccggcgag ggccatgtcg ctgttggggg 180
acccgctaca ggccctgccg ccctcggccg cccccacggg gccgctgctc gcccctccgg 240
ccggcgcgac cctcaaccgc ctgcgggagc cgctgctgcg gaggctcagc gagctcctgg 300
atcaggcgcc cgagggccgg ggctggagga gactggcgga gctggcgggg agtcgcgggc 360
gcctccgcct cagttgccta gacctggagc agtgttctct taaggtactg gagcctgaag 420
gaagccccag cctgtgtctg ctgaagttaa tgggtgaaaa aggttgcaca gtcacagaat 480
tgagtgattt cctgcaggct atggaacaca ctgaagttct tcagcttctc agccccccag 540
gaataaagat tactgtaaac ccagagtcaa aggcagtctt ggctggacag tttgtgaaac 600
tgtgttgccg ggcaactgga catccttttg ttcaatatca gtggttcaaa atgaataaag 660
agattccaaa tggaaataca tcagagctta tttttaatgc agtgcatgta aaagatgcag 720
gcttttatgt ctgtcgagtt aataacaatt tcacctttga attcagccag tggtcacagc 780
tggatgtttg cgacatccca gagagcttcc agagaagtgt tgatggcgtc tctgaatcca 840
agttgcaaat ctgtgttgaa ccaacttccc aaaagctgat gccaggcagc acattggttt 900
tacagtgtgt tgctgttgga agccctattc ctcactacca gtggttcaaa aatgaattac 960
cattaacaca tgagaccaaa aagctataca tggtgcctta tgtggatttg gaacaccaag 1020
gaacctactg gtgtcatgta tataatgatc gagacagtca agatagcaag aaggtagaaa 1080
tcatcatagg aagaacagat gaggcagtgg agtgcactga agatgaatta aataatcttg 1140
gtcatcctga taataaagag caaacaactg accagccttt ggcgaaggac aaggttgccc 1200
ttttgatagg aaatatgaat taccgggagc accccaagct caaagctcct ttggtggatg 1260
tgtacgaatt gactaactta ctgagacagc tggacttcaa agtggtttca ctgttggatc 1320
ttactgaata tgagatgcgt aatgctgtgg atgagttttt actcctttta gacaagggag 1380
tatatgggtt attatattat gcaggacatg gttatgaaaa ttttgggaac agcttcatgg 1440
tccccgttga tgctccaaat ccatataggt ctgaaaattg tctgtgtgta caaaatatac 1500
tgaaattgat gcaagaaaaa gaaactggac ttaatgtgtt cttattggat atgtgtagga 1560
aaagaaatga ctacgatgat accattccaa tcttggatgc actaaaagtc accgccaata 1620
ttgtgtttgg atatgccacg tgtcaaggag cagaagcttt tgaaatccag cattctggat 1680
tggcaaatgg aatctttatg aaatttttaa aagacagatt attagaagat aagaaaatca 1740
ctgtgttact ggatgaagtt gcagaagata tgggtaagtg tcaccttacc aaaggcaaac 1800
aggctctaga gattcgaagt agtttatctg agaagagagc acttactgat ccaatacagg 1860
gaacagaata ttctgctgaa tctcttgtgc ggaatctaca gtgggccaag gctcatgaac 1920
ttccagaaag tatgtgtctt aagtttgact gtggtgttca gattcaatta ggatttgcag 1980
ctgagttttc caatgtcatg atcatctata caagtatagt ttacaaacca ccggagataa 2040
taatgtgtga tgcctacgtt actgattttc cacttgatct agatattgat ccaaaagatg 2100
caaataaagg cacacctgaa gaaactggca gctacttggt atcaaaggat cttcccaagc 2160
attgcctcta taccagactc agttcactgc aaaaattaaa ggaacatcta gtcttcacag 2220
tatgtttatc atatcagtac tcaggattgg aagatactgt agaggacaag caggaagtga 2280
atgttgggaa acctctcatt gctaaattag acatgcatcg aggtttggga aggaagactt 2340
gctttcaaac ttgtcttatg tctaatggtc cttaccagag ttctgcagcc acctcaggag 2400
gagcagggca ttatcactca ttgcaagacc cattccatgg tgtttaccat tcacatcctg 2460
gtaatccaag taatgttaca ccagcagata gctgtcattg cagccggact ccagatgcat 2520
ttatttcaag tttcgctcac catgcttcat gtcattttag tagaagtaat gtgccagtag 2580
agacaactga tgaaatacca tttagtttct ctgacaggct cagaatttct gaaaaatgac 2640
ctccttgttt ttgaaagtta gcataatttt agatgcctgt gaaatagtac tgcacttaca 2700
taaagtgaga cattgtgaaa aggcaaattt gtatatgtag agaaagaata gtagtaactg 2760
tttcatagca aacttcagga ctttgagatg ttgaaattac attatttaat tacagacttc 2820
ctctttct 2828
<210>371
<211>480
<212>PRT
<213>Homo sapiens
<220>
<223>CASP8and FADD-like apoptosis regulator precursor
(c-FLIP)
C14.971
<400>371
Met Ser Ala Glu Val Ile His Gln Val Glu Glu Ala Leu Asp Thr Asp
1 5 10 15
Glu Lys Glu Met Leu Leu Phe Leu Cys Arg Asp Val Ala Ile Asp Val
20 25 30
Val Pro Pro Asn Val Arg Asp Leu Leu Asp Ile Leu Arg Glu Arg Gly
35 40 45
Lys Leu Ser Val Gly Asp Leu Ala Glu Leu Leu Tyr Arg Val Arg Arg
50 55 60
Phe Asp Leu Leu Lys Arg Ile Leu Lys Met Asp Arg Lys Ala Val Glu
65 70 75 80
Thr His Leu Leu Arg Asn Pro His Leu Val Ser Asp Tyr Arg Val Leu
85 90 95
Met Ala Glu Ile Gly Glu Asp Leu Asp Lys Ser Asp Val Ser Ser Leu
100 105 110
Ile Phe Leu Met Lys Asp Tyr Met Gly Arg Gly Lys Ile Ser Lys Glu
115 120 125
Lys Ser Phe Leu Asp Leu Val Val Glu Leu Glu Lys Leu Asn Leu Val
130 135 140
Ala Pro Asp Gln Leu Asp Leu Leu Glu Lys Cys Leu Lys Asn Ile His
145 150 155 160
Arg Ile Asp Leu Lys Thr Lys Ile Gln Lys Tyr Lys Gln Ser Val Gln
165 170 175
Gly Ala Gly Thr Ser Tyr Arg Asn Val Leu Gln Ala Ala Ile Gln Lys
180 185 190
Ser Leu Lys Asp Pro Ser Asn Asn Phe Arg Leu His Asn Gly Arg Ser
195 200 205
Lys Glu Gln Arg Leu Lys Glu Gln Leu Gly Ala Gln Gln Glu Pro Val
210 215 220
Lys Lys Ser Ile Gln Glu Ser Glu Ala Phe Leu Pro Gln Ser Ile Pro
225 230 235 240
Glu Glu Arg Tyr Lys Met Lys Ser Lys Pro Leu Gly Ile Cys Leu Ile
245 250 255
Ile Asp Cys Ile Gly Asn Glu Thr Glu Leu Leu Arg Asp Thr Phe Thr
260 265 270
Ser Leu Gly Tyr Glu Val Gln Lys Phe Leu His Leu Ser Met His Gly
275 280 285
Ile Ser Gln Ile Leu Gly Gln Phe Ala Cys Met Pro Glu His Arg Asp
290 295 300
Tyr Asp Ser Phe Val Cys Val Leu Val Ser Arg Gly Gly Ser Gln Ser
305 310 315 320
Val Tyr Gly Val Asp Gln Thr His Ser Gly Leu Pro Leu His His Ile
325 330 335
Arg Arg Met Phe Met Gly Asp Ser Cys Pro Tyr Leu Ala Gly Lys Pro
340 345 350
Lys Met Phe Phe Ile Gln Asn Tyr Val Val Ser Glu Gly Gln Leu Glu
355 360 365
Asp Ser Ser Leu Leu Glu Val Asp Gly Pro Ala Met Lys Asn Val Glu
370 375 380
Phe Lys Ala Gln Lys Arg Gly Leu Cys Thr Val His Arg Glu Ala Asp
385 390 395 400
Phe Phe Trp Ser Leu Cys Thr Ala Asp Met Ser Leu Leu Glu Gln Ser
405 410 415
His Ser Ser Pro Ser Leu Tyr Leu Gln Cys Leu Ser Gln Lys Leu Arg
420 425 430
Gln Glu Arg Lys Arg Pro Leu Leu Asp Leu His Ile Glu Leu Asn Gly
435 440 445
Tyr Met Tyr Asp Trp Asn Ser Arg Val Ser Ala Lys Glu Lys Tyr Tyr
450 455 460
Val Trp Leu Gln His Thr Leu Arg Lys Lys Leu Ile Leu Ser Tyr Thr
465 470 475 480
<210>372
<211>1443
<212>DNA
<213>Homo sapiens
<220>
<223>CASP8and FADD-like apoptosis regulator precursor
(c-FLIP)C14.971
<400>372
atgtctgctg aagtcatcca tcaggttgaa gaagcacttg atacagatga gaaggagatg 60
ctgctctttt tgtgccggga tgttgctata gatgtggttc cacctaatgt cagggacctt 120
ctggatattt tacgggaaag aggtaagctg tctgtcgggg acttggctga actgctctac 180
agagtgaggc gatttgacct gctcaaacgt atcttgaaga tggacagaaa agctgtggag 240
acccacctgc tcaggaaccc tcaccttgtt tcggactata gagtgctgat ggcagagatt 300
ggtgaggatt tggataaatc tgatgtgtcc tcattaattt tcctcatgaa ggattacatg 360
ggccgaggca agataagcaa ggagaagagt ttcttggacc ttgtggttga gttggagaaa 420
ctaaatctgg ttgccccaga tcaactggat ttattagaaa aatgcctaaa gaacatccac 480
agaatagacc tgaagacaaa aatccagaag tacaagcagt ctgttcaagg agcagggaca 540
agttacagga atgttctcca agcagcaatc caaaagagtc tcaaggatcc ttcaaataac 600
ttcaggctcc ataatgggag aagtaaagaa caaagactta aggaacagct tggcgctcaa 660
caagaaccag tgaagaaatc cattcaggaa tcagaagctt ttttgcctca gagcatacct 720
gaagagagat acaagatgaa gagcaagccc ctaggaatct gcctgataat cgattgcatt 780
ggcaatgaga cagagcttct tcgagacacc ttcacttccc tgggctatga agtccagaaa 840
ttcttgcatc tcagtatgca tggtatatcc cagattcttg gccaatttgc ctgtatgccc 900
gagcaccgag actacgacag ctttgtgtgt gtcctggtga gccgaggagg ctcccagagt 960
gtgtatggtg tggatcagac tcactcaggg ctccccctgc atcacatcag gaggatgttc 1020
atgggagatt catgccctta tctagcaggg aagccaaaga tgttttttat tcagaactat 1080
gtggtgtcag agggccagct ggaggacagc agcctcttgg aggtggatgg gccagcgatg 1140
aagaatgtgg aattcaaggc tcagaagcga gggctgtgca cagttcaccg agaagctgac 1200
ttcttctgga gcctgtgtac tgcggacatg tccctgctgg agcagtctca cagctcacca 1260
tccctgtacc tgcagtgcct ctcccagaaa ctgagacaag aaagaaaacg cccactcctg 1320
gatcttcaca ttgaactcaa tggctacatg tatgattgga acagcagagt ttctgccaag 1380
gagaaatatt atgtctggct gcagcacact ctgagaaaga aacttatcct ctcctacaca 1440
taa 1443
<210>373
<211>4874
<212>DNA
<213>Artificial Sequence
<220>
<223>pCMV4
<400>373
cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc 60
tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag 120
ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctgatc tatacattga 180
atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct 240
attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc 300
caatatgacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg 360
ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc 420
cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca 480
tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg 540
cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg 600
acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt 660
ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca 720
ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg 780
tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc 840
ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag 900
ctcgtttagt gaaccgtcag aattgttttt atttttaatt ttctttcaaa tacttccatc 960
gaattcagat ctggtaccac gcgtatcgat aagcttgcat gcctgcaggt cgactctaga 1020
ggatccccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ttggaagttg 1080
ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact 1140
aggtgtcctc tataatatta tggggtggag gggggtggta tggagcaagg ggcccaagtt 1200
gggaagacaa cctgtagggc ctgcggggtc tattcgggaa ccaagctgga gtgcagtggc 1260
acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc 1320
tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg 1380
gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat 1440
ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc 1500
tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac agcataggct 1560
acctgccatg gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg 1620
cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt ctgtggaatg 1680
tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 1740
tgcatctcaa ttagtcagca accaggtgtg gaaaagtccc caggctcccc agcaggcaga 1800
agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 1860
atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 1920
tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga 1980
ggcttttttg gaggcctagg cttttgcaaa aagctcctcg aggaactgaa aaaccag~a 2040
gttaactggt aagtttagtc tttttgtctt ttatttcagg tcccggatcc ggtggtggtg 2100
caaatcaaag aactgctcct cagtggatgt tgcctttact tctaggcctg tacggaagtg 2160
ttacttctgc tctaaaagct gctgcaggtc gactctaggg ccgcgtaccg agctcgaatt 2220
ccctatagtg agtcgtatta aattcgtaat catggtcata gctgtttcct gtgtgaaatt 2280
gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 2340
gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 2400
cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 2460
tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 2520
tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 2580
ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaggc 2640
cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 2700
ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 2760
aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 2820
tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt 2880
gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 2940
cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 3000
ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 3060
cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct 3120
gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 3180
cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 3240
tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 3300
ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 3360
aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 3420
atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 3480
ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 3540
tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 3600
agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 3660
taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 3720
tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 3780
cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 3840
ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 3900
tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 3960
tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 4020
cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 4080
tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 4140
gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 4200
tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 4260
atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 4320
tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 4380
cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg 4440
tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 4500
cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 4560
gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 4620
ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 4680
gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 4740
tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 4800
aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa caaaatatta 4860
acgtttacaa tttc 4874
<210>374
<211>35
<212>DNA
<213>Artificial Sequence
<220>
<223>primer 496
<400>374
acgtggccca ggcggccttt cagtgtggcc aaaag 35
<210>375
<211>32
<212>DNA
<213>Artificial Sequence
<220>
<223>primer 497
<400>375
tcctggccgg cctggccgag caggccattc tc 32
<210>376
<211>4681
<212>DNA
<213>Artificial Sequence
<220>
<223>pComb3H phagemid vector
<400>376
gggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct 60
cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg 120
agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact 180
ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac 240
cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga 300
gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga 360
aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca 420
ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa 480
atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 540
tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc 600
aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc 660
acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt 720
acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt 780
ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg 840
ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact 900
caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg 960
ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga 1020
aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg 1080
aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa 1140
tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac 1200
aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc 1260
cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca 1320
ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga 1380
gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta 1440
agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc 1500
atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 1560
cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 1620
cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 1680
cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 1740
tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact 1800
tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 1860
ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 1920
aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 1980
cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 2040
ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 2100
agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 2160
ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 2220
acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 2280
cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 2340
gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa 2400
tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt 2460
ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt 2520
aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg 2580
gataacaatt gaattcagga ggaatttaaa atgaaaaaga cagctatcgc gattgcagtg 2640
gcactggctg gtttcgctac cgtggcccag gcggccgagc tcgtgttgac ccaatctcca 2700
gcttctttgg ctgtgtctct agggcagagg gccaccatct cctgcagagc cagcgaaagt 2760
gttgataatt atggcattag ttttatgaac tggttccaac agaaaccagg acagccaccc 2820
aaactcctca tctatgctgc atccaaccta ggatccgggg tccctgccag gtttagtggc 2880
agtgggtctg ggacagactt cagtctcaac atccatccta tggaggagga agatactgca 2940
atgtatttct gtcagcaaag taaggaggtt ccgctcacgt tcggtgctgg gaccaaggtg 3000
gagctgaaac gggctgatgc tgcaccaact gtatccatct tcccaccatc cagtgagcag 3060
ttaacatctg gaggtgcctc agtcgtgtgc ttcttgaaca acttctaccc caaagacatc 3120
aatgtcaagt ggaagattga tggcagtgaa cgacaaaatg gcgtcctgaa cagttggact 3180
gatcaggaca gcaaagacag cacctacagc atgagcagca ccctcacgtt gaccaaggac 3240
gagtatgaac gacataacag ctatacctgt gaggccactc acaagacatc aacttcaccc 3300
attgtcaaga gcttcaacat gaatcaccac taatctagat aattaattag gaggaattta 3360
aaatgaaata cctattgcct acggcagccg ctggattgtt attactcgct gcccaaccag 3420
ccatggccga ggtgcagctg ctcgagcagc ctgggtctgt gctggtaagg cctggagctt 3480
cagtgaagct gtcctgcaag gcttctggct acaccttcac cagctcctgg atacactggg 3540
cgaagcagag gcctggacaa ggccttgagt ggattggaga gattcatcct aatagtggta 3600
atactaacta caatgagaag ttcaagggca aggccacact gactgtagac acatcctcca 3660
gcacagccta cgtggatctc agcagcctga catctgagga ctctgcggtc tattactgtg 3720
caagatggag gtacggtagt ccctactact ttgactactg gggccaaggc accactctca 3780
ctgtctcctc agccaaaacg acacccccat ctgtctatcc actggcccct ggatctgctg 3840
cccaaactaa ctccatggtg accctgggat gcctggtcaa gggctatttc cctgagccag 3900
tgacagtgac ctggaactct ggatccctgt ccagcggtgt gcacaccttc ccagctgtcc 3960
tgcagtctga cctctacact ctgagcagct cagtgactgt cccctccagc acctggccca 4020
gcgagaccgt cacctgcaac gttgcccacc cggccagcag caccaaggtg gacaagaaaa 4080
ttgtgcccca tcatactagt ggccaggccg gccaggaggg tggtggctct gagggtggcg 4140
gttctgaggg tggcggctct gagggaggcg gttccggtgg tggctctggt tccggtgatt 4200
ttgattatga aaagatggca aacgctaata agggggctat gaccgaaaat gccgatgaaa 4260
acgcgctaca gtctgacgct aaaggcaaac ttgattctgt cgctactgat tacggtgctg 4320
ctatcgatgg tttcattggt gacgtttccg gccttgctaa tggtaatggt gctactggtg 4380
attttgctgg ctctaattcc caaatggctc aagtcggtga cggtgataat tcacctttaa 4440
tgaataattt ccgtcaatat ttaccttccc tccctcaatc ggttgaatgt cgcccttttg 4500
tctttagcgc tggtaaacca tatgaatttt ctattgattg tgacaaaata aacttattcc 4560
gtggtgtctt tgcgtttctt ttatatgttg ccacctttat gtatgtattt tctacgtttg 4620
ctaacatact gcgtaataag gagtcttaag ctagctaatt aatttaagcg gccgcagatc 4680
t 4681
<210>377
<211>4361
<212>DNA
<213>Artificial Sequence
<220>
<223>PBR322
<400>377
ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa 60
ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg 120
caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt 180
gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata 240
tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg 300
ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac 360
cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac 420
aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca 480
cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg 540
actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct 600
caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat 660
gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt 720
cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct 780
ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct 840
tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa 900
acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt 960
cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc 1020
cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca 1080
tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc 1140
gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat 1200
tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg 1260
ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga 1320
attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac 1380
atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg 1440
tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg 1500
ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc 1560
tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg 1620
taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca 1680
ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga 1740
ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa 1800
cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt 1860
ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc 1920
aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt 1980
ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac 2040
gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac 2100
ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc 2160
agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc 2220
cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg 2280
tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 2340
gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 2400
ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 2460
acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 2520
cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 2580
caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 2640
gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 2700
tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 2760
aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 2820
ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 2880
cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 2940
tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 3000
tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 3060
ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 3120
aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 3180
aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 3240
aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 3300
gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 3360
gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 3420
caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 3480
ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 3540
attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 3600
ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 3660
gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 3720
ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 3780
tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 3840
gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 3900
cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 3960
gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 4020
tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 4080
ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 4140
gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 4200
tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 4260
catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 4320
ataaaaatag gcgtatcacg aggccctttc gtcttcaaga a 4361
<210>378
<211>5
<212>PRT
<213>Artificial Sequence
<220>
<223>wildtype RCL sequence from P4-P1
<400>378
Val Ser Ala Arg Met
1 5
<210>379
<211>5
<212>PRT
<213>Artificial Sequence
<220>
<223>mutant RCL sequence from P4-P1’
<400>379
Arg Arg Ala Arg Met
1 5
<210>380
<211>44
<212>DNA
<213>Artificial Sequence
<220>
<223>RRARM mutagenic primer
<400>380
ccacagctgt cataaggagg gccagaatgg cccccgagga gatc 44
<210>381
<211>18
<212>DNA
<213>Artificial Sequence
<220>
<223>primer 535
<400>381
cagctatcgc gattgcag 18
<210>382
<211>17
<212>DNA
<213>Artificial Sequence
<220>
<223>primer 5542
<400>382
gtgcgcagcc atcccgg 17
<210>383
<211>19
<212>DNA
<213>Artificial Sequence
<220>
<221>variation
<222>8
<223>n＝a，g，c，or t
<220>
<221>variation
<222>9
<223>n＝a，g，c，or t
<220>
<221>variation
<222>10
<223>s＝g or c
<220>
<223>Primer TC30
<400>383
gccctggnns gcggccatc 19
<210>384
<211>20
<212>DNA
<213>Artificial Sequence
<220>
<221>variation
<222>8
<223>n＝a，g，c，or t
<220>
<221>variation
<222>9
<223>n＝a，g，c，or t
<220>
<221>variation
<222>10
<223>s＝g or c
<220>
<223>Primer TC155
<400>384
ggagcagnns aaaatgactg 20
<210>385
<211>18
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer 717
<400>385
tttcagtgtg gccaaaag 18
<210>386
<211>18
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer 718
<400>386
cagagtcttt tggccaca 18
<210>387
<211>35
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer 850
<400>387
ggggtaccgc caccatgaga gccctgctgg cgcgc 35
<210>388
<211>33
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer 851
<400>388
gctctagatc atcagagggc caggccattc tct 33
<210>389
<211>5
<212>PRT
<213>Artificial Sequence
<220>
<223>mutant P4-P1’RCL loop sequence
<400>389
Pro Phe Gly Arg Ser
1 5
<210>390
<211>44
<212>DNA
<213>Artificial Sequence
<220>
<223>PFGRS mutagenesis primer
<400>390
ccacagctgt catacccttc ggcagaagcg cccccgagga gatc 44
<210>391
<211>245
<212>PRT
<213>Artificial Sequence
<220>
<223>mature chymotrypsin
<400>391
Cys Gly Val Pro Ala Ile His Pro Val Leu Ser Gly Leu Ser Arg Ile
1 5 10 15
Val Asn Gly Glu Asp Ala Val Pro Gly Ser Trp Pro Trp Gln Val Ser
20 25 30
Leu Gln Asp Lys Thr Gly Phe His Phe Cys Gly Gly Ser Leu Ile Ser
35 40 45
Glu Asp Trp Val Val Thr Ala Ala His Cys Gly Val Arg Thr Ser Asp
50 55 60
Val Val Val Ala Gly Glu Phe Asp Gln Gly Ser Asp Glu Glu Asn Ile
65 70 75 80
Gln Val Leu Lys Ile Ala Lys Val Phe Lys Asn Pro Lys Phe Ser Ile
85 90 95
Leu Thr Val Asn Asn Asp Ile Thr Leu Leu Lys Leu Ala Thr Pro Ala
100 105 110
Arg Phe Ser Gln Thr Val Ser Ala Val Cys Leu Pro Ser Ala Asp Asp
115 120 125
Asp Phe Pro Ala Gly Thr Leu Cys Ala Thr Thr Gly Trp Gly Lys Thr
130 135 140
Lys Tyr Asn Ala Asn Lys Thr Pro Asp Lys Leu Gln Gln Ala Ala Leu
145 150 155 160
Pro Leu Leu Ser Asn Ala Glu Cys Lys Lys Ser Trp Gly Arg Arg Ile
165 170 175
Thr Asp Val Met Ile Cys Ala Gly Ala Ser Gly Val Ser Ser Cys Met
180 185 190
Gly Asp Ser Gly Gly Pro Leu Val Cys Gln Lys Asp Gly Ala Trp Thr
195 200 205
Leu Val Gly Ile Val Ser Trp Gly Ser Asp Thr Cys Ser Thr Ser Ser
210 215 220
Pro Gly Val Tyr Ala Arg Val Thr Lys Leu Ile Pro Trp Val Gln Lys
225 230 235 240
Ile Leu Ala Ala Asn
245
<210>392
<211>10
<212>DNA
<213>Artificial Sequence
<220>
<223>AATTCTATGG
<400>392
aattctatgg 10
<210>393
<211>10
<212>DNA
<213>Artificial Sequence
<220>
<223>tgcaccatag
<400>393
tgcaccatag 10
<210>394
<211>13
<212>DNA
<213>Artificial Sequence
<220>
<223>ATGGAACCCTGAA
<400>394
atggaaccct gaa 13
<210>395
<211>19
<212>DNA
<213>Artificial Sequence
<220>
<223>AGCTTCAGGGTTCCATCAC-
<400>395
agcttcaggg ttccatcac 19
<210>396
<211>380
<212>PRT
<213>Artificial Sequence
<220>
<223>PAI-I with MET as cloned into pPAIST7HS
<400>396
Met Val His His Pro Pro Ser Tyr Val Ala His Leu Ala Ser Asp Phe
1 5 10 15
Gly Val Arg Val Phe Gln Gln Val Ala Gln Ala Ser Lys Asp Arg ASn
20 25 30
Val Val Phe Ser Pro Tyr Gly Val Ala Ser Val Leu Ala Met Leu Gln
35 40 45
Leu Thr Thr Gly Gly Glu Thr Gln Gln Gln Ile Gln Ala Ala Met Gly
50 55 60
Phe Lys Ile Asp Asp Lys Gly Met Ala Pro Ala Leu Arg His Leu Tyr
65 70 75 80
Lys Glu Leu Met Gly Pro Trp Asn Lys Asp Glu Ile Ser Thr Thr Asp
85 90 95
Ala Ile Phe Val Gln Arg Asp Leu Lys Leu Val Gln Gly Phe Met Pro
100 105 110
His Phe Phe Arg Leu Phe Arg Ser Thr Val Lys Gln Val Asp Phe Ser
115 120 125
Glu Val Glu Arg Ala Arg Phe Ile Ile Asn Asp Trp Val Lys Thr His
130 135 140
Thr Lys Gly Met Ile Ser Asn Leu Leu Gly Lys Gly Ala Val Asp Gln
145 150 155 160
Leu Thr Arg Leu Val Leu Val Asn Ala Leu Tyr Phe Asn Gly Gln Trp
165 170 175
Lys Thr Pro Phe Pro Asp Ser Ser Thr His Arg Arg Leu Phe His Lys
180 185 190
Ser Asp Gly Ser Thr Val Ser Val Pro Met Met Ala Gln Thr Asn Lys
195 200 205
Phe Asn Tyr Thr Glu Phe Thr Thr Pro Asp Gly His Tyr Tyr Asp Ile
210 215 220
Leu Glu Leu Pro Tyr His Gly Asp Thr Leu Ser Met Phe Ile Ala Ala
225 230 235 240
Pro Tyr Glu Lys Glu Val Pro Leu Ser Ala Leu Thr Asn Ile Leu Ser
245 250 255
Ala Gln Leu Ile Ser His Trp Lys Gly Asn Met Thr Arg Leu Pro Arg
260 265 270
Leu Leu Val Leu Pro Lys Phe Ser Leu Glu Thr Glu Val Asp Leu Arg
275 280 285
Lys Pro Leu Glu Asn Leu Gly Met Thr Asp Met Phe Arg Gln Phe Gln
290 295 300
Ala Asp Phe Thr Ser Leu Ser Asp Gln Glu Pro Leu His Val Ala Gln
305 310 315 320
Ala Leu Gln Lys Val Lys Ile Glu Val Asn Glu Ser Gly Thr Val Ala
325 330 335
Ser Ser Ser Thr Ala Val Ile Val Ser Ala Arg Met Ala Pro Glu Glu
340 345 350
Ile Ile Met Asp Arg Pro Phe Leu Phe Val Val Arg His Asn Pro Thr
355 360 365
Gly Thr Val Leu Phe Met Gly Gln Val Met Glu Pro
370 375 380
<210>397
<211>21
<212>PRT
<213>Artificial Sequence
<220>
<223>Manduca sexta serpin lB RSL loop
<400>397
Glu Gly Ala Glu Ala Ala Ala Ala Asn Ala Phe Gly Ile Val Pro Lys
1 5 10 15
Ser Leu Ile Leu Tyr
20
<210>398
<211>21
<212>PRT
<213>Artificial Sequence
<220>
<223>Manduca sexta serpin 1K RSL loop
<400>398
Glu Gly Ala Glu Ala Ala Ala Ala Asn Ala Phe Lys Ile Thr Thr Tyr
1 5 10 15
Ser Phe His Phe Val
20
<210>399
<211>21
<212>PRT
<213>Artificial Sequence
<220>
<223>1-antichymotrypsin RSL loop
<400>399
Glu Gly Thr Glu Ala Ser Ala Ala Thr Ala Val Lys Ile Thr Leu Leu
1 5 10 15
Ser Ala Leu Val Glu
20
<210>400
<211>21
<212>PRT
<213>Artificial Sequence
<220>
<223>Antithrombin-III RSL loop
<400>400
Glu Gly Ser Glu Ala Ala Ala Ser Thr Ala Val Val Ile Ala Gly Arg
1 5 10 15
Ser Leu Asn Pro Asn
20
<210>401
<211>21
<212>PRT
<213>Artificial Sequence
<220>
<223>PAI-II RSL loop
<400>401
Glu Gly Thr Glu Ala Ala Ala Gly Thr Gly Gly Val Met Thr Gly Arg
1 5 10 15
Thr Gly His Gly Gly
20
<210>402
<211>21
<212>PRT
<213>Artificial Sequence
<220>
<223>1-antitrypsin RSL loop
<400>402
Lys Gly Thr Glu Ala Ala Gly Ala Met Phe Leu Glu Ala Ile Pro Met
1 5 10 15
Ser Ile Pro Pro Glu
20
<210>403
<211>21
<212>PRT
<213>Artificial Sequence
<220>
<223>PAI-I RSL loop
<400>403
Ser Gly Thr Val Ala Ser Ser Ser Thr Ala Val Ile Val Ser Ala Arg
1 5 10 15
Met Ala Pro Glu Glu
20
<210>404
<211>21
<212>PRT
<213>Artificial Sequence
<220>
<223>PAI-III RSL loop
<400>404
Ser Gly Thr Arg Ala Ala Ala Ala Thr Gly Thr Ile Phe Thr Phe Arg
1 5 10 15
Ser Ala Arg Leu Asn
20
<210>405
<211>21
<212>PRT
<213>Artificial Sequence
<220>
<223>Ovalbumin RSL loop
<400>405
Ala Gly Arg Glu Val Val Gly Ser Ala Glu Ala Gly Val Asp Ala Ala
1 5 10 15
Ser Val Ser Glu Glu
20
<210>406
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>TNF-alpha cleavage sequence
<400>406
Ala Glu Ala Lys
1
<210>407
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>TNF-R1 cleavage sequence(1)
<400>407
Glu Asn Val Lys
1
<210>408
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>TNF-R1 cleavage sequence(2)
<400>408
Gly Thr Glu Asp
1
<210>409
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>TNF-R2 cleavage sequence(1)
<400>409
Ser Pro Thr Arg
1
<210>410
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>TNF-R2 cleavage sequence(2)
<400>410
Val Ser Thr Arg
1
<210>411
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>TNF-R2 cleavage sequence(3)
<400>411
Ser Thr Ser Phe
1
<210>412
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>HER-2 cleavage sequence(1)
<400>412
Lys Phe Pro Asp
1
<210>413
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>HER-2 cleavage sequence(2)
<400>413
Ala Glu Gln Arg
1
<210>414
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>EGFR cleavage sequence(1)
<400>414
Lys Tyr Ala Asp
1
<210>415
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>EGFR cleavage sequence(2)
<400>415
Asn Gly Pro Lys
1
<210>416
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>VEGFR-1 cleavage sequence(1)
<400>416
Ser Ser Ala Tyr
1
<210>417
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>VEGFR-1 cleavage sequence(2)
<400>417
Gly Thr Ser Asp
1
<210>418
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>VEGFR-2 cleavage sequence(1)
<400>418
Ala Gln Glu Lys
1
<210>419
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>VEGFR-2 cleavage sequence(2)
<400>419
Arg Ile Asp Tyr
1
<210>420
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C3 cleavage sequence(1)
<400>420
Arg Glu Phe Lys
1
<210>421
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C3 cleavage sequence(2)
<400>421
Gly Leu Ala Arg
1
<210>422
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C3 cleavage sequence(3)
<400>422
Arg Leu Gly Arg
1
<210>423
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C3 cleavage sequence(4)
<400>423
Ala Glu Gly Lys
1
<210>424
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C3 cleavage sequence(5)
<400>424
Gln His Ala Arg
1
<210>425
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C3 cleavage sequence(6)
<400>425
Leu Pro Ser Arg
1
<210>426
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C3 cleavage sequence(7)
<400>426
Ser Leu Leu Arg
1
<210>427
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C3 cleavage sequence(8)
<400>427
Leu Gly Leu Ala
1
<210>428
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C3 cleavage sequence(9)
<400>428
Leu Ser Val Val
1
<210>429
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C4 cleavage sequence(1)
<400>429
His Arg Gly Arg
1
<210>430
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C2 cleavage sequence(1)
<400>430
Gly Ala Thr Arg
1
<210>431
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C2 cleavage sequence(2)
<400>431
Ser Leu Gly Arg
1
<210>432
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>C2 cleavage sequence(3)
<400>432
Val Phe Ala Lys
1
<210>433
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>urokinase plasminogen activator(uPA)preproprotein
<400>433
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>434
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR73 F173I；F30I(CHYM SYSTEM)
<400>434
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Ile Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>435
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR81 L224A，I240V；L73A，I89V(CHYM SYSTEM)
<400>435
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>436
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR1 L224P；L73P(CHYM SYSTEM)
<400>436
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Pro
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>437
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR3 R378C；R217C(CHYM SYSTEM)
<400>437
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Ash Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Cys Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>438
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR4 L312P；L155P(CHYM SYSTEM)
<400>438
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Pro Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>439
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR7 S226P，I240V，I298T；S75P，I89V，I138T(CHYM
SYSTEM)
<400>439
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Pro Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Thr Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>440
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR32 E294G；E137G(CHYM SYSTEM)
<400>440
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Gly Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>441
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR36 R223G，L312P；R72G，L155P(CHYM SYSTEM)
<400>441
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Gly Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Pro Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>442
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR37 G290D；G133D(CHYM SYSTEM)
<400>442
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Asp Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>443
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR66 V317A；V160A(CHYM SYSTEM)
<400>443
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Ala Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>444
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR24 V185D；V38D(CHYM SYSTEM)
<400>444
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Asp Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>445
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>AR85 F289L，V317A；F132L，V160A(CHYM SYSTEM)
<400>445
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Leu Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Ala Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>446
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF2 F173T，L224A，I240V；F30T，L73A，I189V(CHYM SYSTEM)
<400>446
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Thr Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>447
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF6 F173L，L224A，I240V；F30L，L73A，I189V(CHYM SYSTEM
<400>447
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Leu Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>448
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF11 F173V，L224A，I240V；F30V，L73A，I189V(CHYM SYSTEM)
<400>448
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>449
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF17 F173G，L224A，I240V；F30G，L73A，I189V(CHYM SYSTEM)
<400>449
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Gly Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>450
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF16 F173L，L224A，I240V；F30L，L73A，I189V(CHYM SYSTEM)
<400>450
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Leu Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>451
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF33 L224A，I240V，L312V；L155V，L73A，I189V(CHYM SYSTEM)
<400>451
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Val Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>452
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF35 F173M，L224A，I240V；F30M，L73A，I189V(CHYM SYSTEM)
<400>452
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Sar Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Met Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>453
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF36 L224A，L312M；L155M，L73A，I189V(CHYM SYSTEM)
<400>453
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Met Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>454
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF37 F173M，L224A，I240V；F30M，L73A，I189V(CHYM SYSTEM)
<400>454
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Sar Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Met Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>455
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF43 F173L，L224A，I240V；F30L，L73A，I189V(CHYM SYSTEM)
<400>455
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Gln Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Sar Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Leu Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>456
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF47 F173V，L224A，I240V；F30V，L73A，I189V(CHYM SYSTEM)
<400>456
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>457
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF48 F173L，L224A，I240V，L312M；F30L，L155M，L73A，I189V(CHYM SYSTEM)
<400>457
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Leu Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Met Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>458
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF103 F173L，L224A，I240V；F30L，L73A，I189V(CHYM SYSTEM)
<400>458
Ser Asn Gln Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Leu Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>459
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>ARF115 F173G，L224A，I240V，L312M；F30G，L155M，L73A，I189V(CHYM SYSTEM)
<400>459
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Sar Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Gly Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Ala
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Met Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Sar Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu ValCys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Lau Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>460
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/Ia F164V；F21V(CHYM SYSTEM)
<400>460
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Sar Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Val Thr Thr Ile Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>461
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/Ib I167L；I24L(CHYM SYSTEM)
<400>461
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Leu Glu Asn Gln Pro Trp Phe Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>462
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/Ic F173V；F30V(CHYM SYSTEM)
<400>462
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>463
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/Id F173L；F30L(CHYM SYSTEM)
<400>463
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Leu Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>464
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/IIa F173V，Y209H；F30V，Y61aH(CHYM SYSTEM)
<400>464
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
His Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 303 355
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>465
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/IIb F173V，K233E；F30V，K82E(CHYM SYSTEM)
<400>465
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Glu Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>466
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/IIc F173V，K313T；F30V，K156T(CHYM SYSTEM)
<400>466
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Thr Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>467
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/IIIa F173V，K233E，V316A；F30V，K82E，V159A
(CHYM SYSTEM)
<400>467
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Mer Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Glu Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Ala Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>468
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/IIIb F173V，K233E，T186A，V316A；F30V，K82E，
39A，V159A(CHYM SYSTEM)
<400>468
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Ala Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Glu Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Ala Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>469
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/IIIc F173V，K233E，T315A，V316A；F30V，K82E，
T158A，V159A(CHYM SYSTEM)
<400>469
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Glu Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Ala Ala Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu AlaLeu
405 410
<210>470
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/IIId F173V，Y209H，K243E；F173V，Y61aH，K92E
(CHYM SYSTEM)
<400>470
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
His Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Glu Met Lys Phe Glu Val Glu Asn Leu Ile
225 230 235 240
Leu His Glu Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Val Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Lys Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>471
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/IVa F173V，K233E，V316A，G231E，I240V，K348E；
F173V，K82E，V159A，E80G，I89V，K187E(CHYM
SYSTEM)
<400>471
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Gly Met Glu Phe Glu Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Ala Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Glu Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>472
<211>411
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA/IVb F173V，K233E，V316A，G231E，E235K，I240V，
K348E；F173V，K82E，V159A，E80G，E89K，I89V，K187E
(CHYM；SYSTEM)
<400>472
Ser Asn Glu Leu His Gln Val Pro Ser Asn Cys Asp Cys Leu Asn Gly
1 5 10 15
Gly Thr Cys Val Ser Asn Lys Tyr Phe Ser Asn Ile His Trp Cys Asn
20 25 30
Cys Pro Lys Lys Phe Gly Gly Gln His Cys Glu Ile Asp Lys Ser Lys
35 40 45
Thr Cys Tyr Glu Gly Asn Gly His Phe Tyr Arg Gly Lys Ala Ser Thr
50 55 60
Asp Thr Met Gly Arg Pro Cys Leu Pro Trp Asn Ser Ala Thr Val Leu
65 70 75 80
Gln Gln Thr Tyr His Ala His Arg Ser Asp Ala Leu Gln Leu Gly Leu
85 90 95
Gly Lys His Asn Tyr Cys Arg Asn Pro Asp Asn Arg Arg Arg Pro Trp
100 105 110
Cys Tyr Val Gln Val Gly Leu Lys Pro Leu Val Gln Glu Cys Met Val
115 120 125
His Asp Cys Ala Asp Gly Lys Lys Pro Ser Ser Pro Pro Glu Glu Leu
130 135 140
Lys Phe Gln Cys Gly Gln Lys Thr Leu Arg Pro Arg Phe Lys Ile Ile
145 150 155 160
Gly Gly Glu Phe Thr Thr Ile Glu Asn Gln Pro Trp Val Ala Ala Ile
165 170 175
Tyr Arg Arg His Arg Gly Gly Ser Val Thr Tyr Val Cys Gly Gly Ser
180 185 190
Leu Ile Ser Pro Cys Trp Val Ile Ser Ala Thr His Cys Phe Ile Asp
195 200 205
Tyr Pro Lys Lys Glu Asp Tyr Ile Val Tyr Leu Gly Arg Ser Arg Leu
210 215 220
Asn Ser Asn Thr Gln Gly Gly Met Glu Phe Lys Val Glu Asn Leu Val
225 230 235 240
Leu His Lys Asp Tyr Ser Ala Asp Thr Leu Ala His His Asn Asp Ile
245 250 255
Ala Leu Leu Lys Ile Arg Ser Lys Glu Gly Arg Cys Ala Gln Pro Ser
260 265 270
Arg Thr Ile Gln Thr Ile Cys Leu Pro Ser Met Tyr Asn Asp Pro Gln
275 280 285
Phe Gly Thr Ser Cys Glu Ile Thr Gly Phe Gly Lys Glu Asn Ser Thr
290 295 300
Asp Tyr Leu Tyr Pro Glu Gln Leu Lys Met Thr Ala Val Lys Leu Ile
305 310 315 320
Ser His Arg Glu Cys Gln Gln Pro His Tyr Tyr Gly Ser Glu Val Thr
325 330 335
Thr Lys Met Leu Cys Ala Ala Asp Pro Gln Trp Glu Thr Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Cys Ser Leu Gln Gly Arg Met
355 360 365
Thr Leu Thr Gly Ile Val Ser Trp Gly Arg Gly Cys Ala Leu Lys Asp
370 375 380
Lys Pro Gly Val Tyr Thr Arg Val Ser His Phe Leu Pro Trp Ile Arg
385 390 395 400
Ser His Thr Lys Glu Glu Asn Gly Leu Ala Leu
405 410
<210>473
<211>299
<212>PRT
<213>Autographa californica nucleopolyhedrovirus
<220>
<223>P35peptide
<400>473
Met Cys Val Ile Phe Pro Val Glu Ile Asp Val Ser Gln Thr Ile Ile
1 5 10 15
Arg Asp Cys Gln Val Asp Lys Gln Thr Arg Glu Leu Val Tyr Ile Asn
20 25 30
Lys Ile Met Asn Thr Gln Leu Thr Lys Pro Val Leu Met Met Phe Asn
35 40 45
Ile Ser Gly Pro Ile Arg Ser Val Thr Arg Lys Asn Asn Asn Leu Arg
50 55 60
Asp Arg Ile Lys Ser Lys Val Asp Glu Gln Phe Asp Gln Leu Glu Arg
65 70 75 80
Asp Tyr Ser Asp Gln Met Asp Gly Phe His Asp Ser Ile Lys Tyr Phe
85 90 95
Lys Asp Glu His Tyr Ser Val Ser Cys Gln Asn Gly Ser Val Leu Lys
100 105 110
Ser Lys Phe Ala Lys Ile Leu Lys Ser His Asp Tyr Thr Asp Lys Lys
115 120 125
Ser Ile Glu Ala Tyr Glu Lys Tyr Cys Leu Pro Lys Leu Val Asp Glu
130 135 140
Arg Asn Asp Tyr Tyr Val Ala Val Cys Val Leu Lys Pro Gly Phe Glu
145 150 155 160
Asn Gly Ser Asn Gln Val Leu Ser Phe Glu Tyr Asn Pro Ile Gly Asn
165 170 175
Lys Val Ile Val Pro Phe Ala His Glu Ile Asn Asp Thr Gly Leu Tyr
180 185 190
Glu Tyr Asp Val Val Ala Tyr Val Asp Ser Val Gln Phe Asp Gly Glu
195 200 205
Gln Phe Glu Glu Phe Val Gln Ser Leu Ile Leu Pro Ser Ser Phe Lys
210 215 220
Asn Ser Glu Lys Val Leu Tyr Tyr Asn Glu Ala Ser Lys Asn Lys Ser
225 230 235 240
Met Ile Tyr Lys Ala Leu Glu Phe Thr Thr Glu Ser Ser Trp Gly Lys
245 250 255
Ser Glu Lys Tyr Asn Trp Lys Ile Phe Cys Asn Gly Phe Ile Tyr Asp
260 265 270
Lys Lys Ser Lys Val Leu Tyr Val Lys Leu His Asn Val Thr Ser Ala
275 280 285
Leu Asn Lys Asn Val Ile Leu Asn Thr Ile Lys
290 295
<210>474
<211>2105
<212>DNA
<213>Homo sapiens
<220>
<223>Full length u-PA
<400>474
atgagagccc tgctggcgcg cctgcttctc tgcgtcctgg tcgtgagcga ctccaaaggc 60
agcaatgaac ttcatcaagt tccatcgaac tgtgactgtc taaatggagg aacatgtgtg 120
tccaacaagt acttctccaa cattcactgg tgcaactgcc caaagaaatt cggagggcag 180
cactggtgaa atagataagt caaaaacctg ctatgagggg aatggtcact tttaccgagg 240
aaaggccagc actgacacca gttgccggcc ctgcctgccc tggaactctg ccactgtcct 300
tcagcaaacg taccatgccc acagatctga tgctcttcag ctgggcctgg ggaaacataa 360
ttactgcagg aacccagaca accggaggcg accctggtgc tatgtgcagg tgggcctaaa 420
gccgcttgtc caagagtgca tggtgcatga ctgcgcagat ggaaaaaagc cctcctctcc 480
tccagaagaa ttaaaatttc agtgtggcca aaagactctg aggccccgct ttgagattat 540
tgggggagaa ttcaccacca tcgagaacca gccctggttt gcggccatct acaggaggca 600
ccgggggggc tctgtcacct acgtgtgtgg aggcagcctc atcagccctt gctgggtgat 660
cagcgccaca cactgcttca ttgattaccc aaagaaggag gactacatcg tctacctggg 720
tcgctcaagg cttaactcca acacgcaagg ggagatgaag tttgaggtgg aaaacctcat 780
cctacacaag gactacagcg ctgacacgct tgctcaccac aacgacattg ccttgctgaa 840
gatccgttcc aaggagggca ggtgtgcgca gccatcccgg actatacaga ccatctgcct 900
gccctcgatg tataacgatc cccagtttgg cacaagctgt gagatcactg gctttggaaa 960
agagaattct accgactatc tctatccgga gcagctgaaa atgactgttg tgaagctgat 1020
ttcccaccgg gagtgtcagc agccccacta ctacggctct gaagtcacca ccaaaatgct 1080
atgtgctgct gacccccaat ggaaaacaga ttcctgccag ggagactcag ggggacccct 1140
cgtctgttcc ctccaaggcc gcatgacttt gactggaatt gtgagctggg gccgtggatg 1200
tgccctgaag gacaagccag gcgtctacac gagagtctca cacttcttac cctggatccg 1260
cagtcacacc aaggaagaga atggcctggc cctctgaggg ttttcagtgt ggccaaaaga 1320
ctctgaggcc ccgctttaag attattgggg gagaattcac caccatcgag aaccagccct 1380
ggtttgcggc catctacagg aggcaccggg ggggctctgt cacctacgtg tgtggaggca 1440
gcctcatcag cccttgctgg gtgatcagcg ccacacactg cttcattgat tacccaaaga 1500
aggaggacta catcgtctac ctgggtcgct caaggcttaa ctccaacacg caaggggaga 1560
tgaagtttga ggtggaaaac ctcatcctac acaaggacta cagcgctgac acgcttgctc 1620
accacaacga cattgccttg ctgaagatcc gttccaagga gggcaggtgt gcgcagccat 1680
cccggactat acagaccatc tgcctgccct cgatgtataa cgatccccag tttggcacaa 1740
gctgtgagat cactggcttt ggaaaagaga attctaccga ctatctctat ccggagcagc 1800
tgaaaatgac tgttgtgaag ctgatttccc accgggagtg tcagcagccc cactactacg 1860
gctctgaagt caccaccaaa atgctatgtg ctgctgaccc ccaatggaaa acagattcct 1920
gccagggaga ctcaggggga cccctcgtct gttccctcca aggccgcatg actttgactg 1980
gaattgtgag ctggggccgt ggatgtgccc tgaaggacaa gccaggcgtc tacacgagag 2040
tctcacactt cttaccctgg atccgcagtc acaccaagga agagaatggc ctgccctctg 2100
agggt 2105
<210>475
<211>804
<212>DNA
<213>Homo sapiens
<220>
<223>uPA protease domain
<400>475
tttcagtgtg gccaaaagac tctgaggccc cgctttaaga ttattggggg agaattcacc 60
accatcgaga accagccctg gtttgcggcc atctacagga ggcaccgggg gggctctgtc 120
acctacgtgt gtggaggcag cctcatcagc ccttgctggg tgatcagcgc cacacactgc 180
ttcattgatt acccaaagaa ggaggactac atcgtctacc tgggtcgctc aaggcttaac 240
tccaacacgc aaggggagat gaagtttgag gtggaaaacc tcatcctaca caaggactac 300
agcgctgaca cgcttgctca ccacaacgac attgccttgc tgaagatccg ttccaaggag 360
ggcaggtgtg cgcagccatc ccggactata cagaccatct gcctgccctc gatgtataac 420
gatccccagt ttggcacaag ctgtgagatc actggctttg gaaaagagaa ttctaccgac 480
tatctctatc cggagcagct gaaaatgact gttgtgaagc tgatttccca ccgggagtgt 540
cagcagcccc actactacgg ctctgaagtc accaccaaaa tgctatgtgc tgctgacccc 600
caatggaaaa cagattcctg ccagggagac tcagggggac ccctcgtctg ttccctccaa 660
ggccgcatga ctttgactgg aattgtgagc tggggccgtg gatgtgccct gaaggacaag 720
ccaggcgtct acacgagagt ctcacacttc ttaccctgga tccgcagtca caccaaggaa 780
gagaatggcc tgccctctga gggt 804
<210>476
<211>22
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer UPAF1
<400>476
atgagagccc tgctggcgcg cc 22
<210>477
<211>19
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer UPAF2
<400>477
ggaaaagaga attctaccg 19
<210>478
<211>6
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>478
Ile Ala Gly Arg Ser Leu
1 5
<210>479
<211>6
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>479
Ser Leu Gly Arg Lys Ile
1 5
<210>480
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>480
Val Leu Lys Asp
1
<210>481
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>481
Leu Val Glu Asp
1
<210>482
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>482
Trp Phe Lys Asp
1
<210>483
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>483
Arg Ile Tyr Asp
1
<210>484
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>484
Lys Val Gly Arg
1
<210>485
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>485
Arg Val Arg Lys
1
<210>486
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>486
Arg Lys Thr Lys
1
<210>487
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>487
Lys Thr Lys Lys
1
<210>488
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>488
Thr Lys Lys Arg
1
<210>489
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site
<400>489
Arg Arg Val Arg
1
<210>490
<211>1474
<212>PRT
<213>Homo sapiens
<220>
<223>Alpha-2-macroglobulin precursor(Alpha-2-M)
<400>490
Met Gly Lys Asn Lys Leu Leu His Pro Ser Leu Val Leu Leu Leu Leu
1 5 10 15
Val Leu Leu Pro Thr Asp Ala Ser Val Ser Gly Lys Pro Gln Tyr Met
20 25 30
Val Leu Val Pro Ser Leu Leu His Thr Glu Thr Thr Glu Lys Gly Cys
35 40 45
Val Leu Leu Ser Tyr Leu Asn Glu Thr Val Thr Val Ser Ala Ser Leu
50 55 60
Glu Ser Val Arg Gly Asn Arg Ser Leu Phe Thr Asp Leu Glu Ala Glu
65 70 75 80
Asn Asp Val Leu His Cys Val Ala Phe Ala Val Pro Lys Ser Ser Ser
85 90 95
Asn Glu Glu Val Met Phe Leu Thr Val Gln Val Lys Gly Pro Thr Gln
100 105 110
Glu Phe Lys Lys Arg Thr Thr Val Met Val Lys Asn Glu Asp Ser Leu
115 120 125
Val Phe Val Gln Thr Asp Lys Ser Ile Tyr Lys Pro Gly Gln Thr Val
130 135 140
Lys Phe Arg Val Val Ser Met Asp Glu Asn Phe His Pro Leu Asn Glu
145 150 155 160
Leu Ile Pro Leu Val Tyr Ile Gln Asp Pro Lys Gly Asn Arg Ile Ala
165 170 175
Gln Trp Gln Ser Phe Gln Leu Glu Gly Gly Leu Lys Gln Phe Ser Phe
180 185 190
Pro Leu Ser Ser Glu Pro Phe Gln Gly Ser Tyr Lys Val Val Val Gln
195 200 205
Lys Lys Ser Gly Gly Arg Thr Glu His Pro Phe Thr Val Glu Glu Phe
210 215 220
Val Leu Pro Lys Phe Glu Val Gln Val Thr Val Pro Lys Ile Ile Thr
225 230 235 240
Ile Leu Glu Glu Glu Met Asn Val Ser Val Cys Gly Leu Tyr Thr Tyr
245 250 255
Gly Lys Pro Val Pro Gly His Val Thr Val Ser Ile Cys Arg Lys Tyr
260 265 270
Ser Asp Ala Ser Asp Cys His Gly Glu Asp Ser Gln Ala Phe Cys Glu
275 280 285
Lys Phe Ser Gly Gln Leu Asn Ser His Gly Cys Phe Tyr Gln Gln Val
290 295 300
Lys Thr Lys Val Phe Gln Leu Lys Arg Lys Glu Tyr Glu Met Lys Leu
305 310 315 320
His Thr Glu Ala Gln Ile Gln Glu Glu Gly Thr Val Val Glu Leu Thr
325 330 335
Gly Arg Gln Ser Ser Glu Ile Thr Arg Thr Ile Thr Lys Leu Ser Phe
340 345 350
Val Lys Val Asp Ser His Phe Arg Gln Gly Ile Pro Phe Phe Gly Gln
355 360 365
Val Arg Leu Val Asp Gly Lys Gly Val Pro Ile Pro Asn Lys Val Ile
370 375 380
Phe Ile Arg Gly Asn Glu Ala Asn Tyr Tyr Ser Asn Ala Thr Thr Asp
385 390 395 400
Glu His Gly Leu Val Gln Phe Ser Ile Asn Thr Thr Asn Val Met Gly
405 410 415
Thr Ser Leu Thr Val Arg Val Asn Tyr Lys Asp Arg Ser Pro Cys Tyr
420 425 430
Gly Tyr Gln Trp Val Ser Glu Glu His Glu Glu Ala His His Thr Ala
435 440 445
Tyr Leu Val Phe Ser Pro Ser Lys Ser Phe Val His Leu Glu Pro Met
450 455 460
Ser His Glu Leu Pro Cys Gly His Thr Gln Thr Val Gln Ala His Tyr
465 470 475 480
Ile Leu Asn Gly Gly Thr Leu Leu Gly Leu Lys Lys Leu Ser Phe Tyr
485 490 495
Tyr Leu Ile Met Ala Lys Gly Gly Ile Val Arg Thr Gly Thr His Gly
500 505 510
Leu Leu Val Lys Gln Glu Asp Met Lys Gly His Phe Ser Ile Ser Ile
515 520 525
Pro Val Lys Ser Asp Ile Ala Pro Val Ala Arg Leu Leu Ile Tyr Ala
530 535 540
Val Leu Pro Thr Gly Asp Val Ile Gly Asp Ser Ala Lys Tyr Asp Val
545 550 555 560
Glu Asn Cys Leu Ala Asn Lys Val Asp Leu Ser Phe Ser Pro Ser Gln
565 570 575
Ser Leu Pro Ala Ser His Ala His Leu Arg Val Thr Ala Ala Pro Gln
580 585 590
Ser Val Cys Ala Leu Arg Ala Val Asp Gln Ser Val Leu Leu Met Lys
595 600 605
Pro Asp Ala Glu Leu Ser Ala Ser Ser Val Tyr Asn Leu Leu Pro Glu
610 615 620
Lys Asp Leu Thr Gly Phe Pro Gly Pro Leu Asn Asp Gln Asp Asp Glu
625 630 635 640
Asp Cys Ile Asn Arg His Asn Val Tyr Ile Asn Gly Ile Thr Tyr Thr
645 650 655
Pro Val Ser Ser Thr Asn Glu Lys Asp Met Tyr Ser Phe Leu Glu Asp
660 665 670
Met Gly Leu Lys Ala Phe Thr Asn Ser Lys Ile Arg Lys Pro Lys Met
675 680 685
Cys Pro Gln Leu Gln Gln Tyr Glu Met His Gly Pro Glu Gly Leu Arg
690 695 700
Val Gly Phe Tyr Glu Ser Asp Val Met Gly Arg Gly His Ala Arg Leu
705 710 715 720
Val His Val Glu Glu Pro His Thr Glu Thr Val Arg Lys Tyr Phe Pro
725 730 735
Glu Thr Trp Ile Trp Asp Leu Val Val Val Asn Ser Ala Gly Val Ala
740 745 750
Glu Val Gly Val Thr Val Pro Asp Thr Ile Thr Glu Trp Lys Ala Gly
755 760 765
Ala Phe Cys Leu Ser Glu Asp Ala Gly Leu Gly Ile Ser Ser Thr Ala
770 775 780
Ser Leu Arg Ala Phe Gln Pro Phe Phe Val Glu Leu Thr Met Pro Tyr
785 790 795 800
Ser Val Ile Arg Gly Glu Ala Phe Thr Leu Lys Ala Thr Val Leu Asn
805 810 815
Tyr Leu Pro Lys Cys Ile Arg Val Ser Val Gln Leu Glu Ala Ser Pro
820 825 830
Ala Phe Leu Ala Val Pro Val Glu Lys Glu Gln Ala Pro His Cys Ile
835 840 845
Cys Ala Asn Gly Arg Gln Thr Val Ser Trp Ala Val Thr Pro Lys Ser
850 855 860
Leu Gly Asn Val Asn Phe Thr Val Ser Ala Glu Ala Leu Glu Ser Gln
865 870 875 880
Glu Leu Cys Gly Thr Glu Val Pro Ser Val Pro Glu His Gly Arg Lys
885 890 895
Asp Thr Val Ile Lys Pro Leu Leu Val Glu Pro Glu Gly Leu Glu Lys
900 905 910
Glu Thr Thr Phe Asn Ser Leu Leu Cys Pro Ser Gly Gly Glu Val Ser
915 920 925
Glu Glu Leu Ser Leu Lys Leu Pro Pro Asn Val Val Glu Glu Ser Ala
930 935 940
Arg Ala Ser Val Ser Val Leu Gly Asp Ile Leu Gly Ser Ala Met Gln
945 950 955 960
Asn Thr Gln Asn Leu Leu Gln Met Pro Tyr Gly Cys Gly Glu Gln Asn
965 970 975
Met Val Leu Phe Ala Pro Asn Ile Tyr Val Leu Asp Tyr Leu Asn Glu
980 985 990
Thr Gln Gln Leu Thr Pro Glu Val Lys Ser Lys Ala Ile Gly Tyr Leu
995 1000 1005
Asn Thr Gly Tyr Gln Arg Gln Leu Asn Tyr Lys His Tyr Asp Gly Ser
1010 1015 1020
Tyr Ser Thr Phe Gly Glu Arg Tyr Gly Arg Asn Gln Gly Asn Thr Trp
1025 1030 1035 1040
Leu Thr Ala Phe Val Leu Lys Thr Phe Ala Gln Ala Arg Ala Tyr Ile
1045 1050 1055
Phe Ile Asp Glu Ala His Ile Thr Gln Ala Leu Ile Trp Leu Ser Gln
1060 1065 1070
Arg Gln Lys Asp Asn Gly Cys Phe Arg Ser Ser Gly Ser Leu Leu Asn
1075 1080 1085
Asn Ala Ile Lys Gly Gly Val Glu Asp Glu Val Thr Leu Ser Ala Tyr
1090 1095 1100
Ile Thr Ile Ala Leu Leu Glu Ile Pro Leu Thr Val Thr His Pro Val
1105 1110 1115 1120
Val Arg Asn Ala Leu Phe Cys Leu Glu Ser Ala Trp Lys Thr Ala Gln
1125 1130 1135
Glu Gly Asp His Gly Ser His Val Tyr Thr Lys Ala Leu Leu Ala Tyr
1140 1145 1150
Ala Phe Ala Leu Ala Gly Asn Gln Asp Lys Arg Lys Glu Val Leu Lys
1155 1160 1165
Ser Leu Asn Glu Glu Ala Val Lys Lys Asp Asn Ser Val His Trp Glu
1170 1175 1180
Arg Pro Gln Lys Pro Lys Ala Pro Val Gly His Phe Tyr Glu Pro Gln
1185 1190 1195 1200
Ala Pro Ser Ala Glu Val Glu Met Thr Ser Tyr Val Leu Leu Ala Tyr
1205 1210 1215
Leu Thr Ala Gln Pro Ala Pro Thr Ser Glu Asp Leu Thr Ser Ala Thr
1220 1225 1230
Asn Ile Val Lys Trp Ile Thr Lys Gln Gln Asn Ala Gln Gly Gly Phe
1235 1240 1245
Ser Ser Thr Gln Asp Thr Val Val Ala Leu His Ala Leu Ser Lys Tyr
1250 1255 1260
Gly Ala Ala Thr Phe Thr Arg Thr Gly Lys Ala Ala Gln Val Thr Ile
1265 1270 1275 1280
Gln Ser Ser Gly Thr Phe Ser Ser Lys Phe Gln Val Asp Asn Asn Asn
1285 1290 1295
Arg Leu Leu Leu Gln Gln Val Ser Leu Pro Glu Leu Pro Gly Glu Tyr
1300 1305 1310
Ser Met Lys Val Thr Gly Glu Gly Cys Val Tyr Leu Gln Thr Ser Leu
1315 1320 1325
Lys Tyr Asn Ile Leu Pro Glu Lys Glu Glu Phe Pro Phe Ala Leu Gly
1330 1335 1340
Val Gln Thr Leu Pro Gln Thr Cys Asp Glu Pro Lys Ala His Thr Ser
1345 1350 1355 1360
Phe Gln Ile Ser Leu Ser Val Ser Tyr Thr Gly Ser Arg Ser Ala Ser
1365 1370 1375
Asn Met Ala Ile Val Asp Val Lys Met Val Ser Gly Phe Ile Pro Leu
1380 1385 1390
Lys Pro Thr Val Lys Met Leu Glu Arg Ser Asn His Val Ser Arg Thr
1395 1400 1405
Glu Val Ser Ser Asn His Val Leu Ile Tyr Leu Asp Lys Val Ser Asn
1410 1415 1420
Gln Thr Leu Ser Leu Phe Phe Thr Val Leu Gln Asp Val Pro Val Arg
1425 1430 1435 1440
Asp Leu Lys Pro Ala Ile Val Lys Val Tyr Asp Tyr Tyr Glu Thr Asp
1445 1450 1455
Glu Phe Ala Ile Ala Glu Tyr Asn Ala Pro Cys Ser Lys Asp Leu Gly
1460 1465 1470
Asn Ala
<210>491
<211>439
<212>PRT
<213>Spodoptera litura nucleopolyhedrovirus
<400>491
Met Cys Val Leu Ile Pro Thr Phe Asp Ala Ser Ala Thr Thr Ile Val
1 5 10 15
Asp Cys Ala Asn Leu Ser Asp Ser Ser Leu Arg Asp Leu Ile Tyr Val
20 25 30
Asn Asn Val Ala Val Ser Lys Thr Gly Asn Tyr Ile Asn Arg Ser Val
35 40 45
Ile Met Ala Leu Asn Ile Ser Gly Pro Val Val Ser Val Asn Arg Val
50 55 60
Ser Met His Ile Val His Met Tyr Arg Ser His Ile Asp Arg Ile Phe
65 70 75 80
Asp Arg Phe Asn Arg Leu Thr Tyr Ser Ala Thr Val Thr Asp Gly Gly
85 90 95
Asp Ile Asp Arg Val Leu Lys Ser Asp Asp Tyr Thr Val Val Cys Met
100 105 110
Ser Arg Ser Glu Leu Leu Asp Asn Tyr Lys Lys Cys Leu Ile Asn Glu
115 120 125
Met Gly Ala Thr Gln Asp Asp Val Glu Lys Phe Arg Lys Tyr Cys Leu
130 135 140
Lys Pro Leu Val Glu Thr Glu Asn Asp Gly Gly Val Glu Asp Arg Lys
145 150 155 160
Lys Pro Tyr Val Val Ile Cys Ser Leu Lys Pro Lys Leu Leu Asn Lys
165 170 175
Asn Lys Thr Leu Cys Phe Thr Tyr Lys Pro Gln Thr Gly Gln Val Ile
180 185 190
Val Pro Leu Met His Glu Ile Asn Glu Ash Gly Ser Asp Val Tyr Ser
195 200 205
Tyr Glu Val Met Ala Met Ile Lys Asp Val Arg Leu Ser Ash Lys Pro
210 215 220
Leu Ser Met Leu Glu Arg Leu Lys Arg Ser Met Asp Gln Ile Val Leu
225 230 235 240
Asn His Asn Glu Asn Lys Tyr Met Met Thr Asn Gln Leu Glu Ser Leu
245 250 255
Lys Tyr Tyr Leu Lys Ser Ile Asn Ser Asn Glu Asp Asp Asp Leu Asn
260 265 270
Lys Leu Met Asn Val Leu Asn Asn Leu Ile Asp Gln Leu Glu Lys Val
275 280 285
Leu His Lys Lys Ser Ile Cys Asp Phe Ser Asp Asp Asp Glu Asp Asp
290 295 300
Asp Gly Leu Ile Asp Met Lys Tyr Tyr Asn Ser Ile Ile Glu Asp Asn
305 310 315 320
Met Arg Ile Leu Leu Met Lys Ile Gln Gln Tyr Ile Asp Asp Arg His
325 330 335
Met Phe Leu Ser Pro Pro Leu Ser Val Glu Lys Ser Thr Lys Ile Arg
340 345 350
Leu Phe Glu Asn Val Met Ser Phe Met Lys Arg Phe Asn Leu Gly Glu
355 360 365
Leu Tyr Ser Tyr Leu Lys Lys Ile Ile Asp Trp Lys Thr His Glu Asn
370 375 380
Asn Gln Lys Leu Ala Trp Pro Glu Ile Val Asp Asp Lys Tyr Ala Phe
385 390 395 400
Leu Lys Tyr Asp Tyr Phe Gly Thr Pro His Gly Phe Val Tyr Asp Gln
405 410 415
Lys Asp Arg Thr Met Tyr Val Lys Leu His Cys Gly Ile Ala Ala Asn
420 425 430
Arg Asn Ile Tyr Ile Asp Ile
435
<210>492
<211>341
<212>PRT
<213>Cowpox virus
<400>492
Met Asp Ile Phe Arg Glu Ile Ala Ser Ser Met Lys Gly Glu Asn Val
1 5 10 15
Phe Ile Ser Pro Pro Ser Ile Ser Ser Val Leu Thr Ile Leu Tyr Tyr
20 25 30
Gly Ala Asn Gly Ser Thr Ala Glu Gln Leu Ser Lys Tyr Val Glu Lys
35 40 45
Glu Ala Asp Lys Asn Lys Asp Asp Ile Ser Phe Lys Ser Met Asn Lys
50 55 60
Val Tyr Gly Arg Tyr Ser Ala Val Phe Lys Asp Ser Phe Leu Arg Lys
65 70 75 80
Ile Gly Asp Asn Phe Gln Thr Val Asp Phe Thr Asp Cys Arg Thr Val
85 90 95
Asp Ala Ile Asn Lys Cys Val Asp Ile Phe Thr Glu Gly Lys Ile Asn
100 105 110
Pro Leu Leu Asp Glu Pro Leu Ser Pro Asp Thr Cys Leu Leu Ala Ile
115 120 125
Ser Ala Val Tyr Phe Lys Ala Lys Trp Leu Met Pro Phe Glu Lys Glu
130 135 140
Phe Thr Ser Asp Tyr Pro Phe Tyr Val Ser Pro Thr Glu Met Val Asp
145 150 155 160
Val Ser Met Met Ser Met Tyr Gly Glu Ala Phe Asn His Ala Ser Val
165 170 175
Lys Glu Ser Phe Gly Asn Phe Ser Ile Ile Glu Leu Pro Tyr Val Gly
180 185 190
Asp Thr Ser Met Val Val Ile Leu Pro Asp Asn Ile Asp Gly Leu Glu
195 200 205
Ser Ile Glu Gln Asn Leu Thr Asp Thr Asn Phe Lys Lys Trp Cys Asp
210 215 220
Ser Met Asp Ala Met Phe Ile Asp Val His Ile Pro Lys Phe Lys Val
225 230 235 240
Thr Gly Ser Tyr Asn Leu Val Asp Ala Leu Val Lys Leu Gly Leu Thr
245 250 255
Glu Val Phe Gly Ser Thr Gly Asp Tyr Ser Asn Met Cys Asn Ser Asp
260 265 270
Val Ser Val Asp Ala Met Ile His Lys Thr Tyr Ile Asp Val Asn Glu
275 280 285
Glu Tyr Thr Glu Ala Ala Ala Ala Thr Cys Ala Leu Val Ala Asp Cys
290 295 300
Ala Ser Thr Val Thr Asn Glu Phe Cys Ala Asp His Pro Phe Ile Tyr
305 310 315 320
Val Ile Arg His Val Asp Gly Lys Ile Leu Phe Val Gly Arg Tyr Cys
325 330 335
Ser Pro Thr Thr Asn
340
<210>493
<211>432
<212>PRT
<213>Homo sapiens
<220>
<223>antithrombin III mature
<400>493
His Gly Ser Pro Val Asp Ile Cys Thr Ala Lys Pro Arg Asp Ile Pro
1 5 10 15
Met Asn Pro Met Cys Ile Tyr Arg Ser Pro Glu Lys Lys Ala Thr Glu
20 25 30
Asp Glu Gly Ser Glu Gln Lys Ile Pro Glu Ala Thr Asn Arg Arg Val
35 40 45
Trp Glu Leu Ser Lys Ala Asn Ser Arg Phe Ala Thr Thr Phe Tyr Gln
50 55 60
His Leu Ala Asp Ser Lys Asn Asp Asn Asp Asn Ile Phe Leu Ser Pro
65 70 75 80
Leu Ser Ile Ser Thr Ala Phe Ala Met Thr Lys Leu Gly Ala Cys Asn
85 90 95
Asp Thr Leu Gln Gln Leu Met Glu Val Phe Lys Phe Asp Thr Ile Ser
100 105 110
Glu Lys Thr Ser Asp Gln Ile His Phe Phe Phe Ala Lys Leu Asn Cys
115 120 125
Arg Leu Tyr Arg Lys Ala Asn Lys Ser Ser Lys Leu Val Ser Ala Asn
130 135 140
Arg Leu Phe Gly Asp Lys Ser Leu Thr Phe Asn Glu Thr Tyr Gln Asp
145 150 155 160
Ile Ser Glu Leu Val Tyr Gly Ala Lys Leu Gln Pro Leu Asp Phe Lys
165 170 175
Glu Asn Ala Glu Gln Ser Arg Ala Ala Ile Asn Lys Trp Val Ser Asn
180 185 190
Lys Thr Glu Gly Arg Ile Thr Asp Val Ile Pro Ser Glu Ala Ile Asn
195 200 205
Glu Leu Thr Val Leu Val Leu Val Asn Thr Ile Tyr Phe Lys Gly Leu
210 215 220
Trp Lys Ser Lys Phe Ser Pro Glu Asn Thr Arg Lys Glu Leu Phe Tyr
225 230 235 240
Lys Ala Asp Gly Glu Ser Cys Ser Ala Ser Met Met Tyr Gln Glu Gly
245 250 255
Lys Phe Arg Tyr Arg Arg Val Ala Glu Gly Thr Gln Val Leu Glu Leu
260 265 270
Pro Phe Lys Gly Asp Asp Ile Thr Met Val Leu Ile Leu Pro Lys Pro
275 280 285
Glu Lys Ser Leu Ala Lys Val Glu Lys Glu Leu Thr Pro Glu Val Leu
290 295 300
Gln Glu Trp Leu Asp Glu Leu Glu Glu Met Met Leu Val Val His Met
305 310 315 320
Pro Arg Phe Arg Ile Glu Asp Gly Phe Ser Leu Lys Glu Gln Leu Gln
325 330 335
Asp Met Gly Leu Val Asp Leu Phe Ser Pro Glu Lys Ser Lys Leu Pro
340 345 350
Gly Ile Val Ala Glu Gly Arg Asp Asp Leu Tyr Val Ser Asp Ala Phe
355 360 365
His Lys Ala Phe Leu Glu Val Asn Glu Glu Gly Ser Glu Ala Ala Ala
370 375 380
Ser Thr Ala Val Val Ile Ala Gly Arg Ser Leu Asn Pro Asn Arg Val
385 390 395 400
Thr Phe Lys Ala Asn Arg Pro Phe Leu Val Phe Ile Arg Glu Val Pro
405 410 415
Leu Asn Thr Ile Ile Phe Met Gly Arg Val Ala Asn Pro Cys Val Lys
420 425 430
<210>494
<211>9765
<212>DNA
<213>Artificial Sequence
<220>
<223>pAcGP67b vector
<400>494
aagctttact cgtaaagcga gttgaaggat catatttagt tgcgtttatg agataagatt 60
gaaagcacgt gtaaaatgtt tcccgcgcgt tggcacaact atttacaatg cggccaagtt 120
ataaaagatt ctaatctgat atgttttaaa acacctttgc ggcccgagtt gtttgcgtac 180
gtgactagcg aagaagatgt gtggaccgca gaacagatag taaaacaaaa ccctagtatt 240
ggagcaataa tcgatttaac caacacgtct aaatattatg atggtgtgca ttttttgcgg 300
gcgggcctgt tatacaaaaa aattcaagta cctggccaga ctttgccgcc tgaaagcata 360
gttcaagaat ttattgacac ggtaaaagaa tttacagaaa agtgtcccgg catgttggtg 420
ggcgtgcact gcacacacgg tattaatcgc accggttaca tggtgtgcag atatttaatg 480
cacaccctgg gtattgcgcc gcaggaagcc atagatagat tcgaaaaagc cagaggtcac 540
aaaattgaaa gacaaaatta cgttcaagat ttattaattt aattaatatt atttgcattc 600
tttaacaaat actttatcct attttcaaat tgttgcgctt cttccagcga accaaaacta 660
tgcttcgctt gctccgttta gcttgtagcc gatcagtggc gttgttccaa tcgacggtag 720
gattaggccg gatattctcc accacaatgt tggcaacgtt gatgttacgt ttatgctttt 780
ggttttccac gtacgtcttt tggccggtaa tagccgtaaa cgtagtgccg tcgcgcgtca 840
cgcacaacac cggatgtttg cgcttgtccg cggggtattg aaccgcgcga tccgacaaat 900
ccaccacttt ggcaactaaa tcggtgacct gcgcgtcttt tttctgcatt atttcgtctt 960
tcttttgcat ggtttcctgg aagccggtgt acatgcggtt tagatcagtc atgacgcgcg 1020
tgacctgcaa atctttggcc tcgatctgct tgtccttgat ggcaacgatg cgttcaataa 1080
actcttgttt tttaacaagt tcctcggttt tttgcgccac caccgcttgc agcgcgtttg 1140
tgtgctcggt gaatgtcgca atcagcttag tcaccaactg tttgctctcc tcctcccgtt 1200
gtttgatcgc gggatcgtac ttgccggtgc agagcacttg aggaattact tcttctaaaa 1260
gccattcttg taattctatg gcgtaaggca atttggactt cataatcagc tgaatcacgc 1320
cggatttagt aatgagcact gtatgcggct gcaaatacag cgggtcgccc cttttcacga 1380
cgctgttaga ggtagggccc ccattttgga tggtctgctc aaataacgat ttgtatttat 1440
tgtctacatg aacacgtata gctttatcac aaactgtata ttttaaactg ttagcgacgt 1500
ccttggccac gaaccggacc tgttggtcgc gctctagcac gtaccgcagg ttgaacgtat 1560
cttctccaaa tttaaattct ccaattttaa cgcgagccat tttgatacac gtgtgtcgat 1620
tttgcaacaa ctattgtttt ttaacgcaaa ctaaacttat tgtggtaagc aataattaaa 1680
tatgggggaa catgcgccgc tacaacactc gtcgttatga acgcagacgg cgccggtctc 1740
ggcgcaagcg gctaaaacgt gttgcgcgtt caacgcggca aacatcgcaa aagccaatag 1800
tacagttttg atttgcatat taacggcgat tttttaaatt atcttattta ataaatagtt 1860
atgacgccta caactccccg cccgcgttga ctcgctgcac ctcgagcagt tcgttgacgc 1920
cttcctccgt gtggccgaac acgtcgagcg ggtggtcgat gaccagcggc gtgccgcacg 1980
cgacgcacaa gtatctgtac accgaatgat cgtcgggcga aggcacgtcg gcctccaagt 2040
ggcaatattg gcaaattcga aaatatatac agttgggttg tttgcgcata tctatcgtgg 2100
cgttgggcat gtacgtccga acgttgattt gcatgcaagc cgaaattaaa tcattgcgat 2160
tagtgcgatt aaaacgttgt acatcctcgc ttttaatcat gccgtcgatt aaatcgcgca 2220
atcgagtcaa gtgatcaaag tgtggaataa tgttttcttt gtattcccga gtcaagcgca 2280
gcgcgtattt taacaaacta gccatcttgt aagttagttt catttaatgc aactttatcc 2340
aataatatat tatgtatcgc acgtcaagaa ttaacaatgc gcccgttgtc gcatctcaac 2400
acgactatga tagagatcaa ataaagcgcg aattaaatag cttgcgacgc aacgtgcacg 2460
atctgtgcac gcgttccggc acgagctttg attgtaataa gtttttacga agcgatgaca 2520
tgacccccgt agtgacaacg atcacgccca aaagaactgc cgactacaaa attaccgagt 2580
atgtcggtga cgttaaaact attaagccat ccaatcgacc gttagtcgaa tcaggaccgc 2640
tggtgcgaga agccgcgaag tatggcgaat gcatcgtata acgtgtggag tccgctcatt 2700
agagcgtcat gtttagacaa gaaagctaca tatttaattg atcccgatga ttttattgat 2760
aaattgaccc taactccata cacggtattc tacaatggcg gggttttggt caaaatttcc 2820
ggactgcgat tgtacatgct gttaacggct ccgcccacta ttaatgaaat taaaaattcc 2880
aattttaaaa aacgcagcaa gagaaacatt tgtatgaaag aatgcgtaga aggaaagaaa 2940
aatgtcgtcg acatgctgaa caacaagatt aatatgcctc cgtgtataaa aaaaatattg 3000
aacgatttga aagaaaacaa tgtaccgcgc ggcggtatgt acaggaagag gtttatacta 3060
aactgttaca ttgcaaacgt ggtttcgtgt gccaagtgtg aaaaccgatg tttaatcaag 3120
gctctgacgc atttctacaa ccacgactcc aagtgtgtgg gtgaagtcat gcatctttta 3180
atcaaatccc aagatgtgta taaaccacca aactgccaaa aaatgaaaac tgtcgacaag 3240
ctctgtccgt ttgctggcaa ctgcaagggt ctcaatccta tttgtaatta ttgaataata 3300
aaacaattat aaatgctaaa tttgtttttt attaacgata caaaccaaac gcaacaagaa 3360
catttgtagt attatctata attgaaaacg cgtagttata atcgctgagg taatatttaa 3420
aatcattttc aaatgattca cagttaattt gcgacaatat aattttattt tcacataaac 3480
tagacgcctt gtcgtcttct tcttcgtatt ccttctcttt ttcatttttc tcctcataaa 3540
aattaacata gttattatcg tatccatata tgtatctatc gtatagagta aattttttgt 3600
tgtcataaat atatatgtct tttttaatgg ggtgtatagt accgctgcgc atagtttttc 3660
tgtaatttac aacagtgcta ttttctggta gttcttcgga gtgtgttgct ttaattatta 3720
aatttatata atcaatgaat ttgggatcgt cggttttgta caatatgttg ccggcatagt 3780
acgcagcttc ttctagttca attacaccat tttttagcag caccggatta acataacttt 3840
ccaaaatgtt gtacgaaccg ttaaacaaaa acagttcacc tcccttttct atactattgt 3900
ctgcgagcag ttgtttgttg ttaaaaataa cagccattgt aatgagacgc acaaactaat 3960
atcacaaact ggaaatgtct atcaatatat agttgctgat atcatggaga taattaaaat 4020
gataaccatc tcgcaaataa ataagtattt tactgttttc gtaacagttt tgtaataaaa 4080
aaacctataa atattccgga ttattcatac cgtcccacca tcgggcgcgg atctatgcta 4140
ctagtaaatc agtcacacca aggcttcaat aaggaacaca caagcaagat ggtaagcgct 4200
attgttttat atgtgctttt ggcggcggcg gcgcattctg cctttgcggc ggatcttgga 4260
tcccgggcca tgggaattcc ggagcggccg ctgcagatct gatcctttcc tgggacccgg 4320
caagaaccaa aaactcactc tcttcaagga aatccgtaat gttaaacccg acacgatgaa 4380
gcttgtcgtt ggatggaaag gaaaagagtt ctacagggaa acttggaccc gcttcatgga 4440
agacagcttc cccattgtta acgaccaaga agtgatggat gttttccttg ttgtcaacat 4500
gcgtcccact agacccaacc gttgttacaa attcctggcc caacacgctc tgcgttgcga 4560
ccccgactat gtacctcatg acgtgattag gatcgtcgag ccttcatggg tgggcagcaa 4620
caacgagtac cgcatcagcc tggctaagaa gggcggcggc tgcccaataa tgaaccttca 4680
ctctgagtac accaactcgt tcgaacagtt catcgatcgt gtcatctggg agaacttcta 4740
caagcccatc gtttacatcg gtaccgactc tgctgaagag gaggaaattc tccttgaagt 4800
ttccctggtg ttcaaagtaa aggagtttgc accagacgca cctctgttca ctggtccggc 4860
gtattaaaac acgatacatt gttattagta catttattaa gcgctagatt ctgtgcgttg 4920
ttgatttaca gacaattgtt gtacgtattt taataattca ttaaatttat aatctttagg 4980
gtggtatgtt agagcgaaaa tcaaatgatt ttcagcgtct ttatatctga atttaaatat 5040
taaatcctca atagatttgt aaaataggtt tcgattagtt tcaaacaagg gttgtttttc 5100
cgaaccgatg gctggactat ctaatggatt ttcgctcaac gccacaaaac ttgccaaatc 5160
ttgtagcagc aatctagctt tgtcgatatt cgtttgtgtt ttgttttgta ataaaggttc 5220
gacgtcgttc aaaatattat gcgcttttgt atttctttca tcactgtcgt tagtgtacaa 5280
ttgactcgac gtaaacacgt taaataaagc ttggacatat ttaacatcgg gcgtgttagc 5340
tttattaggc cgattatcgt cgtcgtccca accctcgtcg ttagaagttg cttccgaaga 5400
cgattttgcc atagccacac gacgcctatt aattgtgtcg gctaacacgt ccgcgatcaa 5460
atttgtagtt gagctttttg gaattatttc tgattgcggg cgtttttggg cgggtttcaa 5520
tctaactgtg cccgatttta attcagacaa cacgttagaa agcgatggtg caggcggtgg 5580
taacatttca gacggcaaat ctactaatgg cggcggtggt ggagctgatg ataaatctac 5640
catcggtgga ggcgcaggcg gggctggcgg cggaggcgga ggcggaggtg gtggcggtga 5700
tgcagacggc ggtttaggct caaatgtctc tttaggcaac acagtcggca cctcaactat 5760
tgtactggtt tcgggcgccg tttttggttt gaccggtctg agacgagtgc gatttttttc 5820
gtttctaata gcttccaaca attgttgtct gtcgtctaaa ggtgcagcgg gttgaggttc 5880
cgtcggcatt ggtggagcgg gcggcaattc agacatcgat ggtggtggtg gtggtggagg 5940
cgctggaatg ttaggcacgg gagaaggtgg tggcggcggt gccgccggta taatttgttc 6000
tggtttagtt tgttcgcgca cgattgtggg caccggcgca ggcgccgctg gctgcacaac 6060
ggaaggtcgt ctgcttcgag gcagcgcttg gggtggtggc aattcaatat tataattgga 6120
atacaaatcg taaaaatctg ctataagcat tgtaatttcg ctatcgttta ccgtgccgat 6180
atttaacaac cgctcaatgt aagcaattgt attgtaaaga gattgtctca agctccgcac 6240
gccgataaca agccttttca tttttactac agcattgtag tggcgagaca cttcgctgtc 6300
gtcgacgtac atgtatgctt tgttgtcaaa aacgtcgttg gcaagcttta aaatatttaa 6360
aagaacatct ctgttcagca ccactgtgtt gtcgtaaatg ttgtttttga taatttgcgc 6420
ttccgcagta tcgacacgtt caaaaaattg atgcgcatca attttgttgt tcctattatt 6480
gaataaataa gattgtacag attcatatct acgattcgtc atggccacca caaatgctac 6540
gctgcaaacg ctggtacaat tttacgaaaa ctgcaaaaac gtcaaaactc ggtataaaat 6600
aatcaacggg cgctttggca aaatatctat tttatcgcac aagcccacta gcaaattgta 6660
tttgcagaaa acaatttcgg cgcacaattt taacgctgac gaaataaaag ttcaccagtt 6720
aatgagcgac cacccaaatt ttataaaaat ctattttaat cacggttcca tcaacaacca 6780
agtgatcgtg atggactaca ttgactgtcc cgatttattt gaaacactac aaattaaagg 6840
cgagctttcg taccaacttg ttagcaatat tattagacag ctgtgtgaag cgctcaacga 6900
tttgcacaag cacaatttca tacacaacga cataaaactc gaaaatgtct tatatttcga 6960
agcacttgat cgcgtgtatg tttgcgatta cggattgtgc aaacacgaaa actcacttag 7020
cgtgcacgac ggcacgttgg agtattttag tccggaaaaa attcgacaca caactatgca 7080
cgtttcgttt gactggtacg cggcgtgtta acatacaagt tgctaaccgg cggttcgtaa 7140
tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 7200
cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 7260
attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 7320
tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 7380
ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 7440
gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 7500
ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 7560
cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 7620
ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 7680
accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 7740
catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 7800
gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 7860
tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 7920
agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 7980
actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 8040
gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 8100
aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 8160
gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 8220
aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 8280
atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 8340
gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 8400
atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 8460
ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 8520
cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 8580
agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 8640
cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 8700
tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 8760
agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 8820
gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 8880
gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 8940
ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 9000
tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 9060
tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 9120
gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 9180
caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 9240
atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 9300
gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc 9360
tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag 9420
acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca 9480
gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg 9540
agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc 9600
aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 9660
tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg 9720
ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgcc 9765
<210>495
<211>8
<212>PRT
<213>Artificial Sequence
<220>
<223>FLAG tag
<400>495
Asp Tyr Lys Asp Asp Asp Asp Lys
1 5
<210>496
<211>6
<212>PRT
<213>Artificial Sequence
<220>
<223>6xHis tag
<400>496
His His His His His His
1 5
<210>497
<211>432
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant AT3 sequence
<223>AT3/RRVRKE
<400>497
His Gly Ser Pro Val Asp Ile Cys Thr Ala Lys Pro Arg Asp Ile Pro
1 5 10 15
Met Asn Pro Met Cys Ile Tyr Arg Ser Pro Glu Lys Lys Ala Thr Glu
20 25 30
Asp Glu Gly Ser Glu Gln Lys Ile Pro Glu Ala Thr Asn Arg Arg Val
35 40 45
Trp Glu Leu Ser Lys Ala Asn Ser Arg Phe Ala Thr Thr Phe Tyr Gln
50 55 60
His Leu Ala Asp Ser Lys Asn Asp Asn Asp Asn Ile Phe Leu Ser Pro
65 70 75 80
Leu Ser Ile Ser Thr Ala Phe Ala Met Thr Lys Leu Gly Ala Cys Asn
85 90 95
Asp Thr Leu Gln Gln Leu Met Glu Val Phe Lys Phe Asp Thr Ile Ser
100 105 110
Glu Lys Thr Ser Asp Gln Ile His Phe Phe Phe Ala Lys Leu Asn Cys
115 120 125
Arg Leu Tyr Arg Lys Ala Asn Lys Ser Ser Lys Leu Val Ser Ala Asn
130 135 140
Arg Leu Phe Gly Asp Lys Ser Leu Thr Phe Asn Glu Thr Tyr Gln Asp
145 150 155 160
Ile Ser Glu Leu Val Tyr Gly Ala Lys Leu Gln Pro Leu Asp Phe Lys
165 170 175
Glu Asn Ala Glu Gln Ser Arg Ala Ala Ile Asn Lys Trp Val Ser Asn
180 185 190
Lys Thr Glu Gly Arg Ile Thr Asp Val Ile Pro Ser Glu Ala Ile Asn
195 200 205
Glu Leu Thr Val Leu Val Leu Val Asn Thr Ile Tyr Phe Lys Gly Leu
210 215 220
Trp Lys Ser Lys Phe Ser Pro Glu Asn Thr Arg Lys Glu Leu Phe Tyr
225 230 235 240
Lys Ala Asp Gly Glu Ser Cys Ser Ala Ser Met Met Tyr Gln Glu Gly
245 250 255
Lys Phe Arg Tyr Arg Arg Val Ala Glu Gly Thr Gln Val Leu Glu Leu
260 265 270
Pro Phe Lys Gly Asp Asp Ile Thr Met Val Leu Ile Leu Pro Lys Pro
275 280 285
Glu Lys Ser Leu Ala Lys Val Glu Lys Glu Leu Thr Pro Glu Val Leu
290 295 300
Gln Glu Trp Leu Asp Glu Leu Glu Glu Met Met Leu Val Val His Met
305 310 315 320
Pro Arg Phe Arg Ile Glu Asp Gly Phe Ser Leu Lys Glu Gln Leu Gln
325 330 335
Asp Met Gly Leu Val Asp Leu Phe Ser Pro Glu Lys Ser Lys Leu Pro
340 345 350
Gly Ile Val Ala Glu Gly Arg Asp Asp Leu Tyr Val Ser Asp Ala Phe
355 360 365
His Lys Ala Phe Leu Glu Val Asn Glu Glu Gly Ser Glu Ala Ala Ala
370 375 380
Ser Thr Ala Val Val Arg Arg Val Arg Lys Glu Asn Pro Asn Arg Val
385 390 395 400
Thr Phe Lys Ala Asn Arg Pro Phe Leu Val Phe Ile Arg Glu Val Pro
405 410 415
Leu Asn Thr Ile Ile Phe Met Gly Arg Val Ala Asn Pro Cys Val Lys
420 425 430
<210>498
<211>6
<212>PRT
<213>Artificial Sequence
<220>
<223>replacement cleavage site RRVRKE
<400>498
Arg Arg Val Arg Lys Glu
1 5
<210>499
<211>432
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant AT3 sequence
<223>AT3/SLGRKI
<400>499
His Gly Ser Pro Val Asp Ile Cys Thr Ala Lys Pro Arg Asp Ile Pro
1 5 10 15
Met Asn Pro Met Cys Ile Tyr Arg Ser Pro Glu Lys Lys Ala Thr Glu
20 25 30
Asp Glu Gly Ser Glu Gln Lys Ile Pro Glu Ala Thr Asn Arg Arg Val
35 40 45
Trp Glu Leu Ser Lys Ala Asn Ser Arg Phe Ala Thr Thr Phe Tyr Gln
50 55 60
His Leu Ala Asp Ser Lys Asn Asp Asn Asp Asn Ile Phe Leu Ser Pro
65 70 75 80
Leu Ser Ile Ser Thr Ala Phe Ala Met Thr Lys Leu Gly Ala Cys Asn
85 90 95
Asp Thr Leu Gln Gln Leu Met Glu Val Phe Lys Phe Asp Thr Ile Ser
100 105 110
Glu Lys Thr Ser Asp Gln Ile His Phe Phe Phe Ala Lys Leu Asn Cys
115 120 125
Arg Leu Tyr Arg Lys Ala Asn Lys Ser Ser Lys Leu Val Ser Ala Asn
130 135 140
Arg Leu Phe Gly Asp Lys Ser Leu Thr Phe Asn Glu Thr Tyr Gln Asp
145 150 155 160
Ile Ser Glu Leu Val Tyr Gly Ala Lys Leu Gln Pro Leu Asp Phe Lys
165 170 175
Glu Asn Ala Glu Gln Ser Arg Ala Ala Ile Asn Lys Trp Val Ser Asn
180 185 190
Lys Thr Glu Gly Arg Ile Thr Asp Val Ile Pro Ser Glu Ala Ile Asn
195 200 205
Glu Leu Thr Val Leu Val Leu Val Asn Thr Ile Tyr Phe Lys Gly Leu
210 215 220
Trp Lys Ser Lys Phe Ser Pro Glu Asn Thr Arg Lys Glu Leu Phe Tyr
225 230 235 240
Lys Ala Asp Gly Glu Ser Cys Ser Ala Ser Met Met Tyr Gln Glu Gly
245 250 255
Lys Phe Arg Tyr Arg Arg Val Ala Glu Gly Thr Gln Val Leu Glu Leu
260 265 270
Pro Phe Lys Gly Asp Asp Ile Thr Met Val Leu Ile Leu Pro Lys Pro
275 280 285
Glu Lys Ser Leu Ala Lys Val Glu Lys Glu Leu Thr Pro Glu Val Leu
290 295 300
Gln Glu Trp Leu Asp Glu Leu Glu Glu Met Met Leu Val Val His Met
305 310 315 320
Pro Arg Phe Arg Ile Glu Asp Gly Phe Ser Leu Lys Glu Gln Leu Gln
325 330 335
Asp Met Gly Leu Val Asp Leu Phe Ser Pro Glu Lys Ser Lys Leu Pro
340 345 350
Gly Ile Val Ala Glu Gly Arg Asp Asp Leu Tyr Val Ser Asp Ala Phe
355 360 365
His Lys Ala Phe Leu Glu Val Asn Glu Glu Gly Ser Glu Ala Ala Ala
370 375 380
Ser Thr Ala Val Val Ser Leu Gly Arg Lys Ile Asn Pro Asn Arg Val
385 390 395 400
Thr Phe Lys Ala Asn Arg Pro Phe Leu Val Phe Ile Arg Glu Val Pro
405 410 415
Leu Asn Thr Ile Ile Phe Met Gly Arg Val Ala Asn Pro Cys Val Lys
420 425 430
<210>500
<211>432
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant AT3 sequence
<223>SKGRSL
<400>500
His Gly Ser Pro Val Asp Ile Cys Thr Ala Lys Pro Arg Asp Ile Pro
1 5 10 15
Met Asn Pro Met Cys Ile Tyr Arg Ser Pro Glu Lys Lys Ala Thr Glu
20 25 30
Asp Glu Gly Ser Glu Gln Lys Ile Pro Glu Ala Thr Asn Arg Arg Val
35 40 45
Trp Glu Leu Ser Lys Ala Asn Ser Arg Phe Ala Thr Thr Phe Tyr Gln
50 55 60
His Leu Ala Asp Ser Lys Asn Asp Asn Asp Asn Ile Phe Leu Ser Pro
65 70 75 80
Leu Ser Ile Ser Thr Ala Phe Ala Met Thr Lys Leu Gly Ala Cys Asn
85 90 95
Asp Thr Leu Gln Gln Leu Met Glu Val Phe Lys Phe Asp Thr Ile Ser
100 105 110
Glu Lys Thr Ser Asp Gln Ile His Phe Phe Phe Ala Lys Leu Asn Cys
115 120 125
Arg Leu Tyr Arg Lys Ala Asn Lys Ser Ser Lys Leu Val Ser Ala Asn
130 135 140
Arg Leu Phe Gly Asp Lys Ser Leu Thr Phe Asn Glu Thr Tyr Gln Asp
145 150 155 160
Ile Ser Glu Leu Val Tyr Gly Ala Lys Leu Gln Pro Leu Asp Phe Lys
165 170 175
Glu Asn Ala Glu Gln Ser Arg Ala Ala Ile Asn Lys Trp Val Ser Asn
180 185 190
Lys Thr Glu Gly Arg Ile Thr Asp Val Ile Pro Ser Glu Ala Ile Asn
195 200 205
Glu Leu Thr Val Leu Val Leu Val Asn Thr Ile Tyr Phe Lys Gly Leu
210 215 220
Trp Lys Ser Lys Phe Ser Pro Glu Asn Thr Arg Lys Glu Leu Phe Tyr
225 230 235 240
Lys Ala Asp Gly Glu Ser Cys Ser Ala Ser Met Met Tyr Gln Glu Gly
245 250 255
Lys Phe Arg Tyr Arg Arg Val Ala Glu Gly Thr Gln Val Leu Glu Leu
260 265 270
Pro Phe Lys Gly Asp Asp Ile Thr Met Val Leu Ile Leu Pro Lys Pro
275 280 285
Glu Lys Ser Leu Ala Lys Val Glu Lys Glu Leu Thr Pro Glu Val Leu
290 295 300
Gln Glu Trp Leu Asp Glu Leu Glu Glu Met Met Leu Val Val His Met
305 310 315 320
Pro Arg Phe Arg Ile Glu Asp Gly Phe Ser Leu Lys Glu Gln Leu Gln
325 330 335
Asp Met Gly Leu Val Asp Leu Phe Ser Pro Glu Lys Ser Lys Leu Pro
340 345 350
Gly Ile Val Ala Glu Gly Arg Asp Asp Leu Tyr Val Ser Asp Ala Phe
355 360 365
His Lys Ala Phe Leu Glu Val Asn Glu Glu Gly Ser Glu Ala Ala Ala
370 375 380
Ser Thr Ala Val Val Ser Lys Gly Arg Ser Leu Asn Pro Asn Arg Val
385 390 395 400
Thr Phe Lys Ala Asn Arg Pro Phe Leu Val Phe Ile Arg Glu Val Pro
405 410 415
Leu Asn Thr Ile Ile Phe Met Gly Arg Val Ala Asn Pro Cys Val Lys
420 425 430
<210>501
<211>6
<212>PRT
<213>Artificial Sequence
<220>
<223>replacement cleavage site SKGRSL
<400>501
Ser Lys Gly Arg Ser Leu
1 5
<210>502
<211>432
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant AT3 sequence
<223>AT3/PRFKII
<400>502
His Gly Ser Pro Val Asp Ile Cys Thr Ala Lys Pro Arg Asp Ile Pro
1 5 10 15
Met Asn Pro Met Cys Ile Tyr Arg Ser Pro Glu Lys Lys Ala Thr Glu
20 25 30
Asp Glu Gly Ser Glu Gln Lys Ile Pro Glu Ala Thr Asn Arg Arg Val
35 40 45
Trp Glu Leu Ser Lys Ala Asn Ser Arg Phe Ala Thr Thr Phe Tyr Gln
50 55 60
His Leu Ala Asp Ser Lys Asn Asp Asn Asp Asn Ile Phe Leu Ser Pro
65 70 75 80
Leu Ser Ile Ser Thr Ala Phe Ala Met Thr Lys Leu Gly Ala Cys Asn
85 90 95
Asp Thr Leu Gln Gln Leu Met Glu Val Phe Lys Phe Asp Thr Ile Ser
100 105 110
Glu Lys Thr Ser Asp Gln Ile His Phe Phe Phe Ala Lys Leu Asn Cys
115 120 125
Arg Leu Tyr Arg Lys Ala Asn Lys Ser Ser Lys Leu Val Ser Ala Asn
130 135 140
Arg Leu Phe Gly Asp Lys Ser Leu Thr Phe Asn Glu Thr Tyr Gln Asp
145 150 155 160
Ile Ser Glu Leu Val Tyr Gly Ala Lys Leu Gln Pro Leu Asp Phe Lys
165 170 175
Glu Asn Ala Glu Gln Ser Arg Ala Ala Ile Asn Lys Trp Val Ser Asn
180 185 190
Lys Thr Glu Gly Arg Ile Thr Asp Val Ile Pro Ser Glu Ala Ile Asn
195 200 205
Glu Leu Thr Val Leu Val Leu Val Asn Thr Ile Tyr Phe Lys Gly Leu
210 215 220
Trp Lys Ser Lys Phe Ser Pro Glu Asn Thr Arg Lys Glu Leu Phe Tyr
225 230 235 240
Lys Ala Asp Gly Glu Ser Cys Ser Ala Ser Met Met Tyr Gln Glu Gly
245 250 255
Lys Phe Arg Tyr Arg Arg Val Ala Glu Gly Thr Gln Val Leu Glu Leu
260 265 270
Pro Phe Lys Gly Asp Asp Ile Thr Met Val Leu Ile Leu Pro Lys Pro
275 280 285
Glu Lys Ser Leu Ala Lys Val Glu Lys Glu Leu Thr Pro Glu Val Leu
290 295 300
Gln Glu Trp Leu Asp Glu Leu Glu Glu Met Met Leu Val Val His Met
305 310 315 320
Pro Arg Phe Arg Ile Glu Asp Gly Phe Ser Leu Lys Glu Gln Leu Gln
325 330 335
Asp Met Gly Leu Val Asp Leu Phe Ser Pro Glu Lys Ser Lys Leu Pro
340 345 350
Gly Ile Val Ala Glu Gly Arg Asp Asp Leu Tyr Val Ser Asp Ala Phe
355 360 365
His Lys Ala Phe Leu Glu Val Asn Glu Glu Gly Ser Glu Ala Ala Ala
370 375 380
Ser Thr Ala Val Val Pro Arg Phe Lys Ile Ile Asn Pro Asn Arg Val
385 390 395 400
Thr Phe Lys Ala Asn Arg Pro Phe Leu Val Phe Ile Arg Glu Val Pro
405 410 415
Leu Asn Thr Ile Ile Phe Met Gly Arg Val Ala Asn Pro Cys Val Lys
420 425 430
<210>503
<211>6
<212>PRT
<213>Artificial Sequence
<220>
<223>replacement cleavage site PRFKII
<400>503
Pro Arg Phe Lys Ile Ile
1 5
<210>504
<211>723
<212>DNA
<213>Homo sapiens
<220>
<223>MT-SP1 protease domain
<400>504
gttgttgggg gcacggatgc ggatgagggc gagtggccct ggcaggtaag cctgcatgct 60
ctgggccagg gccacatctg cggtgcttcc ctcatctctc ccaactggct ggtctctgcc 120
gcacactgct acatcgatga cagaggattc aggtactcag accccacgca gtggacggcc 180
ttcctgggct tgcacgacca gagccagcgc agcgcccctg gggtgcagga gcgcaggctc 240
aagcgcatca tctcccaccc cttcttcaat gacttcacct tcgactatga catcgcgctg 300
ctggagctgg agaaaccggc agagtacagc tccatggtgc ggcccatctg cctgccggac 360
gcctcccatg tcttccctgc cggcaaggcc atctgggtca cgggctgggg acacacccag 420
tatggaggca ctggcgcgct gatcctgcaa aagggtgaga tccgcgtcat caaccagacc 480
acctgcgaga acctcctgcc gcagcagatc acgccgcgca tgatgtgcgt gggcttcctc 540
agcggcggcg tggactcctg ccagggtgat tccgggggac ccctgtccag cgtggaggcg 600
gatgggcgga tcttccaggc cggtgtggtg agctggggag acggctgcgc tcagaggaac 660
aagccaggcg tgtacacaag gctccctctg tttcgggact ggatcaaaga gaacactggg 720
gta 723
<210>505
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>MT-SP1 protease domain
<400>505
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Ash Thr Gly
225 230 235 240
Val
<210>506
<211>444
<212>PRT
<213>Homo sapiens
<220>
<223>MT-SP1 protease domain fusion
<223>STII/MT-SP1/GeneII
<400>506
Met Lys Lys Asn Ile Ala Phe Leu Leu Ala Ser Met Phe Val Phe Ser
1 5 10 15
Ile Ala Thr Asn Ala Tyr Ala Val Val Gly Gly Thr Asp Ala Asp Glu
20 25 30
Gly Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His
35 40 45
Ile Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala
50 55 60
His Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln
65 70 75 80
Trp Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro
85 90 95
Gly Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe
100 105 110
Asn Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys
115 120 125
Pro Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala
130 135 140
Ser His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly
145 150 155 160
His Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu
165 170 175
Ile Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln
180 185 190
Ile Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp
195 200 205
Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp
210 215 220
Gly Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala
225 230 235 240
Gln Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp
245 250 255
Trp Ile Lys Glu Asn Thr Gly Val Ser Gly Ser Ser Gly Gly Gly Ser
260 265 270
Glu Gly Gly Gly Ser Glu Gly Gly Gly Ser Glu Gly Gly Gly Ser Gly
275 280 285
Gly Gly Ser Gly Ser Gly Asp Phe Asp Tyr Glu Lys Met Ala Asn Ala
290 295 300
Asn Lys Gly Ala Met Thr Glu Asn Ala Asp Glu Asn Ala Leu Gln Ser
305 310 315 320
Asp Ala Lys Gly Lys Leu Asp Ser Val Ala Thr Asp Tyr Gly Ala Ala
325 330 335
Ile Asp Gly Phe Ile Gly Asp Val Ser Gly Leu Ala Asn Gly Asn Gly
340 345 350
Ala Thr Gly Asp Phe Ala Gly Ser Asn Ser Gln Met Ala Gln Val Gly
355 360 365
Asp Gly Asp Asn Ser Pro Leu Met Asn Asn Phe Arg Gln Tyr Leu Pro
370 375 380
Ser Leu Pro Gln Ser Val Glu Cys Arg Pro Phe Val Phe Ser Ala Gly
385 390 395 400
Lys Pro Tyr Glu Phe Ser Ile Asp Cys Asp Lys Ile Asn Leu Phe Arg
405 410 415
Gly Val Phe Ala Phe Leu Leu Tyr Val Ala Thr Phe Met Tyr Val Phe
420 425 430
Ser Thr Phe Ala Asn Ile Leu Arg Asn Lys Glu Ser
435 440
<210>507
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutated MT-SP1 protease domain
<223>CB496
<400>507
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>508
<211>44
<212>DNA
<213>Artificial Sequence
<220>
<223>MT-SP1 forward primer
<400>508
gcgcagatat cgtaccgcat atgaaaaaga atatcgcatt tctt 44
<210>509
<211>43
<212>DNA
<213>Artificial Sequence
<220>
<223>MT-SP1 reverse primer
<400>509
gtgcatgctg actgactgag ctcccgctta ccccagtgtt ctc 43
<210>510
<211>1335
<212>DNA
<213>Homo sapiens
<220>
<223>MT-SP1 protease domain fusion
<223>STII/MT-SP1/GeneIII
<400>510
atgaaaaaga atatcgcatt tcttcttgca tctatgttcg ttttttctat tgctacaaat 60
gcctatgcag ttgttggggg cacggatgcg gatgagggcg agtggccctg gcaggtaagc 120
ctgcatgctc tgggccaggg ccacatctgc ggtgcttccc tcatctctcc caactggctg 180
gtctctgccg cacactgcta catcgatgac agaggattca ggtactcaga ccccacgcag 240
tggacggcct tcctgggctt gcacgaccag agccagcgca gcgcccctgg ggtgcaggag 300
cgcaggctca agcgcatcat ctcccacccc ttcttcaatg acttcacctt cgactatgac 360
atcgcgctgc tggagctgga gaaaccggca gagtacagct ccatggtgcg gcccatctgc 420
ctgccggacg cctcccatgt cttccctgcc ggcaaggcca tctgggtcac gggctgggga 480
cacacccagt atggaggcac tggcgcgctg atcctgcaaa agggtgagat ccgcgtcatc 540
aaccagacca cctgcgagaa cctcctgccg cagcagatca cgccgcgcat gatgtgcgtg 600
ggcttcctca gcggcggcgt ggactcctgc cagggtgatt ccgggggacc cctgtccagc 660
gtggaggcgg atgggcggat cttccaggcc ggtgtggtga gctggggaga cggctgcgct 720
cagaggaaca agccaggcgt gtacacaagg ctccctctgt ttcgggactg gatcaaagag 780
aacactgggg taagcgggag ctccggtggt ggctctgagg gtggcggttc tgagggtggc 840
ggctctgagg gaggcggttc cggtggtggc tctggttccg gtgattttga ttatgaaaag 900
atggcaaacg ctaataaggg ggctatgacc gaaaatgccg atgaaaacgc gctacagtct 960
gacgctaaag gcaaacttga ttctgtcgct actgattacg gtgctgctat cgatggtttc 1020
attggtgacg tttccggcct tgctaatggt aatggtgcta ctggtgattt tgctggctct 1080
aattcccaaa tggctcaagt cggtgacggt gataattcac ctttaatgaa taatttccgt 1140
caatatttac cttccctccc tcaatcggtt gaatgtcgcc cttttgtctt tagcgctggt 1200
aaaccatatg aattttctat tgattgtgac aaaataaact tattccgtgg tgtctttgcg 1260
tttcttttat atgttgccac ctttatgtat gtattttcta cgtttgctaa catactgcgt 1320
aataaggagt cttaa 1335
<210>511
<211>23
<212>PRT
<213>Escherichia coli
<220>
<223>STII leader sequence
<400>511
Met Lys Lys Asn Ile Ala Phe Leu Leu Ala Ser Met Phe Val Phe Ser
1 5 10 15
Ile Ala Thr Asn Ala Tyr Ala
20
<210>512
<211>180
<212>PRT
<213>M13 phage
<220>
<223>GeneIII
<400>512
Ser Gly Ser Ser Gly Gly Gly Ser Glu Gly Gly Gly Ser Glu Gly Gly
1 5 10 15
Gly Ser Glu Gly Gly Gly Ser Gly Gly Gly Ser Gly Ser Gly Asp Phe
20 25 30
Asp Tyr Glu Lys Met Ala Asn Ala Asn Lys Gly Ala Met Thr Glu Asn
35 40 45
Ala Asp Glu Asn Ala Leu Gln Ser Asp Ala Lys Gly Lys Leu Asp Ser
50 55 60
Val Ala Thr Asp Tyr Gly Ala Ala Ile Asp Gly Phe Ile Gly Asp Val
65 70 75 80
Ser Gly Leu Ala Asn Gly Asn Gly Ala Thr Gly Asp Phe Ala Gly Ser
85 90 95
Asn Ser Gln Met Ala Gln Val Gly Asp Gly Asp Asn Ser Pro Leu Met
100 105 110
Asn Asn Phe Arg Gln Tyr Leu Pro Ser Leu Pro Gln Ser Val Glu Cys
115 120 125
Arg Pro Phe Val Phe Ser Ala Gly Lys Pro Tyr Glu Phe Ser Ile Asp
130 135 140
Cys Asp Lys Ile Asn Leu Phe Arg Gly Val Phe Ala Phe Leu Leu Tyr
145 150 155 160
Val Ala Thr Phe Met Tyr Val Phe Ser Thr Phe Ala Asn Ile Leu Arg
165 170 175
Asn Lys Glu Ser
180
<210>513
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site RQAR
<400>513
Arg G1n Ala Arg
1
<210>514
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>Synthesized protease cleavage site SLGR
<400>514
Ser Leu Gly Arg
1
<210>515
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CB469
<400>515
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>516
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP-1 protease domain
<223>CPC-0019595
<400>516
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Thr Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asp Gln Ala
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Leu Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Val Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>517
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0023085
<400>517
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala lle Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>518
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0024153
<400>518
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Thr Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Gly
<210>519
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0025366
<400>519
Val Val Gly Gly Thr Asp Ala Glu Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Pro Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Ile Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>520
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0025387
<400>520
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>521
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0025582
<400>521
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Phe Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Gly
<210>522
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP-1 protease domain
<223>CPC-0025720
<400>522
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly Arg Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Arg Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>523
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0025876
<400>523
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Phe Leu Pro Gln Gln Ile Thr Pro Arg Mer Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>524
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0025974
<400>524
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>525
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0026100
<400>525
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Val Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Gly
<210>526
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0026323
<400>526
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Met Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Asp Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Leu Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>527
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0027399
<400>527
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Leu Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>528
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0027706
<400>528
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Tyr Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Val Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys His Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ile Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>529
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0027797
<400>529
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
Arg Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Ser Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Val Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>530
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0028017
<400>530
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Val Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>531
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0028333
<400>531
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Lys Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Leu Phe Asn Asp Tyr Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Val Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>532
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0028341
<400>532
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Thr Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Ser Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Leu Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>533
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0033634
<400>533
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Leu Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Leu Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>534
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0028791
<400>534
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>535
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0027484
<400>535
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Lys Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>536
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 protease domain
<223>CPC-0028993
<400>536
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Ser Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Ser Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Arg Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>537
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0019595
<400>537
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Thr Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asp Gln Ala Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Leu Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Val Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>538
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0023085
<400>538
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser C s Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>539
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0024153
<400>539
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Ar Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Thr Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Gly
850 855
<210>540
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0025366
<400>540
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Glu Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Pro Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Ile Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>541
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0025387
<400>541
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>542
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0025582
<400>542
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Phe Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Gly
850 855
<210>543
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0025720
<400>543
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly Arg
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Arg Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>544
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0025876
<400>544
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Phe Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>545
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0025974
<400>545
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>546
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0026100
<400>546
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Val Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Gly
850 855
<210>547
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0026323
<400>547
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Met Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Asp Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Leu
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>548
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-002739
<400>548
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Leu Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>549
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0027706
<400>549
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Ar Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Tyr Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Val Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys His Gly Asp Ser Gly Gly Pro Leu Ser Ile Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>550
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0027797
<400>550
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu Arg Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Ser Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Val Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>551
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0028017
<400>551
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Gys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Val Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>552
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0028333
<400>552
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Lys Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Leu Phe Asn
690 695 700
Asp Tyr Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Val Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>553
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0028341
<400>553
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Thr
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Ser Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Leu Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>554
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0033634
<400>554
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Leu Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Leu Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>555
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0028791
<400>555
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>556
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0027484
<400>556
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Lys
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>557
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MT-SP1 full length
<223>CPC-0028993
<400>557
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Ser Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Ser Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Arg Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>558
<211>26
<212>PRT
<213>Artificial Sequence
<220>
<223>N-terminal sequence of pIVEX 2.3d MTSP
<400>558
Met Glu Lys Thr Arg His His His His His His Ser Gly Ser Asp Cys
1 5 10 15
Gly Leu Arg Ser Phe Thr Arg Gln Ala Arg
20 25
<210>559
<211>3560
<212>DNA
<213>Artificial Sequence
<220>
<223>pIVEX 2.3d vector
<400>559
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60
cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120
ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180
accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 240
ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 300
attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 360
gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgcat gcaaggagat 420
ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct 480
catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc 540
agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga 600
gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct 660
agaaataatt ttgtttaact ttaagaagga gatataccat ggcacatatg agcggccgcg 720
tcgactcgag cgagctcccg gggggggttc tcatcatcat catcatcatt aataaaaggg 780
cgaattccag cacactggcg gccgttacta gtggatccgg ctgctaacaa agcccgaaag 840
gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct 900
aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata tccacaggac 960
gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag cgagcaggac 1020
tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata gaaattgcat 1080
caacgcatat agcgctagca gcacgccata gtgactggcg atgctgtcgg aatggacgat 1140
atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt 1200
gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg 1260
ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac 1320
atgagaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 1380
tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 1440
ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 1500
ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 1560
ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 1620
agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 1680
catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 1740
tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 1800
gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 1860
ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 1920
cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 1980
caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 2040
ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 2100
taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 2160
taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 2220
cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 2280
tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 2340
gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 2400
catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 2460
atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 2520
ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 2580
gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 2640
agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 2700
gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 2760
agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 2820
catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 2880
aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 2940
gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 3000
taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 3060
caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 3120
ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 3180
ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 3240
tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 3300
aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 3360
actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 3420
catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 3480
agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 3540
tatcacgagg ccctttcgtc 3560
<210>560
<211>36
<212>DNA
<213>Artificial Sequence
<220>
<223>pQE-Insert-F2 forward primer
<400>560
ttcacgagac aggctcgtgt tgttgggggc acggat 36
<210>561
<211>37
<212>DNA
<213>Artificial Sequence
<220>
<223>pQE-Insert-R3 reverse primer
<400>561
cagctaatta agcttattat accccagtgt tctcttt 37
<210>562
<211>30
<212>DNA
<213>Artificial Sequence
<220>
<223>pQE-Linear-F2 primer
<400>562
acgagcctgt ctcgtgaatg accgcagccc 30
<210>563
<211>31
<212>DNA
<213>Artificial Sequence
<220>
<223>pQE-Linear-R1 primer
<400>563
taataagctt aattagctga gcttggactc c 31
<210>564
<211>19
<212>DNA
<213>Artificial Sequence
<220>
<223>forward primer：MTSP-5F
<400>564
ggagaaaccg gcagagtac 19
<210>565
<211>19
<212>DNA
<213>Artificial Sequence
<220>
<223>reverse primer MTSP-5R
<400>565
ggttctcgca ggtggtctg 19
<210>566
<211>5443
<212>DNA
<213>Artificial Sequence
<220>
<223>pET21a vector
<400>566
atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 60
ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 120
tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt 180
cgacggagct cgaattcgga tccgcgaccc atttgctgtc caccagtcat gctagccata 240
tgtatatctc cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc 300
acaattcccc tatagtgagt cgtattaatt tcgcgggatc gagatctcga tcctctacgc 360
cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc 420
cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg 480
cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc 540
accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat 600
gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct 660
ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac 720
cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg 780
tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg 840
cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc 900
tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga 960
ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg 1020
gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga 1080
tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg 1140
ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc 1200
atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg 1260
cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata 1320
aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca 1380
tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc 1440
tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc 1500
gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata 1560
tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc 1620
gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac 1680
tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg 1740
ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc 1800
aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg 1860
agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca 1920
cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc 1980
attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta 2040
ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc 2100
ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc 2160
ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca 2220
ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca 2280
acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc 2340
tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct 2400
acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc 2460
atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca 2520
gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga 2580
aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc 2640
cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat 2700
gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc 2760
ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 2820
acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 2880
gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact 2940
ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga 3000
aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct 3060
cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 3120
ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 3180
ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 3240
cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 3300
actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 3360
cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 3420
tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 3480
gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 3540
caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 3600
agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 3660
tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 3720
tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 3780
gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 3840
gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 3900
aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 3960
atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 4020
gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 4080
acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 4140
ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 4200
tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 4260
ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg 4320
ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 4380
atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 4440
taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 4500
catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 4560
atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 4620
acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 4680
aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 4740
ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 4800
cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 4860
atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 4920
ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat 4980
tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt 5040
taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg 5100
gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt 5160
caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc 5220
aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg 5280
atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa 5340
aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc 5400
cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca 5443
<210>567
<211>379
<212>PRT
<213>Homo sapiens
<220>
<223>PAI-1 stable
<400>567
Val His His Pro Pro Ser Tyr Val Ala His Leu Ala Ser Asp Phe Gly
1 5 10 15
Val Arg Val Phe Gln Gln Val Ala Gln Ala Ser Lys Asp Arg Asn Val
20 25 30
Val Phe Ser Pro Tyr Gly Val Ala Ser Val Leu Ala Met Leu Gln Leu
35 40 45
Thr Thr Gly Gly Glu Thr Gln Gln Gln Ile Gln Ala Ala Met Gly Phe
50 55 60
Lys Ile Asp Asp Lys Gly Met Ala Pro Ala Leu Arg His Leu Tyr Lys
65 70 75 80
Glu Leu Met Gly Pro Trp Asn Lys Asp Glu Ile Ser Thr Thr Asp Ala
85 90 95
Ile Phe Val Gln Arg Asp Leu Lys Leu Val Gln Gly Phe Met Pro His
100 105 110
Phe Phe Arg Leu Phe Arg Ser Thr Val Lys Gln Val Asp Phe Ser Glu
115 120 125
Val Glu Arg Ala Arg Phe Ile Ile Asn Asp Trp Val Lys Thr His Thr
130 135 140
Lys Gly Met Ile Ser His Leu Leu Gly Thr Gly Ala Val Asp Gln Leu
145 150 155 160
Thr Arg Leu Val Leu Val Asn Ala Leu Tyr Phe Asn Gly Gln Trp Lys
165 170 175
Thr Pro Phe Pro Asp Ser Ser Thr His Arg Arg Leu Phe His Lys Ser
180 185 190
Asp Gly Ser Thr Val Ser Val Pro Met Met Ala Gln Thr Asn Lys Phe
195 200 205
Asn Tyr Thr Glu Phe Thr Thr Pro Asp Gly His Tyr Tyr Asp Ile Leu
210 215 220
Glu Leu Pro Tyr His Gly Asp Thr Leu Ser Met Phe Ile Ala Ala Pro
225 230 235 240
Tyr Glu Lys Glu Val Pro Leu Ser Ala Leu Thr Asn Ile Leu Ser Ala
245 250 255
Gln Leu Ile Ser His Trp Lys Gly Asn Met Thr Arg Leu Pro Arg Leu
260 265 270
Leu Val Leu Pro Lys Phe Ser Leu Glu Thr Glu Val Asp Leu Arg Lys
275 280 285
Pro Leu Glu Asn Leu Gly Met Thr Asp Met Phe Arg Gln Phe Gln Ala
290 295 300
Asp Phe Thr Ser Leu Ser Asp Gln Glu Pro Leu His Val Ala Leu Ala
305 310 315 320
Leu Gln Lys Val Lys Ile Glu Val Asn Glu Ser Gly Thr Val Ala Ser
325 330 335
Ser Ser Thr Ala Val Ile Val Ser Ala Arg Met Ala Pro Glu Glu Ile
340 345 350
Ile Ile Asp Arg Pro Phe Leu Phe Val Val Arg His Asn Pro Thr Gly
355 360 365
Thr Val Leu Phe Met Gly Gln Val Met Glu Pro
370 375
<210>568
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>I136T/N164D/T166A/F184(A)L/D217V
<400>568
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Thr Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asp Gln Ala
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Leu Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Val Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>569
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>I41F
<400>569
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>570
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>I41F/A126T/V244G
<400>570
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Thr Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Gly
<210>571
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>D23E/I41F/T98P/T144I
<400>571
Val Val Gly Gly Thr Asp Ala Glu Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Pro Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Ile Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>572
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>I41F
<400>572
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>573
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>I41F/L171F/V244G
<400>573
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Phe Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Gly
<210>574
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>H143R/Q175R
<400>574
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly Arg Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Arg Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Va1 Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>575
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>I41F/L171F
<400>575
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Phe Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>576
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>R230W
<400>576
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>577
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>I41F/I154V/V244G
<400>577
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Val Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Gly
<210>578
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>I141F/L52M/V129D/Q221(A)L
<400>578
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Phe Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Met Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Asp Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Leu Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>579
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>F99L
<400>579
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Leu Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>580
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>F97Y/I136V/Q192H/S201I
<400>580
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Tyr Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Val Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys His Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ile Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>581
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>H71R/P131S/D217V
<400>581
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
Arg Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Ser Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Val Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>582
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>D217V
<400>582
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Val Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>583
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>T65K/F93L/F97Y/D217V
<400>583
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Lys Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Leu Phe Asn Asp Tyr Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Val Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>584
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>I41T/P173S/Q209L
<400>584
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Thr Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Ser Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Leu Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>585
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>F97L//F234L
<400>585
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Leu Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Leu Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>586
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>Q175R
<400>586
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>587
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 protease domain
<223>N95K
<400>587
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Lys Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>588
<211>241
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1protease domain
<223>Y60(G)S
<400>588
Val Val Gly Gly Thr Asp Ala Asp Glu Gly Glu Trp Pro Trp Gln Val
1 5 10 15
Ser Leu His Ala Leu Gly Gln Gly His Ile Cys Gly Ala Ser Leu Ile
20 25 30
Ser Pro Asn Trp Leu Val Ser Ala Ala His Cys Tyr Ile Asp Asp Arg
35 40 45
Gly Phe Arg Ser Ser Asp Pro Thr Gln Trp Thr Ala Phe Leu Gly Leu
50 55 60
His Asp Gln Ser Gln Arg Ser Ala Pro Gly Val Gln Glu Arg Arg Leu
65 70 75 80
Lys Arg Ile Ile Ser His Pro Phe Phe Asn Asp Phe Thr Phe Asp Tyr
85 90 95
Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro Ala Glu Tyr Ser Ser Met
100 105 110
Val Arg Pro Ile Cys Leu Pro Asp Ala Ser His Val Phe Pro Ala Gly
115 120 125
Lys Ala Ile Trp Val Thr Gly Trp Gly His Thr Gln Tyr Gly Gly Thr
130 135 140
Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile Arg Val Ile Asn Gln Thr
145 150 155 160
Thr Cys Glu Asn Leu Leu Pro Gln Arg Ile Thr Pro Arg Met Met Cys
165 170 175
Val Gly Phe Leu Ser Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly
180 185 190
Gly Pro Leu Ser Ser Val Glu Ala Asp Gly Arg Ile Phe Gln Ala Gly
195 200 205
Val Val Ser Trp Gly Asp Gly Cys Ala Gln Arg Asn Lys Pro Gly Val
210 215 220
Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp Ile Lys Glu Asn Thr Gly
225 230 235 240
Val
<210>589
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>I136T/N164D/T166A/F184(A)L/D217V
<400>589
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile As Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Thr Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asp Gln Ala Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Leu Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Val Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>590
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>I41F
<400>590
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>591
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>I41F/A126T/V244G
<400>591
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Thr Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Gly
850 855
<210>592
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>D23E/I41F/T98P/T144I
<400>592
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Glu Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Pro Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Ile Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>593
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>I41F
<400>593
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>594
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>I41F/L171F/V244G
<400>594
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Phe Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Gly
850 855
<210>595
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>H143R/Q175R
<400>595
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Sar Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly Arg
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Arg Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>596
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>I41F/L171F
<400>596
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
l00 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile As Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Phe Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>597
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>R230W
<400>597
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>598
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>I41F/I154V/V244G
<400>598
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Iie Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Val Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Gly
850 855
<210>599
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>I141F/L52M/V129D/Q221(A)L
<400>599
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
695 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Phe
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Met Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 580 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Asp Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Leu
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>600
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>F99L
<400>600
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Tr Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Leu Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>601
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>F97Y/I136V/Q192H/S201I
<400>601
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trn
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
695 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Tyr Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Val Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys His Gly Asp Ser Gly Gly Pro Leu Ser Ile Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>602
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>H71R/P131S/D217V
<400>602
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu Arg Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Ser Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Val Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>603
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>D217V
<400>603
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Val Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>604
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>T65K/F93L/F97Y/D217V
<400>604
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Lys Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Leu Phe Asn
690 695 700
Asp Tyr Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Val Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>605
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>I41T/P173S/Q209L
<400>605
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Thr
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Ser Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Leu Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>606
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>F97L//F234L
<400>606
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Leu Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Leu Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>607
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>Q175R
<400>607
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>608
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>N95K
<400>608
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Tyr Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Lys
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Gln Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>609
<211>855
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant MTSP1 full length
<223>Y60(G)S
<400>609
Met Gly Ser Asp Arg Ala Arg Lys Gly Gly Gly Gly Pro Lys Asp Phe
1 5 10 15
Gly Ala Gly Leu Lys Tyr Asn Ser Arg His Glu Lys Val Asn Gly Leu
20 25 30
Glu Glu Gly Val Glu Phe Leu Pro Val Asn Asn Val Lys Lys Val Glu
35 40 45
Lys His Gly Pro Gly Arg Trp Val Val Leu Ala Ala Val Leu Ile Gly
50 55 60
Leu Leu Leu Val Leu Leu Gly Ile Gly Phe Leu Val Trp His Leu Gln
65 70 75 80
Tyr Arg Asp Val Arg Val Gln Lys Val Phe Asn Gly Tyr Met Arg Ile
85 90 95
Thr Asn Glu Asn Phe Val Asp Ala Tyr Glu Asn Ser Asn Ser Thr Glu
100 105 110
Phe Val Ser Leu Ala Ser Lys Val Lys Asp Ala Leu Lys Leu Leu Tyr
115 120 125
Ser Gly Val Pro Phe Leu Gly Pro Tyr His Lys Glu Ser Ala Val Thr
130 135 140
Ala Phe Ser Glu Gly Ser Val Ile Ala Tyr Tyr Trp Ser Glu Phe Ser
145 150 155 160
Ile Pro Gln His Leu Val Glu Glu Ala Glu Arg Val Met Ala Glu Glu
165 170 175
Arg Val Val Met Leu Pro Pro Arg Ala Arg Ser Leu Lys Ser Phe Val
180 185 190
Val Thr Ser Val Val Ala Phe Pro Thr Asp Ser Lys Thr Val Gln Arg
195 200 205
Thr Gln Asp Asn Ser Cys Ser Phe Gly Leu His Ala Arg Gly Val Glu
210 215 220
Leu Met Arg Phe Thr Thr Pro Gly Phe Pro Asp Ser Pro Tyr Pro Ala
225 230 235 240
His Ala Arg Cys Gln Trp Ala Leu Arg Gly Asp Ala Asp Ser Val Leu
245 250 255
Ser Leu Thr Phe Arg Ser Phe Asp Leu Ala Ser Cys Asp Glu Arg Gly
260 265 270
Ser Asp Leu Val Thr Val Tyr Asn Thr Leu Ser Pro Met Glu Pro His
275 280 285
Ala Leu Val Gln Leu Cys Gly Thr Tyr Pro Pro Ser Tyr Asn Leu Thr
290 295 300
Phe His Ser Ser Gln Asn Val Leu Leu Ile Thr Leu Ile Thr Asn Thr
305 310 315 320
Glu Arg Arg His Pro Gly Phe Glu Ala Thr Phe Phe Gln Leu Pro Arg
325 330 335
Met Ser Ser Cys Gly Gly Arg Leu Arg Lys Ala Gln Gly Thr Phe Asn
340 345 350
Ser Pro Tyr Tyr Pro Gly His Tyr Pro Pro Asn Ile Asp Cys Thr Trp
355 360 365
Asn Ile Glu Val Pro Asn Asn Gln His Val Lys Val Arg Phe Lys Phe
370 375 380
Phe Tyr Leu Leu Glu Pro Gly Val Pro Ala Gly Thr Cys Pro Lys Asp
385 390 395 400
Tyr Val Glu Ile Asn Gly Glu Lys Tyr Cys Gly Glu Arg Ser Gln Phe
405 410 415
Val Val Thr Ser Asn Ser Asn Lys Ile Thr Val Arg Phe His Ser Asp
420 425 430
Gln Ser Tyr Thr Asp Thr Gly Phe Leu Ala Glu Tyr Leu Ser Tyr Asp
435 440 445
Ser Ser Asp Pro Cys Pro Gly Gln Phe Thr Cys Arg Thr Gly Arg Cys
450 455 460
Ile Arg Lys Glu Leu Arg Cys Asp Gly Trp Ala Asp Cys Thr Asp His
465 470 475 480
Ser Asp Glu Leu Asn Cys Ser Cys Asp Ala Gly His Gln Phe Thr Cys
485 490 495
Lys Asn Lys Phe Cys Lys Pro Leu Phe Trp Val Cys Asp Ser Val Asn
500 505 510
Asp Cys Gly Asp Asn Ser Asp Glu Gln Gly Cys Ser Cys Pro Ala Gln
515 520 525
Thr Phe Arg Cys Ser Asn Gly Lys Cys Leu Ser Lys Ser Gln Gln Cys
530 535 540
Asn Gly Lys Asp Asp Cys Gly Asp Gly Ser Asp Glu Ala Ser Cys Pro
545 550 555 560
Lys Val Asn Val Val Thr Cys Thr Lys His Thr Tyr Arg Cys Leu Asn
565 570 575
Gly Leu Cys Leu Ser Lys Gly Asn Pro Glu Cys Asp Gly Lys Glu Asp
580 585 590
Cys Ser Asp Gly Ser Asp Glu Lys Asp Cys Asp Cys Gly Leu Arg Ser
595 600 605
Phe Thr Arg Gln Ala Arg Val Val Gly Gly Thr Asp Ala Asp Glu Gly
610 615 620
Glu Trp Pro Trp Gln Val Ser Leu His Ala Leu Gly Gln Gly His Ile
625 630 635 640
Cys Gly Ala Ser Leu Ile Ser Pro Asn Trp Leu Val Ser Ala Ala His
645 650 655
Cys Tyr Ile Asp Asp Arg Gly Phe Arg Ser Ser Asp Pro Thr Gln Trp
660 665 670
Thr Ala Phe Leu Gly Leu His Asp Gln Ser Gln Arg Ser Ala Pro Gly
675 680 685
Val Gln Glu Arg Arg Leu Lys Arg Ile Ile Ser His Pro Phe Phe Asn
690 695 700
Asp Phe Thr Phe Asp Tyr Asp Ile Ala Leu Leu Glu Leu Glu Lys Pro
705 710 715 720
Ala Glu Tyr Ser Ser Met Val Arg Pro Ile Cys Leu Pro Asp Ala Ser
725 730 735
His Val Phe Pro Ala Gly Lys Ala Ile Trp Val Thr Gly Trp Gly His
740 745 750
Thr Gln Tyr Gly Gly Thr Gly Ala Leu Ile Leu Gln Lys Gly Glu Ile
755 760 765
Arg Val Ile Asn Gln Thr Thr Cys Glu Asn Leu Leu Pro Gln Arg Ile
770 775 780
Thr Pro Arg Met Met Cys Val Gly Phe Leu Ser Gly Gly Val Asp Ser
785 790 795 800
Cys Gln Gly Asp Ser Gly Gly Pro Leu Ser Ser Val Glu Ala Asp Gly
805 810 815
Arg Ile Phe Gln Ala Gly Val Val Ser Trp Gly Asp Gly Cys Ala Gln
820 825 830
Arg Asn Lys Pro Gly Val Tyr Thr Arg Leu Pro Leu Phe Arg Asp Trp
835 840 845
Ile Lys Glu Asn Thr Gly Val
850 855
<210>610
<211>380
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant mature PAI-1
<223>PAI-1/RRAR
<400>610
Met Val His His Pro Pro Ser Tyr Val Ala His Leu Ala Ser Asp Phe
1 5 10 15
Gly Val Arg Val Phe Gln Gln Val Ala Gln Ala Ser Lys Asp Arg Asn
20 25 30
Val Val Phe Ser Pro Tyr Gly Val Ala Ser Val Leu Ala Met Leu Gln
35 40 45
Leu Thr Thr Gly Gly Glu Thr Gln Gln Gln Ile Gln Ala Ala Met Gly
50 55 60
Phe Lys Ile Asp Asp Lys Gly Met Ala Pro Ala Leu Arg His Leu Tyr
65 70 75 80
Lys Glu Leu Met Gly Pro Trp Asn Lys Asp Glu Ile Ser Thr Thr Asp
85 90 95
Ala Ile Phe Val Gln Arg Asp Leu Lys Leu Val Gln Gly Phe Met Pro
100 105 110
His Phe Phe Arg Leu Phe Arg Ser Thr Val Lys Gln Val Asp Phe Ser
115 120 125
Glu Val Glu Arg Ala Arg Phe Ile Ile Asn Asp Trp Val Lys Thr His
130 135 140
Thr Lys Gly Met Ile Ser Asn Leu Leu Gly Lys Gly Ala Val Asp Gln
145 150 155 160
Leu Thr Arg Leu Val Leu Val Asn Ala Leu Tyr Phe Asn Gly Gln Trp
165 170 175
Lys Thr Pro Phe Pro Asp Ser Ser Thr His Arg Arg Leu Phe His Lys
180 185 190
Ser Asp Gly Ser Thr Val Ser Val Pro Met Met Ala Gln Thr Asn Lys
195 200 205
Phe Asn Tyr Thr Glu Phe Thr Thr Pro Asp Gly His Tyr Tyr Asp Ile
210 215 220
Leu Glu Leu Pro Tyr His Gly Asp Thr Leu Ser Met Phe Ile Ala Ala
225 230 235 240
Pro Tyr Glu Lys Glu Val Pro Leu Ser Ala Leu Thr Asn Ile Leu Ser
245 250 255
Ala Gln Leu Ile Ser His Trp Lys Gly Asn Met Thr Arg Leu Pro Arg
260 265 270
Leu Leu Val Leu Pro Lys Phe Ser Leu Glu Thr Glu Val Asp Leu Arg
275 280 285
Lys Pro Leu Glu Asn Leu Gly Met Thr Asp Met Phe Arg Gln Phe Gln
290 295 300
Ala Asp Phe Thr Ser Leu Ser Asp Gln Glu Pro Leu His Val Ala Gln
305 310 315 320
Ala Leu Gln Lys Val Lys Ile Glu Val Asn Glu Ser Gly Thr Val Ala
325 330 335
Ser Ser Ser Thr Ala Val Ile Arg Arg Ala Arg Met Ala Pro Glu Glu
340 345 350
Ile Ile Met Asp Arg Pro Phe Leu Phe Val Val Arg His Asn Pro Thr
355 360 365
Gly Thr Val Leu Phe Met Gly Gln Val Met Glu Pro
370 375 380
<210>611
<211>380
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant mature PAI-1
<223>PAI-1/69(containing PFGRS)
<400>611
Met Val His His Pro Pro Ser Tyr Val Ala His Leu Ala Ser Asp Phe
1 5 10 15
Gly Val Arg Val Phe Gln Gln Val Ala Gln Ala Ser Lys Asp Arg Asn
20 25 30
Val Val Phe Ser Pro Tyr Gly Val Ala Ser Val Leu Ala Met Leu Gln
35 40 45
Leu Thr Thr Gly Gly Glu Thr Gln Gln Gln Ile Gln Ala Ala Met Gly
50 55 60
Phe Lys Ile Asp Asp Lys Gly Met Ala Pro Ala Leu Arg His Leu Tyr
65 70 75 80
Lys Glu Leu Met Gly Pro Trp Asn Lys Asp Glu Ile Ser Thr Thr Asp
85 90 95
Ala Ile Phe Val Gln Arg Asp Leu Lys Leu Val Gln Gly Phe Met Pro
100 105 110
His Phe Phe Arg Leu Phe Arg Ser Thr Val Lys Gln Val Asp Phe Ser
115 120 125
Glu Val Glu Arg Ala Arg Phe Ile Ile Asn Asp Trp Val Lys Thr His
130 135 140
Thr Lys Gly Met Ile Ser Asn Leu Leu Gly Lys Gly Ala Val Asp Gln
145 150 155 160
Leu Thr Arg Leu Val Leu Val Asn Ala Leu Tyr Phe Asn Gly Gln Trp
165 170 175
Lys Thr Pro Phe Pro Asp Ser Ser Thr His Arg Arg Leu Phe His Lys
180 185 190
Ser Asp Gly Ser Thr Val Ser Val Pro Met Met Ala Gln Thr Asn Lys
195 200 205
Phe Asn Tyr Thr Glu Phe Thr Thr Pro Asp Gly His Tyr Tyr Asp Ile
210 215 220
Leu Glu Leu Pro Tyr His Gly Asp Thr Leu Ser Met Phe Ile Ala Ala
225 230 235 240
Pro Tyr Glu Lys Glu Val Pro Leu Ser Ala Leu Thr Asn Ile Leu Ser
245 250 255
Ala Gln Leu Ile Ser His Trp Lys Gly Asn Met Thr Arg Leu Pro Arg
260 265 270
Leu Leu Val Leu Pro Lys Phe Ser Leu Glu Thr Glu Val Asp Leu Arg
275 280 285
Lys Pro Leu Glu Asn Leu Gly Met Thr Asp Met Phe Arg Gln Phe Gln
290 295 300
Ala Asp Phe Thr Ser Leu Ser Asp Gln Glu Pro Leu His Val Ala Gln
305 310 315 320
Ala Leu Gln Lys Val Lys Ile Glu Val Asn Glu Ser Gly Thr Val Ala
325 330 335
Ser Ser Ser Thr Ala Val Ile Pro Phe Gly Arg Ser Ala Pro Glu Glu
340 345 350
Ile Ile Met Asp Arg Pro Phe Leu Phe Val Val Arg His Asn Pro Thr
355 360 365
Gly Thr Val Leu Phe Met Gly Gln Val Met Glu Pro
370 375 380
<210>612
<211>1395
<212>DNA
<213>Homo sapiens
<220>
<223>ATIII
<400>612
atgtattcca atgtgatagg aactgtaacc tctggaaaaa ggaaggttta tcttttgtcc 60
ttgctgctca ttggcttctg ggactgcgtg acctgtcacg ggagccctgt ggacatctgc 120
acagccaagc cgcgggacat tcccatgaat cccatgtgca tttaccgctc cccggagaag 180
aaggcaactg aggatgaggg ctcagaacag aagatcccgg aggccaccaa ccggcgtgtc 240
tgggaactgt ccaaggccaa ttcccgcttt gctaccactt tctatcagca cctggcagat 300
tccaagaatg acaatgataa cattttcctg tcacccctga gtatctccac ggcttttgct 360
atgaccaagc tgggtgcctg taatgacacc ctccagcaac tgatggaggt atttaagttt 420
gacaccatat ctgagaaaac atctgatcag atccacttct tctttgccaa actgaactgc 480
cgactctatc gaaaagccaa caaatcctcc aagttagtat cagccaatcg cctttttgga 540
gacaaatccc ttaccttcaa tgagacctac caggacatca gtgagttggt atatggagcc 600
aagctccagc ccctggactt caaggaaaat gcagagcaat ccagagcggc catcaacaaa 660
tgggtgtcca ataagaccga aggccgaatc accgatgtca ttccctcgga agccatcaat 720
gagctcactg ttctggtgct ggttaacacc atttacttca agggcctgtg gaagtcaaag 780
ttcagccctg agaacacaag gaaggaactg ttctacaagg ctgatggaga gtcgtgttca 840
gcatctatga tgtaccagga aggcaagttc cgttatcggc gcgtggctga aggcacccag 900
gtgcttgagt tgcccttcaa aggtgatgac atcaccatgg tcctcatctt gcccaagcct 960
gagaagagcc tggccaaggt ggagaaggaa ctcaccccag aggtgctgca ggagtggctg 1020
gatgaattgg aggagatgat gctggtggtc cacatgcccc gcttccgcat tgaggacggc 1080
ttcagtttga aggagcagct gcaagacatg ggccttgtcg atctgttcag ccctgaaaag 1140
tccaaactcc caggtattgt tgcagaaggc cgagatgacc tctatgtctc agatgcattc 1200
cataaggcat ttcttgaggt aaatgaagaa ggcagtgaag cagctgcaag taccgctgtt 1260
gtgattgctg gccgttcgct aaaccccaac agggtgactt tcaaggccaa caggcctttc 1320
ctggttttta taagagaagt tcctctgaac actattatct tcatgggcag agtagccaac 1380
ccttgtgtta agtaa 1395
<210>613
<211>1458
<212>DNA
<213>Artificial Sequence
<220>
<223>AT3 His tag fusion protein
<400>613
atgctactag taaatcagtc acaccaaggc ttcaataagg aacacacaag caagatggta 60
agcgctattg ttttatatgt gcttttggcg gcggcggcgc attctgcctt tgcggcggat 120
cttggatccc acgggagccc tgtggacatc tgcacagcca agccgcggga cattcccatg 180
aatcccatgt gcatttaccg ctccccggag aagaaggcaa ctgaggatga gggctcagaa 240
cagaagatcc cggaggccac caaccggcgt gtctgggaac tgtccaaggc caattcccgc 300
tttgctacca ctttctatca gcacctggca gattccaaga atgacaatga taacattttc 360
ctgtcacccc tgagtatctc cacggctttt gctatgacca agctgggtgc ctgtaatgac 420
accctccagc aactgatgga ggtatttaag tttgacacca tatctgagaa aacatctgat 480
cagatccact tcttctttgc caaactgaac tgccgactct atcgaaaagc caacaaatcc 540
tccaagttag tatcagccaa tcgccttttt ggagacaaat cccttacctt caatgagacc 600
taccaggaca tcagtgagtt ggtatatgga gccaagctcc agcccctgga cttcaaggaa 660
aatgcagagc aatccagagc ggccatcaac aaatgggtgt ccaataagac cgaaggccga 720
atcaccgatg tcattccctc ggaagccatc aatgagctca ctgttctggt gctggttaac 780
accatttact tcaagggcct gtggaagtca aagttcagcc ctgagaacac aaggaaggaa 840
ctgttctaca aggctgatgg agagtcgtgt tcagcatcta tgatgtacca ggaaggcaag 900
ttccgttatc ggcgcgtggc tgaaggcacc caggtgcttg agttgccctt caaaggtgat 960
gacatcacca tggtcctcat cttgcccaag cctgagaaga gcctggccaa ggtagagaag 1020
gaactcaccc cagaggtgct gcaagagtgg ctggatgaat tggaggagat gatgctggtg 1080
gtccacatgc cccgcttccg cattgaggac ggcttcagtt tgaaggagca gctgcaagac 1140
atgggccttg tcgatctgtt cagccctgaa aagtccaaac tcccaggtat tgttgcagaa 1200
ggccgagatg acctctatgt ctcagatgca ttccataagg catttcttga ggtaaatgaa 1260
gaaggcagtg aagcagctgc aagtaccgct gttgtgattg ctggccgttc gctaaacccc 1320
aacagggtga ctttcaaggc caacaggcct ttcctggttt ttataagaga agttcctctg 1380
aacactatta tcttcatggg cagagtagcc aacccttgtg ttaagggagg cggaagccat 1440
caccaccatc accactaa 1458
<210>614
<211>485
<212>PRT
<213>Artificial Sequence
<220>
<223>AT3 His tag fusion protein
<400>614
Met Leu Leu Val Asn Gln Ser His Gln Gly Phe Asn Lys Glu His Thr
1 5 10 15
Ser Lys Met Val Ser Ala Ile Val Leu Tyr Val Leu Leu Ala Ala Ala
20 25 30
Ala His Ser Ala Phe Ala Ala Asp Leu Gly Ser His Gly Ser Pro Val
35 40 45
Asp Ile Cys Thr Ala Lys Pro Arg Asp Ile Pro Met Asn Pro Met Cys
50 55 60
Ile Tyr Arg Ser Pro Glu Lys Lys Ala Thr Glu Asp Glu Gly Ser Glu
65 70 75 80
Gln Lys Ile Pro Glu Ala Thr Asn Arg Arg Val Trp Glu Leu Ser Lys
85 90 95
Ala Asn Ser Arg Phe Ala Thr Thr Phe Tyr Gln His Leu Ala Asp Ser
100 105 110
Lys Asn Asp Asn Asp Asn Ile Phe Leu Ser Pro Leu Ser Ile Ser Thr
115 120 125
Ala Phe Ala Met Thr Lys Leu Gly Ala Cys Asn Asp Thr Leu Gln Gln
130 135 140
Leu Met Glu Val Phe Lys Phe Asp Thr Ile Ser Glu Lys Thr Ser Asp
145 150 155 160
Gln Ile His Phe Phe Phe Ala Lys Leu Asn Cys Arg Leu Tyr Arg Lys
165 170 175
Ala Asn Lys Ser Ser Lys Leu Val Ser Ala Asn Arg Leu Phe Gly Asp
180 185 190
Lys Ser Leu Thr Phe Asn Glu Thr Tyr Gln Asp Ile Ser Glu Leu Val
195 200 205
Tyr Gly Ala Lys Leu Gln Pro Leu Asp Phe Lys Glu Asn Ala Glu Gln
210 215 220
Ser Arg Ala Ala Ile Asn Lys Trp Val Ser Asn Lys Thr Glu Gly Arg
225 230 235 240
Ile Thr Asp Val Ile Pro Ser Glu Ala Ile Asn Glu Leu Thr Val Leu
245 250 255
Val Leu Val Asn Thr Ile Tyr Phe Lys Gly Leu Trp Lys Ser Lys Phe
260 265 270
Ser Pro Glu Asn Thr Arg Lys Glu Leu Phe Tyr Lys Ala Asp Gly Glu
275 280 285
Ser Cys Ser Ala Ser Met Met Tyr Gln Glu Gly Lys Phe Arg Tyr Arg
290 295 300
Arg Val Ala Glu Gly Thr Gln Val Leu Glu Leu Pro Phe Lys Gly Asp
305 310 315 320
Asp Ile Thr Met Val Leu Ile Leu Pro Lys Pro Glu Lys Ser Leu Ala
325 330 335
Lys Val Glu Lys Glu Leu Thr Pro Glu Val Leu Gln Glu Trp Leu Asp
340 345 350
Glu Leu Glu Glu Met Met Leu Val Val His Met Pro Arg Phe Arg Ile
355 360 365
Glu Asp Gly Phe Ser Leu Lys Glu Gln Leu Gln Asp Met Gly Leu Val
370 375 380
Asp Leu Phe Ser Pro Glu Lys Ser Lys Leu Pro Gly Ile Val Ala Glu
385 390 395 400
Gly Arg Asp Asp Leu Tyr Val Ser Asp Ala Phe His Lys Ala Phe Leu
405 410 415
Glu Val Asn Glu Glu Gly Ser Glu Ala Ala Ala Ser Thr Ala Val Val
420 425 430
Ile Ala Gly Arg Ser Leu Asn Pro Asn Arg Val Thr Phe Lys Ala Asn
435 440 445
Arg Pro Phe Leu Val Phe Ile Arg Glu Val Pro Leu Asn Thr Ile Ile
450 455 460
Phe Met Gly Arg Val Ala Asn Pro Cys Val Lys Gly Gly Gly Ser His
465 470 475 480
His His His His His
485
<210>615
<211>6644
<212>DNA
<213>Artificial Sequence
<220>
<223>pMal2c vector
<400>615
ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga 60
gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg 120
gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa 180
cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac 240
aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc 300
acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg 360
tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc 420
ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca 480
ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga 540
cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc 600
tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg 660
cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag 720
cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga 780
atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa 840
tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg 900
acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc 960
gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga 1020
agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata 1080
cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 1140
cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag 1200
gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg 1260
tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg 1320
tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt 1380
ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga 1440
attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga 1500
gcacttcacc aacaaggacc atagattatg aaaatcgaag aaggtaaact ggtaatctgg 1560
attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat 1620
accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt 1680
gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac 1740
gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat 1800
ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt 1860
gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa 1920
gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac 1980
ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag 2040
tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg 2100
ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac 2160
tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg 2220
gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc 2280
aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt 2340
ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg 2400
gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag 2460
ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg 2520
ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2580
gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2640
aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcagaattc 2700
ggatcctcta gagtcgacct gcaggcaagc ttggcactgg ccgtcgtttt acaacgtcgt 2760
gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc 2820
agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg 2880
aatggcgaat ggcagcttgg ctgttttggc ggatgagata agattttcag cctgatacag 2940
attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg 3000
gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt 3060
gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca 3120
gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag 3180
gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc 3240
aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg 3300
cctttttgcg tttctacaaa ctctttttgt ttatttttct aaatacattc aaatatgtat 3360
ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 3420
agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 3480
tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 3540
gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 3600
gaacgttctc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 3660
gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 3720
gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 3780
agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 3840
ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 3900
cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 3960
gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 4020
cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 4080
gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 4140
ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 4200
acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 4260
ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 4320
ccccggttga taatcagaaa agccccaaaa acaggaagat tgtataagca aatatttaaa 4380
ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt 4440
ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag 4500
ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg 4560
tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat 4620
caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc 4680
gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga 4740
aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac 4800
ccgccgcgct taatgcgccg ctacagggcg cgtaaaagga tctaggtgaa gatccttttt 4860
gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 4920
gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 4980
caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 5040
ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg 5100
tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 5160
ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 5220
tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 5280
cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 5340
gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 5400
ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 5460
gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 5520
agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 5580
tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 5640
tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc 5700
gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca 5760
caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat 5820
acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg 5880
ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg 5940
tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc 6000
tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg 6060
cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca 6120
tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt 6180
tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg 6240
atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc 6300
gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg 6360
ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg 6420
ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg 6480
ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt 6540
cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca 6600
cgatcatgcg cacccgtggc caggacccaa cgctgcccga aatt 6644
<210>616
<211>543
<212>DNA
<213>M13phage
<220>
<223>GeneIII
<400>616
agcggctcga gcggtggtgg ctctgagggt ggcggttctg agggtggcgg ctctgaggga 60
ggcggttccg gtggtggctc tggttccggt gattttgatt atgaaaagat ggcaaacgct 120
aataaggggg ctatgaccga aaatgccgat gaaaacgcgc tacagtctga cgctaaaggc 180
aaacttgatt ctgtcgctac tgattacggt gctgctatcg atggtttcat tggtgacgtt 240
tccggccttg ctaatggtaa tggtgctact ggtgattttg ctggctctaa ttcccaaatg 300
gctcaagtcg gtgacggtga taattcacct ttaatgaata atttccgtca atatttacct 360
tccctccctc aatcggttga atgtcgccct tttgtctttg gcgctggtaa accgtatgaa 420
ttttctattg attgtgacaa aataaactta ttccgtggtg tctttgcgtt tcttttatat 480
gttgccacct ttatgtatgt attttctacg tttgctaaca tactgcgtaa taaggagtct 540
taa 543
<210>617
<211>3
<212>PRT
<213>Artificial Sequence
<220>
<223>uPA cleavage substrate AGR
<400>617
Ala Gly Arg
1
<210>618
<211>20
<212>DNA
<213>Artificial Sequence
<220>
<223>MTSP sequencing primer
<400>618
ggtgttttca cgagcacttc 20
<210>619
<211>6601
<212>DNA
<213>Artificial Sequence
<220>
<223>pCAT0002vector
<400>619
gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat 60
tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg tcgggattaa 120
acgacatttt cgaagcgcag aagattgaat ggcacgaggc gagcgaaaac ttgtactttc 180
agggcgtgca tcatccgccg agctatgtag cgcatctggc ttccgacttt ggcgtgcgtg 240
tgtttcagca ggtcgcccag gcgtcaaaag atcgcaatgt ggtgttcagc ccgtatggtg 300
ttgcgtccgt gttggcgatg cttcaactta ctaccggcgg tgaaacgcag caacagattc 360
aggcggcgat gggcttcaag atagatgaca aaggcatggc acccgccctg cgtcatctgt 420
ataaagaact gatggggccg tggaacaaag atgagattag caccaccgat gcgatatttg 480
tgcagcgcga tctgaagctg gtccagggat tcatgcctca tttcttccgt ctgtttcgta 540
gcaccgtgaa acaggtggat tttagcgagg tggagcgtgc ccgcttcatc ataaacgatt 600
gggtgaagac ccacacgaag ggcatgattt cgcacctgct tgggaccggc gctgtggatc 660
aactgacccg tctggtgctg gtaaatgcgt tgtatttcaa cgggcaatgg aaaaccccgt 720
ttccggattc aagcacccat cgtcgtcttt ttcataaaag cgatggctct accgtcagcg 780
tccccatgat ggcgcagacg aacaaattca attacaccga attcaccacg ccggatggcc 840
actactatga tattctggag ctgccgtacc atggcgatac cctgagcatg ttcattgcgg 900
caccgtatga gaaagaagtg ccgctgtcgg ctctgaccaa cattctgagc gcgcagctta 960
ttagccattg gaaaggcaat atgacccgcc tgccacgtct gctggttctg ccgaaattta 1020
gcctggagac cgaagtggat ctgcgtaaac cgctggagaa tctggggatg acggatatgt 1080
tccggcagtt tcaagctgat ttcaccagcc ttagcgacca ggaacctctg catgtggccc 1140
tggcgttgca gaaggttaag atcgaagtga atgaaagcgg aaccgtggcg tctagtagca 1200
ctgctgtgat tcgtcgtgcg cgtatggcgc ctgaggaaat tatcatcgac cgtccgttcc 1260
tgtttgtggt gcgtcacaat ccgaccggga ctgtgctgtt tatgggccag gtgatggaac 1320
cgtaataagc ttgcggccgc actcgagcac caccaccacc accactgaga tccggctgct 1380
aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa 1440
ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc 1500
ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 1560
cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 1620
cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 1680
tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 1740
gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 1800
cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 1860
attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga 1920
tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt tcaggtggca 1980
cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 2040
tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 2100
gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 2160
ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 2220
cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 2280
ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 2340
cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 2400
tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 2460
tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 2520
tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 2580
ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 2640
tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 2700
cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 2760
gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 2820
ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 2880
acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 2940
cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 3000
atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 3060
tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 3120
tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 3180
aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 3240
aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt 3300
taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 3360
taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 3420
agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 3480
tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 3540
cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 3600
agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 3660
gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 3720
aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 3780
tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag 3840
ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg 3900
aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat 3960
atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg 4020
ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg 4080
ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg 4140
agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa 4200
agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc 4260
tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg 4320
gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg 4380
gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat 4440
gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag 4500
agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag 4560
ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc 4620
cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc 4680
gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc 4740
taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg 4800
cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg 4860
gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac 4920
aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct 4980
gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata 5040
gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt 5100
cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc 5160
gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 5220
agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa 5280
cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt 5340
ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct 5400
gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc 5460
ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg 5520
aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc 5580
gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc 5640
cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg 5700
acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat 5760
actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc 5820
agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac 5880
gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac 5940
catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat 6000
ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt 6060
gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc 6120
cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt 6180
ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac 6240
caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca 6300
ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc 6360
ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga 6420
tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc 6480
tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc 6540
cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg 6600
a 6601
<210>620
<211>4
<212>PRT
<213>Artificial Sequence
<220>
<223>GGGS linker
<400>620
Gly Gly Gly Ser
1
<210>621
<211>405
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant PAI-1
<223>OptiPAI-1stab
<400>621
Met Ser Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp His
1 5 10 15
Glu Ala Ser Glu Asn Leu Tyr Phe Gln Gly Val His His Pro Pro Ser
20 25 30
Tyr Val Ala His Leu Ala Ser Asp Phe Gly Val Arg Val Phe Gln Gln
35 40 45
Val Ala Gln Ala Ser Lys Asp Arg Asn Val Val Phe Ser Pro Tyr Gly
50 55 60
Val Ala Ser Val Leu Ala Met Leu Gln Leu Thr Thr Gly Gly Glu Thr
65 70 75 80
Gln Gln Gln Ile Gln Ala Ala Met Gly Phe Lys Ile Asp Asp Lys Gly
85 90 95
Met Ala Pro Ala Leu Arg His Leu Tyr Lys Glu Leu Met Gly Pro Trp
100 105 110
Asn Lys Asp Glu Ile Ser Thr Thr Asp Ala Ile Phe Val Gln Arg Asp
115 120 125
Leu Lys Leu Val Gln Gly Phe Met Pro His Phe Phe Arg Leu Phe Arg
130 135 140
Ser Thr Val Lys Gln Val Asp Phe Ser Glu Val Glu Arg Ala Arg Phe
145 150 155 160
Ile Ile Asn Asp Trp Val Lys Thr His Thr Lys Gly Met Ile Ser His
165 170 175
Leu Leu Gly Thr Gly Ala Val Asp Gln Leu Thr Arg Leu Val Leu Val
180 185 190
Asn Ala Leu Tyr Phe Asn Gly Gln Trp Lys Thr Pro Phe Pro Asp Ser
195 200 205
Ser Thr His Arg Arg Leu Phe His Lys Ser Asp Gly Ser Thr Val Ser
210 215 220
Val Pro Met Met Ala Gln Thr Asn Lys Phe Asn Tyr Thr Glu Phe Thr
225 230 235 240
Thr Pro Asp Gly His Tyr Tyr Asp Ile Leu Glu Leu Pro Tyr His Gly
245 250 255
Asp Thr Leu Ser Met Phe Ile Ala Ala Pro Tyr Glu Lys Glu Val Pro
260 265 270
Leu Ser Ala Leu Thr Asn Ile Leu Ser Ala Gln Leu Ile Ser His Trp
275 280 285
Lys Gly Asn Met Thr Arg Leu Pro Arg Leu Leu Val Leu Pro Lys Phe
290 295 300
Ser Leu Glu Thr Glu Val Asp Leu Arg Lys Pro Leu Glu Asn Leu Gly
305 310 315 320
Met Thr Asp Met Phe Arg Gln Phe Gln Ala Asp Phe Thr Ser Leu Ser
325 330 335
Asp Gln Glu Pro Leu His Val Ala Leu Ala Leu Gln Lys Val Lys Ile
340 345 350
Glu Val Asn Glu Ser Gly Thr Val Ala Ser Ser Ser Thr Ala Val Ile
355 360 365
Arg Arg Ala Arg Met Ala Pro Glu Glu Ile Ile Ile Asp Arg Pro Phe
370 375 380
Leu Phe Val Val Arg His Asn Pro Thr Gly Thr Val Leu Phe Met Gly
385 390 395 400
Gln Val Met Glu Pro
405
<210>622
<211>381
<212>PRT
<213>Homo sapiens
<220>
<223>Mutant PAI-1
<223>V1C OptiPAI-lstab
<400>622
Met Cys Val His His Pro Pro Ser Tyr Val Ala His Leu Ala Ser Asp
1 5 10 15
Phe Gly Val Arg Val Phe Gln Gln Val Ala Gln Ala Ser Lys Asp Arg
20 25 30
Asn Val Val Phe Ser Pro Tyr Gly Val Ala Ser Val Leu Ala Met Leu
35 40 45
Gln Leu Thr Thr Gly Gly Glu Thr Gln Gln Gln Ile Gln Ala Ala Met
50 55 60
Gly Phe Lys Ile Asp Asp Lys Gly Met Ala Pro Ala Leu Arg His Leu
65 70 75 80
Tyr Lys Glu Leu Met Gly Pro Trp Asn Lys Asp Glu Ile Ser Thr Thr
85 90 95
Asp Ala Ile Phe Val Gln Arg Asp Leu Lys Leu Val Gln Gly Phe Met
100 105 110
Pro His Phe Phe Arg Leu Phe Arg Ser Thr Val Lys Gln Val Asp Phe
115 120 125
Ser Glu Val Glu Arg Ala Arg Phe Ile Ile Asn Asp Trp Val Lys Thr
130 135 140
His Thr Lys Gly Met Ile Ser His Leu Leu Gly Thr Gly Ala Val Asp
145 150 155 160
Gln Leu Thr Arg Leu Val Leu Val Asn Ala Leu Tyr Phe Ash Gly Gln
165 170 175
Trp Lys Thr Pro Phe Pro Asp Ser Ser Thr His Arg Arg Leu Phe His
180 185 190
Lys Ser Asp Gly Ser Thr Val Ser Val Pro Met Met Ala Gln Thr Asn
195 200 205
Lys Phe Ash Tyr Thr Glu Phe Thr Thr Pro Asp Gly His Tyr Tyr Asp
210 215 220
Ile Leu Glu Leu Pro Tyr His Gly Asp Thr Leu Ser Met Phe Ile Ala
225 230 235 240
Ala Pro Tyr Glu Lys Glu Val Pro Leu Ser Ala Leu Thr Asn Ile Leu
245 250 255
Ser Ala Gln Leu Ile Ser His Trp Lys Gly Asn Met Thr Arg Leu Pro
260 265 270
Arg Leu Leu Val Leu Pro Lys Phe Ser Leu Glu Thr Glu Val Asp Leu
275 280 285
Arg Lys Pro Leu Glu Asn Leu Gly Met Thr Asp Met Phe Arg Gln Phe
290 295 300
Gln Ala Asp Phe Thr Ser Leu Ser Asp Gln Glu Pro Leu His Val Ala
305 310 315 320
Leu Ala Leu Gln Lys Val Lys Ile Glu Val Asn Glu Ser Gly Thr Val
325 330 335
Ala Ser Ser Ser Thr Ala Val Ile Arg Arg Ala Arg Met Ala Pro Glu
340 345 350
Glu Ile Ile Ile Asp Arg Pro Phe Leu Phe Val Val Arg His Asn Pro
355 360 365
Thr Gly Thr Val Leu Phe Met Gly Gln Val Met Glu Pro
370 375 380
<210>623
<211>6526
<212>DNA
<213>Artificial Sequence
<220>
<223>pCAT0051 vector
<400>623
gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat 60
tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg tgccatcatc 120
cgccgagcta tgtagcgcat ctggcttccg actttggcgt gcgtgtgttt cagcaggtcg 180
cccaggcgtc aaaagatcgc aatgtggtgt tcagcccgta tggtgttgcg tccgtgttgg 240
cgatgcttca acttactacc ggcggtgaaa cgcagcaaca gattcaggcg gcgatgggct 300
tcaagataga tgacaaaggc atggcacccg ccctgcgtca tctgtataaa gaactgatgg 360
ggccgtggaa caaagatgag attagcacca ccgatgcgat atttgtgcag cgcgatctga 420
agctggtcca gggattcatg cctcatttct tccgtctgtt tcgtagcacc gtgaaacagg 480
tggattttag cgaggtggag cgtgcccgct tcatcataaa cgattgggtg aagacccaca 540
cgaagggcat gatttcgcac ctgcttggga ccggcgctgt ggatcaactg acccgtctgg 600
tgctggtaaa tgcgttgtat ttcaacgggc aatggaaaac cccgtttccg gattcaagca 660
cccatcgtcg tctttttcat aaaagcgatg gctctaccgt cagcgtcccc atgatggcgc 720
agacgaacaa attcaattac accgaattca ccacgccgga tggccactac tatgatattc 780
tggagctgcc gtaccatggc gataccctga gcatgttcat tgcggcaccg tatgagaaag 840
aagtgccgct gtcggctctg accaacattc tgagcgcgca gcttattagc cattggaaag 900
gcaatatgac ccgcctgcca cgtctgctgg ttctgccgaa atttagcctg gagaccgaag 960
tggatctgcg taaaccgctg gagaatctgg ggatgacgga tatgttccgg cagtttcaag 1020
ctgatttcac cagccttagc gaccaggaac ctctgcatgt ggccctggcg ttgcagaagg 1080
ttaagatcga agtgaatgaa agcggaaccg tggcgtctag tagcactgct gtgattcgtc 1140
gtgcgcgtat ggcgcctgag gaaattatca tcgaccgtcc gttcctgttt gtggtgcgtc 1200
acaatccgac cgggactgtg ctgtttatgg gccaggtgat ggaaccgtaa taagcttgcg 1260
gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag 1320
gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct 1380
aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg 1440
acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 1500
ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 1560
cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 1620
gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 1680
catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 1740
gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 1800
aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 1860
acgcgaattt taacaaaata ttaacgttta caatttcagg tggcactttt cggggaaatg 1920
tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 1980
gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac 2040
atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 2100
cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca 2160
tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc 2220
caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 2280
ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac 2340
cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 2400
taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg 2460
agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 2520
cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg 2580
caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat 2640
taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 2700
ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 2760
cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 2820
aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 2880
attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt 2940
tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 3000
aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 3060
gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 3120
cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 3180
gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca 3240
agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 3300
ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 3360
cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 3420
acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 3480
gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 3540
ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 3600
agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 3660
cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 3720
tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc 3780
gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc 3840
ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag 3900
tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac 3960
tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 4020
ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 4080
aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg 4140
tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc 4200
agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt 4260
ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg 4320
aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa 4380
cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag 4440
ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat 4500
cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt 4560
tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag 4620
cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa 4680
ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc 4740
gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg 4800
aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc 4860
gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct 4920
acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc 4980
caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc 5040
taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 5100
aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 5160
attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt 5220
caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg 5280
aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc 5340
gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat 5400
tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt 5460
cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc 5520
tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc 5580
cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag 5640
atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt 5700
ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat 5760
ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag 5820
attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac 5880
gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg 5940
cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg 6000
tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt 6060
tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc 6120
ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact 6180
ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg 6240
gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga 6300
ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc 6360
ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt 6420
ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg 6480
tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcga 6526
<210>624
<211>4202
<212>DNA
<213>Artificial Sequence
<220>
<223>pQE30 vector
<400>624
ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca 60
attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga 120
ggatcgcatc accatcacca tcacggatcc gatgagaagg actgcgactg tgggctgcgg 180
tcattcacga gacaggctcg tgttgttggg ggcacggatg cggatgaggg cgagtggccc 240
tggcaggtaa gcctgcatgc tctgggccag ggccacatct gcggtgcttc cctcatctct 300
cccaactggc tggtctctgc cgcacactgc tacatcgatg acagaggatt caggtactca 360
gaccccacgc agtggacggc cttcctgggc ttgcacgacc agagccagcg cagcgcccct 420
ggggtgcagg agcgcaggct caagcgcatc atctcccacc ccttcttcaa tgacttcacc 480
ttcgactatg acatcgcgct gctggagctg gagaaaccgg cagagtacag ctccatggtg 540
cggcccatct gcctgccgga cgcctcccat gtcttccctg ccggcaaggc catctgggtc 600
acgggctggg gacacaccca gtatggaggc actggcgcgc tgatcctgca aaagggtgag 660
atccgcgtca tcaaccagac cacctgcgag aacctcctgc cgcagcagat cacgccgcgc 720
atgatgtgcg tgggcttcct cagcggcggc gtggactcct gccagggtga ttccggggga 780
cccctgtcca gcgtggaggc ggatgggcgg atcttccagg ccggtgtggt gagctgggga 840
gacggctgcg ctcagaggaa caagccaggc gtgtacacaa ggctccctct gtttcgggac 900
tggatcaaag agaacactgg ggtatagaag cttaattagc tgagcttgga ctcctgttga 960
tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc 1020
gggcgttttt tattggtgag aatccaagct agcttggcga gattttcagg agctaaggaa 1080
gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt 1140
aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag 1200
ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc 1260
tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa 1320
gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa 1380
actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac 1440
atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt 1500
attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta 1560
aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg 1620
caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc 1680
ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg 1740
gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggggtaat gactctctag 1800
cttgaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg 1860
ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccctctagag ctgcctcgcg 1920
cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct 1980
tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc 2040
gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta 2100
actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc 2160
acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact 2220
cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 2280
ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 2340
aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 2400
acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 2460
gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 2520
ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 2580
gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 2640
cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 2700
taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 2760
atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga 2820
cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 2880
cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 2940
ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 3000
ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 3060
tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 3120
aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 3180
tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 3240
gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 3300
atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 3360
tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 3420
ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 3480
ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 3540
tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 3600
ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 3660
ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 3720
tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 3780
gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 3840
taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 3900
cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 3960
agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 4020
gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 4080
ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa 4140
ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc 4200
ac 4202
<210>625
<211>4822
<212>DNA
<213>Artificial Sequence
<220>
<223>pPAI-his vector
<220>
<221>misc_feature
<222>(1)...(4822)
<223>n ＝A，T，C or G
<400>625
gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct 60
agaaataatt ttgtttaact ttaagaagga gatatacata tgcaccatca ccatcaccat 120
gctagcgaga atctttattt tcagggcgtg caccatcccc catcctacgt ggcccacctg 180
gcctcagact tcggggtgag ggtgtttcag caggtggcgc aggcctccaa ggaccgcaac 240
gtggttttct caccctatgg ggtggcctcg gtgttggcca tgctccagct gacaacagga 300
ggagaaaccc agcagcagat tcaagcagct atgggattca agattgatga caagggcatg 360
gcccccgccc tccggcatct gtacaaggag ctcatggggc catggaacaa ggatgagatc 420
agcaccacag acgcgatctt cgtccagcgg gatctgaagc tggtccaggg cttcatgccc 480
cacttcttca ggctgttccg gagcacggtc aagcaagtgg acttttcaga ggtggagaga 540
gccagattca tcatcaatga ctgggtgaag acacacacaa aaggtatgat cagcaacttg 600
cttgggaaag gagccgtgga ccagctgaca cggctggtgc tggtgaatgc cctctacttc 660
aacggccagt ggaagactcc cttccccgac tccagcaccc accgccgcct cttccacaaa 720
tcagacggca gcactgtctc tgtgcccatg atggctcaga ccaacaagtt caactatact 780
gagttcacca cgcccgatgg ccattactac gacatcctgg aactgcccta ccacggggac 840
accctcagca tgttcattgc tgccccttat gaaaaagagg tgcctctctc tgccctcacc 900
aacattctga gtgcccagct catcagccac tggaaaggca acatgaccag gctgccccgc 960
ctcctggttc tgcccaagtt ctccctggag actgaagtcg acctcaggaa gcccctagag 1020
aacctgggaa tgaccgacat gttcagacag tttcaggctg acttcacgag tctttcagac 1080
caagagcctc tccacgtcgc gcaggcgctg cagaaagtga agatcgaggt gaacgagagt 1140
ggcacggtgg cctcctcatc cacagctgtc atannnnnnn nnnnnnnnnn ncccgaggag 1200
atcatcatgg acagaccctt cctctttgtg gtccggcaca accccacagg aacagtcctt 1260
ttcatgggcc aagtgatgga accctgaaag cttccgggat ccggctgcta acaaagcccg 1320
aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc 1380
ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gatcgacgcg 1440
aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc 1500
gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc 1560
gctcgcggct cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta 1620
tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct 1680
tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga 1740
agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg 1800
cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc 1860
tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg ggtgcgcatg 1920
atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag 1980
aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc 2040
tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag 2100
tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt 2160
ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg 2220
tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt 2280
tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc 2340
atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt 2400
aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg 2460
gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac 2520
cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 2580
gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 2640
tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga 2700
gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc 2760
ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt 2820
cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 2880
caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 2940
caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 3000
ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 3060
cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 3120
ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 3180
tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 3240
gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 3300
ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 3360
ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 3420
gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 3480
aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 3540
tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 3600
ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 3660
tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 3720
aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 3780
tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 3840
ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 3900
gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 3960
gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 4020
taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg 4080
tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 4140
ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 4200
tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 4260
ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 4320
tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata 4380
ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 4440
aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 4500
actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 4560
aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 4620
tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 4680
aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 4740
ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga 4800
ggccctttcg tcttcaagaa tt 4822
<210>626
<211>42
<212>DNA
<213>Artificial Sequence
<220>
<223>ATIII cDNA forward primer
<400>626
gtcactgact gacgtggatc ccacgggagc cctgtggaca tc 42
<210>627
<211>9
<212>PRT
<213>Artificial Sequence
<220>
<223>ATIII cDNA forward primer -encoded protein
<400>627
Gly Ser His Gly Ser Pro Val Asp Ile
1 5
<210>628
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>ATIII cDNA reverse primer
<400>628
gtagccaacc cttgtgttaa gggaggcgga agccatcacc accatcacca ctaagaattc 60
<210>629
<211>17
<212>PRT
<213>Artificial Sequence
<220>
<223>ATIII cDNA reverse primer-encoded protein
<400>629
Val Ala Asn Pro Cys Val Lys Gly Gly Gly Ser His His His His His
1 5 10 15
His
<210>630
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Wild type AT3primer
<400>630
gctgcaagta ccgctgttgt gattgctggc cgttcgctaa accccaacag ggtgactttc 60
<210>631
<211>20
<212>PRT
<213>Artificial Sequence
<220>
<223>Wild type AT3 primer-encoded protein
<400>631
Ala Ala Ser Thr Ala Val Val Ile Ala Gly Arg Ser Leu Asn Pro Asn
1 5 10 15
Arg Val Thr Phe
20
<210>632
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Complement C2 Target Sequence primer
<400>632
gctgcaagta ccgctgttgt gtcgttaggc cgtaaaatta accccaacag ggtgactttc 60
<210>633
<211>20
<212>PRT
<213>Artificial Sequence
<220>
<223>Complement C2 Target Sequence primer-encoded
protein
<400>633
Ala Ala Ser Thr Ala Val Val Ser Leu Gly Arg Lys Ile Asn Pro Asn
1 5 10 15
Arg Val Thr Phe
20
<210>634
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>VEGFR2 Target Sequence primer
<400>634
gctgcaagta ccgctgttgt gcgccgtgtg cgcaaagaaa accccaacag ggtgactttc 60
<210>635
<211>20
<212>PRT
<213>Artificial Sequence
<220>
<223>VEGFR2 Target Sequence primer-encoded primer
<400>635
Ala Ala Ser Thr Ala Val Val Arg Arg Val Arg Lys Glu Asn Pro Asn
1 5 10 15
Arg Val Thr Phe
20
<210>636
<211>68
<212>DNA
<213>Escherichia coli
<220>
<223>STII leader
<400>636
tgaaaaagaa tatcgcattt cttcttgcat ctatgttcgt tttttctatt gctacaaacg 60
cgtatgca 68
Claims (170)
1.一种方法，其包括：
a)将蛋白酶和/或蛋白酶的催化活性部分的集合与蛋白酶捕获多肽接触，在所述蛋白酶捕获多肽由所述蛋白酶或其催化活性部分裂解时，形成含有所述蛋白酶捕获多肽与所述集合中蛋白酶或者其催化活性部分的稳定复合物；及
b)鉴别或选择复合物中的蛋白酶或其催化活性部分。
2.权利要求1的方法，其中所述蛋白酶捕获多肽被修饰为由对于靶底物具有预定底物特异性和/或活性的蛋白酶裂解。
3.权利要求2的方法，其中：
所述鉴别或选择的蛋白酶与模板蛋白酶相比是被修饰的；
所述鉴别或选择的蛋白酶与不含有所述修饰的模板蛋白酶相比对于靶底物具有改变的活性和/或特异性。
4.权利要求1-3任一项的方法，进一步包括从所述集合的未复合成员中分离出复合的蛋白酶。
5.权利要求1-3任一项的方法，其中所述复合物中的蛋白酶捕获多肽与蛋白酶是共价连接的。
6.权利要求1-5任一项的方法，其中：
所述蛋白酶捕获多肽被标记以进行检测或分离；及
所述分离通过捕获含有可检测的蛋白酶捕获多肽与任何蛋白酶或其催化活性部分的复合物而实现。
7.权利要求6的方法，其中所述捕获是在悬浮液、溶液或者在固体支持物上实现的。
8.权利要求7的方法，其中：
使所述蛋白酶捕获多肽附着于固体支持物；及
在所述蛋白酶捕获多肽与蛋白酶或其催化活性部分的集合接触之前、期间或之后实现附着。
9.权利要求8的方法，其中所述固体支持物是酶联免疫吸附测定(ELISA)平板的孔。
10.权利要求6的方法，其中所述蛋白酶捕获多肽用生物素标记。
11.权利要求6的方法，其中：
所述蛋白酶捕获剂用His标记进行标记；及
所述分离通过用金属螯合剂捕获而实现的。
12.权利要求11的方法，其中所述金属螯合剂选自硫酸镍(NiSO4)、氯化钴(CoCl2)、硫酸铜(CuSO4)和氯化锌(ZnCl2)。
13.权利要求1-12任一项的方法，进一步包括扩增分离的复合物中所述蛋白酶或其催化活性部分。
14.权利要求13的方法，其中分离的复合物中所述蛋白酶或其催化活性部分在噬菌体上被展示，扩增通过用噬菌体感染宿主细胞实现。
15.权利要求14的方法，其中所述宿主细胞是细菌。
16.权利要求14的方法，进一步包括筛选扩增的蛋白酶对于靶底物的特异性和/或活性，其中特异性和/或活性在细菌细胞培养基、细菌周质、噬菌体上清或者在纯化的蛋白质上评定。
17.权利要求16的方法，其中所述靶底物是疾病或病症病因学中涉及的多肽或多肽中的裂解序列。
18.权利要求1-17任一项的方法，其是多元的，其中：
将多个不同的蛋白酶捕获多肽与所述集合接触；及
至少两种蛋白酶捕获多肽可以被可鉴别地检测，从而
鉴别一个以上的蛋白酶。
19.权利要求1-18任一项的方法，其中所述复合物中蛋白酶或其催化活性部分被展示，从而在鉴别或选择之后其可被扩增以产生蛋白酶和/或其催化活性部分的第二个集合。
20.权利要求19的方法，进一步包括将所述第二个集合与第二种蛋白酶捕获多肽接触以产生第二组稳定复合物。
21.权利要求20的方法，其中所述第二种蛋白酶捕获多肽与第一种蛋白酶捕获多肽相同或不同。
22.权利要求20的方法，进一步包括鉴别或选择所述第二组稳定复合物中的蛋白酶。
23.权利要求1-22任一项的方法，其中所述蛋白酶捕获多肽选自丝氨酸蛋白酶抑制蛋白、α巨球蛋白家族成员或者p35家族成员。
24.权利要求4和6-23任一项的方法，其中所述稳定复合物包含与蛋白酶或其催化活性部分共价连接的蛋白酶捕获多肽。
25.一种筛选蛋白酶以鉴别具有预定底物特异性的任何蛋白酶的方法，包括：
a)将蛋白酶和/或蛋白酶催化活性部分的集合与蛋白酶捕获多肽接触，在蛋白酶捕获多肽裂解时形成所述蛋白酶捕获多肽与蛋白酶的稳定复合物，其中所述蛋白酶捕获多肽被选择或修饰为由具有预定底物特异性的蛋白酶裂解；及
b)鉴别复合物中蛋白酶或其催化活性部分，从而选择具有预定底物特异性的蛋白酶。
26.权利要求1-25任一项的方法，其中所述集合包含至少5、10、50、100、500、103、104、105、106或更多个不同成员。
27.权利要求1-26任一项的方法，其中所述蛋白酶或其催化活性部分是丝氨酸和/或半胱氨酸蛋白酶。
28.权利要求1-27任一项的方法，其中所述蛋白酶或其催化活性部分被展示。
29.权利要求28的方法，其中所述蛋白酶或其催化活性部分在固体支持物、细胞表面或者在微生物表面上被展示。
30.权利要求29的方法，其中所述蛋白酶或其催化活性部分在酵母、细菌、病毒、噬菌体、核酸、mRNA分子或者在核糖体表面上被展示。
31.权利要求29的方法，其中所述微生物是大肠杆菌(E.coli)、酿酒酵母(S.cerevisiae)，或者选自M13、fd、T7或者杆状病毒的病毒。
32.权利要求1-31任一项的方法，其是均相反应。
33.权利要求1-28任一项的方法，其中所述蛋白酶或其催化活性部分在噬菌体展示文库中被展示。
34.权利要求1-33任一项的方法，其中所述蛋白酶以蛋白酶的催化活性部分提供。
35.权利要求1-34任一项的方法，其中：
将至少两种不同的蛋白酶捕获多肽与所述集合接触；及
至少一种蛋白酶捕获多肽是可检测地标记的。
36.权利要求35的方法，其中至少一种蛋白酶捕获多肽是可检测的，以实现捕获含有可检测的蛋白酶捕获多肽与蛋白酶或其催化活性部分的稳定复合物。
37.权利要求35的方法，其中：
将至少两种不同的蛋白酶捕获多肽与所述集合接触；
只有一种蛋白酶捕获多肽是可检测地标记的；及
另一种蛋白酶或者其它蛋白酶与所述可检测地标记的蛋白酶捕获多肽相比均过量存在。
38.权利要求35的方法，其中所述可检测的标记是荧光标记或His标记。
39.权利要求35的方法，其中所述可检测的标记是生物素。
40.权利要求1-38任一项的方法，其中所述蛋白酶或其催化活性部分选自蛋白酶的胰凝乳蛋白酶、枯草杆菌蛋白酶、胱天蛋白酶和木瓜蛋白酶家族。
41.权利要求1-40任一项的方法，其中所述蛋白酶是丝氨酸蛋白酶、半胱氨酸蛋白酶或者其催化活性部分并选自粒酶B、testisin、trypstase β1、激肽释放酶hk5、corin、激肽释放酶12、DESC1 oritesase、trypstase γ1、激肽释放酶hK14、乙酰透明质酸结合性丝氨酸蛋白酶、类胰蛋白酶、激肽释放酶hK15、胰蛋白酶、嗜中性粒细胞弹性蛋白酶、甘露聚糖结合性凝集素相关丝氨酸蛋白酶-3、组织蛋白酶G、成髓细胞蛋白酶、粒酶A、粒酶M、类胰凝乳蛋白酶、粒酶K、粒酶H、胰凝乳蛋白酶B、胰腺弹性蛋白酶、胰腺内肽酶E、胰腺弹性蛋白酶II、肠肽酶、胰凝乳蛋白酶C、prostasin、激肽释放酶1、激肽释放酶hK2、激肽释放酶3、mesotrypsin、因子XII、血浆激肽释放酶KLK3、因子XI、因子IX、因子VII、因子Xa、凝血酶、蛋白C、项体蛋白、hepsin、肝细胞生长因子激活物、尿纤溶酶原激活物(uPA)、组织纤溶酶原激活物(tPA)、纤溶酶、neurosin、神经胰蛋白酶、neuropsin、激肽释放酶hK10、epitheliasin、prostase、chymopasin、激肽释放酶11、MT-SP1、spinesin、组织蛋白酶L、组织蛋白酶V、组织蛋白酶K、组织蛋白酶S、组织蛋白酶F、组织蛋白酶B、木瓜蛋白酶、cruzain、枯草杆菌蛋白酶、thermitase、C5a肽酶、fervidolysin、lactocepin、弗林蛋白酶、kexin、胱天蛋白酶-1、胱天蛋白酶-3、胱天蛋白酶-7、胱天蛋白酶-6、胱天蛋白酶-2、胱天蛋白酶-4、胱天蛋白酶-5、胱天蛋白酶-8、胱天蛋白酶-9、胱天蛋白酶-10、胱天蛋白酶-11、胱天蛋白酶-12、胱天蛋白酶-1、胱天蛋白酶-13和胱天蛋白酶-14。
42.权利要求1-41任一项的方法，其中所述蛋白酶或其催化活性部分选自尿激酶纤溶酶原激活物(u-PA)、组织纤溶酶原激活物(t-PA)和MT-SP1。
43.权利要求1-42任一项的方法，其中所述蛋白酶捕获多肽选自丝氨酸蛋白酶抑制蛋白、p35家族成员、α-巨球蛋白家族成员或者其修饰形式。
44.权利要求1-42任一项的方法，其中所述蛋白酶捕获多肽是选自纤溶酶原激活物抑制剂-1(PAI-1)或者抗凝血酶III(AT3)的丝氨酸蛋白酶抑制蛋白或其修饰形式。
45.权利要求43的方法，其中所述蛋白酶捕获多肽是α2-巨球蛋白或者其修饰形式。
46.权利要求43-45任一项的方法，其中所述修饰形式在所述蛋白酶捕获多肽的反应位点含有一或多个氨基酸置换、缺失或取代。
47.权利要求46的方法，其中所述修饰是相应于靶底物的裂解序列的任一或多个氨基酸置换。
48.权利要求47的方法，其中所述靶底物参与疾病或病症的病因学。
49.权利要求47的方法，其中所述靶底物是VEGFR、补体蛋白或者组织纤溶酶原激活物(tPA)的底物。
50.权利要求49的方法，其中所述VEGFR是VEGFR2。
51.权利要求49的方法，其中所述补体蛋白是C2。
52.权利要求47的方法，其中相应于裂解序列的所述一或多个氨基酸修饰具有SEQ ID NO：389、479和498所示任一氨基酸序列。
53.权利要求46的方法，其中所述蛋白酶捕获多肽是丝氨酸蛋白酶抑制蛋白且所述一或多个氨基酸置换是在丝氨酸蛋白酶抑制蛋白多肽的反应位点环中。
54.权利要求53的方法，其中所述一或多个氨基酸置换是在P4-P2’任一或多个位置中。
55.权利要求54的方法，其中所述丝氨酸蛋白酶抑制蛋白具有SEQ IDNO：497、499、610和611所示任一氨基酸序列。
56.权利要求46的方法，其中所述蛋白酶捕获多肽是α2巨球蛋白且所述一或多个氨基酸置换是在所述多肽的诱饵区域中。
57.权利要求48的方法，其中所述鉴别或选择的蛋白酶或其催化活性部分与非靶底物相比对于靶底物具有改变的底物特异性。
58.权利要求57的方法，其中所述非靶底物是相应模板蛋白酶的底物。
59.权利要求57的方法，其中所述底物特异性增加1.5倍、2倍、5倍、10倍、50倍、100倍、200倍、300倍、400倍、500倍或更多倍。
60.权利要求1-59任一项的方法，其中多次进行鉴别蛋白酶的方法，从而所述过程是重复的。
61.权利要求60的方法，其中进行述方法直至所述蛋白酶对于靶底物中裂解序列与相应模板蛋白酶的非靶底物相比呈现增加的底物特异性和/或活性。
62.权利要求60的方法，其中至少在所述方法的第一次重复中鉴别多个不同的蛋白酶。
63.权利要求60-62任一项的方法，进一步包括对比鉴别的蛋白酶的氨基酸序列以鉴别热点，其中：
热点是修饰的基因座，其在鉴别的蛋白酶中的至少2、3、4、5或更多种蛋白酶中出现；及
如果所述集合含有修饰的蛋白酶或其催化活性部分，修饰的基因座是被修饰的基因座。
64.权利要求1-63任一项的方法，进一步包括：
在鉴别第一种蛋白酶或多种蛋白酶之后，制备第二个蛋白酶集合，其中鉴别的第一种蛋白酶用作模板以在蛋白酶序列或其催化活性部分中产生进一步突变，由此第二个集合的成员含有具有鉴别的第一种蛋白酶的突变和额外突变的多肽；
然后将第二个集合与第二种蛋白酶捕获多肽接触，所述第二种蛋白酶捕获多肽与用于分离第一种蛋白酶或多种蛋白酶的第一种蛋白酶捕获多肽相同或不同，其中所述蛋白酶捕获多肽被修饰为由具有希望的靶底物特异性的蛋白酶裂解；及
从复合物集合中鉴别第二种蛋白酶或蛋白酶的催化活性部分，从而第二种鉴别的蛋白酶与第一种鉴别的蛋白酶相比对于靶底物具有较高的活性或特异性。
65.权利要求64的方法，其中所述第二个蛋白酶集合与第一种鉴别的模板蛋白酶或蛋白酶催化活性部分的氨基酸序列相比含有随机或聚焦突变。
66.权利要求65的方法，其中所述突变是在热点位置的聚焦突变。
67.权利要求66的方法，其中所述热点位置是在丝氨酸蛋白酶中并选自基于胰凝乳蛋白酶编号的第30、73、89和155位。
68.权利要求1-67任一项的方法，其中所述稳定复合物的形成是通过控制一或多个参数而调节的，所述参数选自反应时间、温度、pH、离子强度、文库浓度和蛋白酶捕获多肽浓度。
69.权利要求1-68任一项的方法，其中所述方法进一步包括向蛋白酶捕获多肽与蛋白酶或其催化活性部分之间的反应中加入竞争剂，从而增强鉴别的蛋白酶或催化活性部分的选择性。
70.权利要求69的方法，其中所述竞争剂是人血清或血浆。
71.权利要求69的方法，其中所述竞争剂是细胞或组织提取物或者生物学液体如尿液或血液。
72.权利要求69的方法，其中所述竞争剂是纯化或部分纯化的野生型形式的蛋白酶捕获剂。
73.权利要求69的方法，其中所述竞争剂是纯化或部分纯化的野生型形式的蛋白酶捕获多肽或者蛋白酶捕获多肽的一或多种特异变体。
74.权利要求69-70任一项的方法，其中所述竞争剂是人血浆。
75.权利要求69-72任一项的方法，其中所述竞争剂是纯化的野生型形式蛋白酶捕获多肽或者是修饰的蛋白酶捕获多肽。
76.一种进化或选择或鉴别对于至少两种不同的裂解序列具有特异性/选择性和/或活性的蛋白酶或其催化活性部分的重复方法，包括：
a)将蛋白酶和/或蛋白酶催化活性部分的集合与第一种蛋白酶捕获多肽接触，在所述蛋白酶或其催化活性部分裂解所述蛋白酶捕获多肽时，形成含有第一种蛋白酶捕获多肽与所述集合中的蛋白酶或其催化活性部分的稳定复合物，其中接触是在存在竞争剂的条件下进行；
b)鉴别或选择与第一种蛋白酶捕获多肽形成复合物的蛋白酶或其催化活性部分；
c)将与第一种蛋白酶捕获多肽形成复合物的蛋白酶或其催化活性部分与第二种蛋白酶捕获多肽在存在竞争剂的条件下接触；及
d)鉴别或选择与第二种蛋白酶捕获多肽形成复合物的蛋白酶或其催化活性部分，
从而鉴别的、进化的或者选择的蛋白酶或其催化活性部分对于至少两个裂解序列具有底物特异性和/或裂解活性。
77.权利要求76的方法，其中所述至少两个裂解序列在一个靶底物中。
78.权利要求76的方法，其中所述至少两个裂解序列在两个不同的靶底物中。
79.权利要求76-78任一项的方法，其中所述第一种和第二种蛋白酶捕获多肽是不同的且每种均被修饰为由对于不同靶底物具有希望的底物特异性的蛋白酶裂解。
80.权利要求76-79任一项的方法，进一步包括重复至少一次步骤a)和b)或者a)-d)，直至分离对于至少两个裂解序列具有希望的或预定底物特异性和裂解活性的鉴别或选择的蛋白酶。
81.权利要求76的方法，其中所述蛋白酶集合在相应于模板蛋白酶的位置包含氨基酸突变。
82.权利要求81的方法，其中所述底物特异性和/或裂解活性与相应模板蛋白酶相比是增加的。
83.权利要求80的方法，其中对于至少一个裂解序列的底物特异性与相应模板蛋白酶相比是增加的。
84.权利要求76的方法，其中所述竞争剂选自人血清、人血浆、细胞或组织提取物、生物学液体如尿液或血液、纯化或部分纯化的野生型蛋白酶捕获剂及蛋白酶捕获多肽的一或多种特异变体。
85.一种蛋白酶选择方法，包括：
a)将蛋白酶或其催化活性部分集合与第一种蛋白酶捕获多肽接触，在蛋白酶捕获多肽裂解时形成蛋白酶捕获多肽与集合中的任何蛋白酶或其催化活性部分的共价复合物；
b)从未复合的蛋白酶捕获多肽中分离复合的蛋白酶；
c)分离或选择或鉴别所述复合的蛋白酶；
d)基于选择的蛋白酶产生蛋白酶或蛋白酶的催化活性部分的第二个集合；及
e)通过将第二个蛋白酶或其催化活性部分集合与不同于第一种蛋白酶捕获多肽的第二种蛋白酶捕获多肽接触而重复步骤a)-c)，形成复合物；分离所述复合物；分离、选择或鉴别复合的蛋白酶。
86.权利要求85的方法，其中第一种和第二种蛋白酶捕获多肽被修饰为含有两个不同靶底物裂解序列，从而鉴别的或选择的蛋白酶对于至少两个裂解序列具有特异性和高裂解活性。
87.权利要求85或86的方法，其被重复多次。
88.权利要求85-87任一项的方法，其中将蛋白酶或其催化活性部分的集合与第一种和/或第二种蛋白酶捕获多肽在存在竞争剂的条件下接触。
89.权利要求88的方法，其中所述竞争剂选自人血清、人血浆、细胞或组织提取物、生物学液体如尿液或血液、纯化或部分纯化的野生型形式的蛋白酶捕获多肽及蛋白酶捕获多肽的一或多种特异变体。
90.权利要求1-89任一项的方法，其中所述集合含有修饰的蛋白酶。
91.权利要求90的方法，其中所述修饰是随机修饰或者聚焦修饰或者在靶区域中。
92.权利要求2-91任一项的方法，进一步包括筛选所述复合物对于靶底物的裂解序列的底物特异性。
93.一种组合，其包含：
蛋白酶和/或其催化活性部分的集合；及
至少一种蛋白酶捕获多肽。
94.权利要求93的组合，其中所述蛋白酶捕获多肽选自丝氨酸蛋白酶抑制蛋白、p35家族成员、α-巨球蛋白家族成员或者其修饰形式。
95.权利要求93或94的组合，其中所述蛋白酶和/或其催化活性部分的集合在展示文库中提供。
96.权利要求95的组合，其中所述展示文库是噬菌体展示文库且成员至少展示蛋白酶的催化活性部分。
97.一种试剂盒，其包含权利要求93-96任一项的组合，其中每种成分均单独包装。
98.一种修饰丝氨酸蛋白酶的底物特异性的方法，包括修饰选自基于胰凝乳蛋白酶编号的第30、73、89和155位的一或多个残基。
99.权利要求98的方法，其中所述丝氨酸蛋白酶是uPA。
100.一种修饰的蛋白酶，
其中所述蛋白酶或其催化活性片段是丝氨酸蛋白酶的胰凝乳蛋白酶家族的；及
且在相应于基于胰凝乳蛋白酶编号的第30、73、89和155位的任意位置含有一或多个突变，从而改变底物特异性。
101.权利要求100的修饰的蛋白酶，其中所述蛋白酶是修饰的尿纤溶酶原激活物(u-PA)多肽或其催化活性片段。
102.一种修饰的尿纤溶酶原激活物(u-PA)多肽，其包含相应于SEQ IDNO：433所示u-PA多肽中氨基酸位置的一或多个氨基酸修饰，其中：
所述一或多个氨基酸修饰增加对于靶底物的特异性或者对于靶底物的活性或者这二者，及
所述靶底物参与疾病或病症的病因学。
103.权利要求102的修饰的尿纤溶酶原激活物(u-PA)多肽，其中所述靶底物是VEGFR或者组织纤溶酶原激活物(t-PA)底物。
104.权利要求103的修饰的尿纤溶酶原激活物(u-PA)多肽，其中所述u-PA多肽或其催化活性部分在一定位置含有一或多个修饰，从而改变对于t-PA底物或其序列的底物特异性，所述位置相应于选自SEQ ID NO：433所示u-PA多肽中基于胰凝乳蛋白酶编号的第21、24、30、39、61(A)、80、82、84、89、92、156、158、159和187位位置。
105.权利要求103的修饰的u-PA多肽，其中所述u-PA多肽或其催化活性部分含有选自F21V、I24L、F30V、F30L、T39A、Y61(A)H、E80G、E82E、E84K、I89V、K92E、K156T、T158A、V159A和K187E的一或多个修饰。
106.权利要求105的修饰的u-PA多肽，其中所述u-PA多肽或其催化活性部分含有选自如下的修饰：F21V、I24L、F30V、F30L、F30V/Y61(a)H、F30V/K82E、F30V/K156T、F30V/K82E/V159A、F30V/K82E/T39A/V159A、F30V/K82E/T158A/V159A、F30V/Y61(a)H/K92E、F30V/K82E/V159A/E80G/I89V/K187E和F30V/K82E/V159A/E80G/E84K/I89V/K187E。
107.权利要求103的修饰的尿纤溶酶原激活物(u-PA)多肽，其中所述u-PA多肽或其催化活性部分在一定位置含有一或多个修饰，从而改变了对于VEGFR-2或其序列的底物特异性，所述位置相应于选自SEQ ID NO：433所示u-PA多肽基于胰凝乳蛋白酶编号的第38、72、73、75、132、133、137、138、155、160和217位的位置。
108.权利要求107的修饰的u-PA多肽，其中所述u-PA多肽或其催化活性部分含有选自如下的一或多个修饰：V38D、R72G、L73A、L73P、S75P、F132L、G133D、E137G、I138T、L155P、L155V、L155M、V160A和R217C。
109.权利要求107的修饰的u-PA多肽，其中所述u-PA多肽或其催化活性部分进一步包含在基于胰凝乳蛋白酶编号的第30位和/或第89位的修饰。
110.权利要求109的修饰的u-PA多肽，其中所述u-PA多肽或其催化活性部分中的进一步修饰选自F30I、F30T、F30L、F30V、F30G、F30M和I89V。
111.权利要求110的修饰的u-PA多肽，其中所述u-PA多肽或其催化活性部分含有选自如下的修饰：L73A/I89V、S75P/I89V/I138T、R72G/L155P、F132L/V160A、L73A/I89V/F30T、L73A/I89V/F30L、L73A/I89V/F30V、L73A/I89V/F30G、L73A/I89V/L155V、L73A/I89V/F30M、L73A/I89V/L155M、L73A/I89V/F30L/L155M和L73A/I89V/F30G/L155M。
112.权利要求103的修饰的u-PA多肽，其中所述u-PA多肽或其催化活性部分含有相应于选自SEQ ID NO：433所示u-PA多肽中基于胰凝乳蛋白酶编号的F30I、F30T、F30G和F30M的一或多个突变，从而改变对于VEGFR-2或其序列的底物特异性。
113.权利要求102-112任一项的修饰的u-PA多肽，其中：
所述u-PA多肽与SEQ ID NO：433所示野生型u-PA多肽或其等位基因或物种变体具有80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更高的序列相同性；及
所述多肽包含一或多个修饰。
114.一种修饰的MT-SP1多肽或其催化活性部分，其包含选自SEQ IDNO：253所示MT-SP1多肽中基于胰凝乳蛋白酶编号的如下一或多个氨基酸修饰：D23E、I41F、I41T、L52M、Y60(g)s、T56K、H71R、F93L、N95K、F97Y、F97L、T98P、F99L、A126T、V129D、P131S、I136T、I136V、H143R、T144I、I154V、N164D、T166A、L171F、P173S、F184(a)L、Q192H、S201I、Q209L、Q221(a)L、R230W、F234L和V244G。
115.权利要求114的修饰的MT-SP1多肽，其中所述修饰在具有SEQID NO：505所示氨基酸序列的MT-SP1的催化活性部分中。
116.权利要求114或115的修饰的MT-SP1多肽，其中所述修饰在MT-SP1多肽中，进一步包括相应于SEQ ID NO：253所示MT-SP1多肽中基于胰凝乳蛋白酶编号的C122S的修饰。
117.权利要求116的修饰的MT-SP1多肽，其中所述修饰在具有SEQID NO：507或517所示氨基酸序列的MT-SP1多肽中。
118.权利要求114-117任一项的修饰的MT-SP1多肽，其中所述氨基酸修饰选自I41F、F97Y、L171F和V244G。
119.权利要求114-118任一项的修饰的MT-SP1多肽，其进一步包含相应于SEQ ID NO：253所示MT-SP1多肽中Q175R或D217V修饰的一或多个修饰。
120.权利要求119的修饰的MT-SP1多肽，其中所述氨基酸修饰选自I136T/N164D/T166A/F184(A)L/D217V、I41F、I41F/A126T/V244G、D23E/I41F/T98P/T144I、I41F/L171F/V244G、H143R/Q175R、I41F/L171F、R230W、I41F/I154V/V244G、I141F/L52M/V129D/Q221(A)L、F99L、F97Y/I136V/Q192H/S201I、H71R/P131S/D217V、D217V、T65K/F93L/F97Y/D217V、I41T/P173S/Q209L、F97L/F234L、Q175R、N95K和Y60(G)S。
121.权利要求114-120任一项的修饰的MT-SP1多肽，其中所述修饰增加了对于C2补体蛋白的特异性或者对于C2补体蛋白的活性或者这二者。
122.权利要求114-121任一项的修饰的MT-SP1多肽，其中：
所述MT-SP1多肽与SEQ ID NO：253所示野生型MT-SP1多肽、或者SEQ ID NO：505所示其催化活性部分、或者其等位基因或物种变体具有80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更高的序列相同性；及
所述多肽包含一或多个修饰。
123.一种药物组合物，其包含权利要求102的修饰的u-PA多肽。
124.一种药物组合物，其包含权利要求104-106任一项的修饰的u-PA多肽。
125.一种药物组合物，其包含权利要求107-113任一项的修饰的u-PA多肽。
126.一种药物组合物，其包含权利要求114-122任一项的修饰的MT-SP1多肽。
127.权利要求123-126任一项的药物组合物，其进一步包含药物学可接受的赋形剂。
128.权利要求127的药物组合物，其中所述药物组合物配制为用于全身、口服、经鼻、肺部、表面或者局部给予。
129.一种试剂盒，其包含权利要求123-126任一项的药物组合物、给予所述组合物的装置，及任选地给药说明书。
130.一种核酸分子，其包含编码权利要求102-113任一种修饰的u-PA蛋白酶的核苷酸序列。
131.一种核酸分子，其包含编码权利要求114-122任一种修饰的MT-SP1蛋白酶的核苷酸序列。
132.一种载体，其包含权利要求130或131的核酸分子。
133.一种细胞，其包含权利要求132的载体。
134.一种治疗方法，包括通过给予对象权利要求124的药物组合物进行治疗，其中所述对象患有通过给予t-PA而治疗的疾病或病症。
135.权利要求134的方法，其中所述疾病或病症选自动脉血栓形成、静脉血栓形成和血栓栓塞、缺血性中风、获得性血液凝固障碍、弥散性血管内凝血、细菌感染和牙周炎及神经系统病症。
136.权利要求135的方法，其中所述疾病或病症是阻塞导管的血栓形成。
137.一种治疗方法，包括通过给予对象权利要求125的药物组合物进行治疗，其中所述对象患有由VEGFR-2介导的疾病或病症。
138.权利要求137的方法，其中进行治疗的疾病或病症是血管生成性疾病。
139.权利要求138的方法，其中所述血管生成性疾病选自癌症、黄斑变性、炎症及糖尿病。
140.权利要求139的方法，其进一步包括给予另一种抗肿瘤药物。
141.一种治疗方法，包括通过给予对象权利要求126的药物组合物进行治疗，其中所述对象患有由补体蛋白C2介导的疾病或病症。
142.权利要求141的方法，其中所述疾病或病症选自自身免疫疾病、败血症、类风湿性关节炎(RA)、膜增生性肾小球肾炎(MPGN)、多发性硬化(MS)、重症肌无力(MG)、哮喘、炎症性肠病、免疫复合物(IC)介导的急性炎症性组织损伤、阿尔茨海默病(AD)、缺血-再灌注损伤、黄斑变性、移植器官的排斥反应和Guillan-Barre综合征。
143.权利要求142的方法，其中所述缺血-再灌注损伤是由于选自心肌梗塞(MI)、中风、血管成形术、冠状动脉旁路移植术、心肺转流术(CPB)和血液透析的事件或治疗而导致的。
144.权利要求141的方法，其中所述疾病或病症得自对对象进行的治疗。
145.权利要求144的方法，其中在治疗对象之前给予修饰的MT-SP1。
146.权利要求141的方法，其中给予修饰的MT-SP1是通过将体液或组织样品在体外、离体或者在体内与修饰的MT-SP1接触而完成。
147.权利要求144的方法，其中所述治疗导致补体介导的缺血-再灌注损伤。
148.权利要求147的方法，其中所述治疗是血管成形术或者冠状动脉旁路移植术。
149.权利要求124的药物组合物在治疗通过给予t-PA进行治疗的疾病或病症中的应用。
150.权利要求124的药物组合物在制备通过t-PA治疗的疾病或病症的药物治疗中的应用。
151.权利要求149或150的应用，其中所述疾病或病症选自动脉血栓形成、静脉血栓形成和血栓栓塞、缺血性中风、获得性血液凝固障碍、弥散性血管内凝血、细菌感染和牙周炎及神经系统病症。
152.权利要求149或150的应用，其中所述疾病或病症是阻塞导管的血栓形成。
153.权利要求125的药物组合物在治疗由VEGFR-2介导的疾病或病症中的应用。
154.权利要求125的药物组合物在制备治疗由VEGFR-2介导的疾病或病症的药物中的应用。
155.权利要求153或154的应用，其中进行治疗的疾病或病症是血管形成性疾病。
156.权利要求155的应用，其中所述血管形成性疾病选自癌症、黄斑变性、炎症和糖尿病。
157.权利要求153-156任一项的应用，进一步包括给予另一种抗肿瘤药物。
158.权利要求126的药物组合物在治疗由补体蛋白C2介导的疾病或病症中的应用。
159.权利要求126的药物组合物在配制治疗由补体蛋白C2介导的疾病或病症的药物中的应用。
160.权利要求158或159的应用，其中所述疾病或病症选自自身免疫疾病、败血症、类风湿性关节炎(RA)、膜增生性肾小球肾炎(MPGN)、多发性硬化(MS)、重症肌无力(MG)、哮喘、炎症性肠病、免疫复合物(IC)介导的急性炎症性组织损伤、阿尔茨海默病(AD)、缺血-再灌注损伤、移植器官的排斥反应、黄斑变性和Guillan-Barre综合征。
161.权利要求160的应用，其中所述缺血-再灌注损伤是由于选自心肌梗塞(MI)、中风、血管成形术、冠状动脉旁路移植术、心肺转流术(CPB)和血液透析的事件或治疗而引起的。
162.一种修饰的丝氨酸蛋白酶抑制蛋白多肽，其包含在其反应位点环中相应于P4-P2’位置的任一或多个位置中的修饰，所述修饰具有针对靶底物的裂解序列。
163.权利要求162的修饰的丝氨酸蛋白酶抑制蛋白多肽，其中所述靶底物参与疾病或病症的病因学。
164.权利要求162或163的修饰的丝氨酸蛋白酶抑制蛋白多肽，其中所述靶底物选自VEGFR、补体蛋白或t-PA底物。
165.权利要求164的修饰的丝氨酸蛋白酶抑制蛋白多肽，其中所述靶底物是VEGFR2。
166.权利要求164的修饰的丝氨酸蛋白酶抑制蛋白多肽，其中所述补体蛋白是C2。
167.权利要求162-166任一项的修饰的丝氨酸蛋白酶抑制蛋白多肽，其中所述丝氨酸蛋白酶抑制蛋白是纤溶酶原-激活物抑制剂-1(PAI-1)或者抗凝血酶-3(AT3)。
168.权利要求162-167任一项的修饰的丝氨酸蛋白酶抑制蛋白多肽，其具有SEQ ID NO：497、499、610和611所示任一氨基酸序列。
169.权利要求1-40任一项的方法，其中所述蛋白酶是人激肽释放酶8。
170.一种方法，其包括：
a)将蛋白酶和/或蛋白酶的催化活性部分的集合与蛋白酶捕获多肽接触，在所述蛋白酶捕获多肽由所述蛋白酶或其催化活性部分裂解时，形成含有所述蛋白酶捕获多肽与所述集合中的蛋白酶或其催化活性部分的稳定复合物；及
b)鉴别或选择复合物中的蛋白酶或其催化活性部分，其中所述集合含有两个不同成员。
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102181419A (zh) * | 2011-01-18 | 2011-09-14 | 湖南尤特尔生化有限公司 | 一种里氏木霉蛋白酶及其制备方法和应用 |
| CN111465857A (zh) * | 2017-08-08 | 2020-07-28 | 昆士兰科技大学 | 诊断早期心力衰竭的方法 |
| CN112662694A (zh) * | 2020-12-25 | 2021-04-16 | 康九生物科技(长春)有限公司 | 一种麦芽糖结合蛋白、麦芽糖结合蛋白表达载体、重组工程菌及其应用 |
| CN117969827A (zh) * | 2024-03-28 | 2024-05-03 | 北京大学人民医院 | 抗caspase 14抗体在制备类风湿关节炎补充诊断产品中的应用 |
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| KR20200030063A (ko) * | 2017-06-22 | 2020-03-19 | 카탈리스트 바이오사이언시즈, 인코포레이티드 | 변형된 막 유형 세린 프로테아제 １(ｍｔｓｐ－１) 폴리펩티드 및 사용 방법 |
| WO2020073553A1 (zh) * | 2018-10-10 | 2020-04-16 | 上饶市康可得生物科技有限公司 | 筛选蛋白酶变体的方法以及获得的蛋白酶变体 |
| CN111019925B (zh) * | 2018-10-10 | 2022-08-23 | 上饶市康可得生物科技有限公司 | 筛选蛋白酶变体的方法以及获得的蛋白酶变体 |
| CN111117940B (zh) * | 2019-12-04 | 2022-06-28 | 天津大学 | 一种高产戊二胺的大肠杆菌工程菌与方法 |
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| WO2024055291A1 (zh) * | 2022-09-16 | 2024-03-21 | 中国科学院深圳先进技术研究院 | 高通量筛选sirtuin突变体的方法及sirtuin突变体和其应用 |
| CN116640741A (zh) * | 2023-07-17 | 2023-08-25 | 河北艾欧路生物科技有限责任公司 | 一种从牛血清中提取肌酸激酶的方法 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102181419A (zh) * | 2011-01-18 | 2011-09-14 | 湖南尤特尔生化有限公司 | 一种里氏木霉蛋白酶及其制备方法和应用 |
| CN111465857A (zh) * | 2017-08-08 | 2020-07-28 | 昆士兰科技大学 | 诊断早期心力衰竭的方法 |
| CN112662694A (zh) * | 2020-12-25 | 2021-04-16 | 康九生物科技(长春)有限公司 | 一种麦芽糖结合蛋白、麦芽糖结合蛋白表达载体、重组工程菌及其应用 |
| CN117969827A (zh) * | 2024-03-28 | 2024-05-03 | 北京大学人民医院 | 抗caspase 14抗体在制备类风湿关节炎补充诊断产品中的应用 |
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