CN101094865A - 修饰的生长激素 - Google Patents
修饰的生长激素 Download PDFInfo
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- CN101094865A CN101094865A CNA2005800454376A CN200580045437A CN101094865A CN 101094865 A CN101094865 A CN 101094865A CN A2005800454376 A CNA2005800454376 A CN A2005800454376A CN 200580045437 A CN200580045437 A CN 200580045437A CN 101094865 A CN101094865 A CN 101094865A
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- growth hormone
- modification
- hormone polypeptides
- amino acid
- tethelin
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Abstract
本发明提供了修饰的生长激素多肽、编码修饰的生长激素多肽的核酸分子和产生修饰的生长激素多肽的方法。还提供了用修饰的生长激素多肽进行治疗的方法。
Description
相关申请
要求申请日为2004年11月4日、申请人为Thierry Guyon,Gilles Borrelly,Lila Drittanti,和Manuel Vega、发明名称为“修饰的生长激素”的美国临时申请系列号60/625,652和申请日为2005年8月8日、申请人为Thierry Guyon,Gilles Borrelly,Lila Drittanti,和ManuelVega、发明名称为“修饰的生长激素”的美国临时申请系列号60/706,697的优先权。当允许时,这些申请的内容完整援引加入本文。
本申请与如下申请相关:申请日为2003年9月8日的美国申请系列号10/658,834、公开号为US-2004-0132977-A1和公开的国际PCT申请WO 2004/022747,发明名称为″RATIONAL EVOLUTIONOF CYTOKINES FOR HIGHER STABILITY,THE CYTOKINES ANDENCODING NUCLEIC ACID MOLECULES,″,申请人为Rene Gantier,Thierry Guyon,Manuel Vega and Lila Drittanti。本申请还与如下申请相关:申请日为2003年9月8日的美国申请系列号10/658,355、国际PCT申请WO 2004/022593、发明名称为″RATIONAL DIRECTEDPROTEIN EVOLUTION USING TWO-DIMENSIONAL RATIONALMUTAGENESIS SCANNING,″、申请人为Rene Gantier,ThierryGuyon,Cruz Ramos Hugo,Manuel Vega and Lila Drittanti。当允许时,这些申请的每一个的内容均完整援引加入本文。
发明领域
本发明提供了具有不同于未修饰的野生型蛋白质的性质的修饰的生长激素(GH)蛋白质。还提供了编码这些蛋白质的核酸分子以及使用所述修饰的生长激素蛋白质的治疗和诊断方法。
背景
用于临床用途的治疗性蛋白质的输送是药物学的一项挑战。当这些蛋白质在血流中时,它们在短时间内通过不同的生理学过程而不断地从循环中消除,这些生理学过程涉及代谢和使用蛋白质消除的正常途径的清除,如在肾中的(肾小球)过滤或在血液中的蛋白酶解。后者通常是影响用作经口给予和静脉内或肌内注射的治疗剂的蛋白质的半衰期的限制过程。与蛋白质的这些给予途径相关的问题是公知的并且已经使用了各种策略尝试去解决这些问题。
已经成为许多改善给药和生物同化作用的临床研究和努力的焦点的一种蛋白质家族是细胞因子家族,包括人类生长激素(hGH)。人类生长激素由Harvey Cushing在1912年发现。在1956年,内分泌学家Maurice Raben从人和猴组织中分离了这一激素并且在1958年注射给一侏儒儿童。从那时起,hGH作为一种批准的生物药品的应用经历了两个阶段。在从1958至1985年的第一阶段,hGH得自人垂体提取物。由于热会破坏GH,因此这些提取物不能被加热并从其它生物学污染物如潜在地库贾氏病(Creutzfeldt-Jacob disease)的致病原中完全纯化。在1985年,FDA禁止使用提取的hGH。HGH的药物学应用的第二阶段从重组DNA技术的出现开始,伴随着美国国会在1983年表决的孤儿药法案的实施。基于重组DNA的技术使得可以从哺乳动物或细菌细胞培养物以工业量体外生产重组hGH。
人类GH已被批准用于治疗多种疾病(disease)、病症(disorder)和状况(condition)如儿童生长激素缺陷、成人生长激素缺陷、特纳氏综合征(Turner’s Syndrome)、慢性肾功能不全、与AIDS相关的恶病质及短肠综合征。因此,生长激素和其它细胞因子一样是重要的治疗剂。天然发生的变体通常具有不希望的副作用,以及上述给药、生物利用度和短半衰期等问题。因此,需要改善GH的性质以利于其用作生物治疗剂。因此,在本文的各种目的中,一个目的是提供具有改善的治疗性质的GH变体。
概述
本文提供了修饰的生长激素(GH),它们展示出与未修饰的生长激素相比增加的蛋白质稳定性。生长激素的用途对于人类和其它动物是熟知的。GH的目前应用由于其在血流中和储存条件下的不稳定性而需要频繁和重复应用。本文提供的修饰的GH是GH的突变变体,它们展示改善的稳定性,由此具有增加的蛋白质半衰期,包括在血流和/或储存条件下的增加的稳定性。这种改善的稳定性包括通过对蛋白酶的抗性和/或增加的热耐受性而评价的稳定性。
本文提供的修饰的生长激素多肽展示出增加的蛋白质稳定性。所提供的变体是与未修饰的GH多肽相比在一或多个氨基酸位置被修饰的那些变体。本文提供的修饰的生长激素多肽包括前体形式和成熟形式、较长形式和较短形式;修饰是相对于成熟形式而描述。本领域技术人员能容易地确定一特定多肽上的相应位置,如根据未改变的残基的序列对比而确定。
本发明提供了修饰的生长激素多肽,它们与未修饰的生长激素相比在相应于成熟人类生长激素的氨基酸位置1-55、57、58、60-63、67-87、89-91、93、95-100、102-128、131-132、135-139、141、142、144、148-182、184、185以及187-191的任何位置含有1-5个氨基酸置换,其中成熟人类生长激素含有SEQ IDNO:1所示的氨基酸残基序列。应理解的是序列为SEQ ID NO:1的多肽有等位基因变体、物种变体和同种型,这些多肽也可以在相应于本文举例示出的多肽的位点被修饰。与SEQ ID NO:1或SEQ IDNO:712(示出包括信号序列的前体形式)的未修饰的生长激素相比,修饰的生长激素显示增加的蛋白质稳定性。
本发明提供了修饰的生长激素多肽,它们与未修饰的生长激素相比含有一或多个单氨基酸残基置换,所述单氨基酸置换发生在相应于成熟人类生长激素的氨基酸位置1-12、14-26、29-53、57、58、60-63、67-78、80-84、86、87、89、91、93、95-100、102-113、115-128、131、132、135-139、141、142、144、148-160、162-182、185以及187-191的任何一或多个位置,其中成熟人类生长激素具有SEQ ID NO:1所示的氨基酸残基序列,并且与含有SEQ ID NO:1或SEQ ID NO:712的氨基酸序列的未修饰的生长激素相比,修饰的生长激素显示增加的蛋白质稳定性。
本发明提供了与未修饰的生长激素相比具有一或多个单氨基酸置换的修饰的生长激素,其中所述置换位置不是相应于SEQ ID NO:1所示的成熟人类生长激素的位置13、27、28、54-56、59、64-66、79、85、88、90、92、94、101、114、129、130、133、134、140、143、145-147、161、183、184和186的位置。在一个特定的实施方案中,修饰的生长激素显示与未修饰的生长激素相比增加的蛋白质稳定性。增加的蛋白质稳定性可以体现为增加的血清半衰期。
本发明提供了修饰的生长激素,其与未修饰的生长激素相比在相应于SEQ ID NO:1、712或713所示的成熟人类生长激素的氨基酸位置1-55、57、58、60-63、67-87、89-91、93、95-100、102-128、131、132、135-139、141、142、144、148-182、184、185以及187-191的任何位置具有一或多个单氨基酸置换。在本发明提供的一个特定实施方案中,修饰的生长激素显示与未修饰的生长激素相比增加的蛋白质稳定性,其中如果位置9被置换,则置换氨基酸不是脯氨酸,如果位置14被置换,则置换氨基酸不是丝氨酸,如果位置13或27被置换,则置换氨基酸不是缬氨酸,如果位置28被置换,则置换氨基酸不是苯丙氨酸,如果位置54被置换,则置换氨基酸不是酪氨酸,如果位置55,79,85或184被置换,则置换氨基酸不是丙氨酸,如果位置90被置换,则置换氨基酸不是异亮氨酸,如果位置114或161被置换,则置换氨基酸不是甲硫氨酸,如果位置120或126被置换,则置换氨基酸不是精氨酸。
本发明提供的修饰的生长激素包括在如下位置的一或多个位置的氨基酸置换:成熟人类生长激素多肽的1、2、5、6、8、9、10、11、14、15、16、19、20、23、25、26、28、30、31、32、33、35、37、38、39、41、42、44、45、48、52、54、61、70、73、74、75、76、77、80、81、82、86、87、89、93、97、103、107、111、112、113、114、115、116、117、118、119、124、125、127、128、139、153、154、156、157、158、160、162、163、164、166、167、168、169、170、171、172、174、176、177、178和191。在一个实施方案中,所述位置包括成熟人类生长激素的F1、P2、P5、L6、R8、L9、F10、D11、M14、L15、R16、R19、L20、L23、F25、D26、Y28、E30、F31、E32、E33、Y35、P37、K38、E39、K41、Y42、F44、L45、P48、L52、F54、P61、K70、L73、E74、L75、L76、R77、L80、L81、L82、W86、L87、P89、L93、F97、Y103、D107、Y111、D112、L113、L114、K115、D116、L117、E118、E119、L124、M125、R127、L128、F139、D153、D154、L156、L157、K158、Y160、L162、L163、Y164、F166、R167、K168、D169、M170、D171、K172、E174、F176、L177、R178和F191。在一个实施方案中,位置被如下置换:由H或Q置换R,由H、Q或N置换E,由Q或N置换K,由N或Q置换D,由I或V置换M,由A或S置换P,由I或H置换Y,由I或V置换F,由H或S置换W,由I或V置换L。例如,这类置换包括F1I(即在相应于成熟人类生长激素(例如SEQ ID NO:1或712)的氨基酸位置1的位置由I置换F),F1V,P2A,P2S,P5A,P5S,L6I,L6V,R8H,R8Q,L9I,L9V,F10I,F10V,D11N,D11Q,M14I,M14V,L15I,L15V,R16H,R16Q,R19H,R19Q,L20I,L20V,L23I,L23V,F25I,F25V,D26N,D26Q,Y28H,Y28I,E30Q,E30H,E30N,F31I,F31V,E32Q,E32H,E32N,E33Q,E33H,E33N,Y35H,Y35I,P37A,P37S,K38N,K38Q,E39Q,E39H,E39N,K41N,K41Q,Y42H,Y42I,F44I,F44V,L45I,L45V,P48A,P48S,L52I,L52V,F54I,F54V,P61A,P61S,K70N,K70Q,L73I,L73V,E74Q,E74H,E74N,L75I,L75V,L76I,L76V,R77H,R77Q,L80I,L80V,L81I,L81V,L82I,L82V,W86H,W86S,L87I,L87V,P89A,P89S,L93I,L93V,F97I,F97V,Y103H,Y103I,D107N,D107Q,Y111H,Y111I,D112N,D112Q,L113I,L113V,L114I,L114V,K115N,K115Q,D116N,D116Q,L117I,L117V,E118Q,E118H,E118N,E119Q,E119H,E119N,L124I,L124V,M125I,M125V,R127H,R127Q,L128I,L128V,F139I,F139V,D153N,D153Q,D154N,D154Q,L156I,L156V,L157I,L157V,K158N,K158Q,Y160H,Y160I,L162I,L162V,L163I,L163V,Y164H,Y164I,F166I,F166V,R167H,R167Q,K168N,K168Q,D169N,D169Q,M170I,M170V,D171N,D171Q,K172N,K172Q,E174Q,E174H,E174N,F176I,F176V,L177I,L177V,R178H,R178Q,F191I和F191V。
在另一个实施方案中,本发明提供的修饰的生长激素包括在如下一或多个位置的氨基酸置换:含有SEQ ID NO:1的氨基酸残基序列的成熟人类生长激素多肽的位置1、2、5、9、11、14、16、23、26、38、41、42、73、74、81、87、111、112、116、119、124、125、153、156、157、158、162、166、167、168、169、171、172、174、177、178和191。在一个实施方案中,所述位置包括成熟人类生长激素多肽的F1、P2、P5、L9、D11、M14、R16、L23、D26、K38、K41、Y42、L73、E74、L81、L87、Y111、D112、D116、E119、L124、M125、D153、L156、L157、K158、L162、F166、R167、K168、D169、D171、K172、E174、L177、R178和F191。例如,这些置换包括但不限于F1I(即在相应于成熟生长激素的氨基酸位置1的位置由I置换F),P2A,P5S,L9V,D11N,M14V,R16H,L23I,L23V,D26N,K38N,K41Q,Y42H,Y42I,L73V,E74N,L81V,L87V,Y111I,D112N,D116Q,E119Q,L124V,M125I,M125V,D153N,L156I,L157I,K158N,L162I,F166I,R167H,R167Q,K168N,K168Q,D169Q,D171N,D171Q,K172Q,E174Q,E174N,E174H,L177V,L177I,R178Q和F191I。在一个特定的实施方案中,在具有SEQ ID NO:1的氨基酸序列的成熟人类生长激素多肽中,如果位置F1由I或V置换,则位置P2由A置换。
在另一个实施方案中,本发明提供的修饰的生长激素包括在一或多个如下位置的氨基酸置换:含有SEQ ID NO:1的氨基酸残基序列的成熟人类生长激素的位置6、9、13、15、17、20、23、24、105、110、113、114、117、121、124和128。在一个实施方案中,所述位置包括成熟人类生长激素的L6、L9、A13、L15、A17、L20、L23、A24、A105、V110、L113、L114、L117、I121、L124和L128。在一个实施方案中,位置用E、D、K、R、N、Q、S和/或T置换。例如,这类置换包括L6E(即在相应于成熟生长激素的氨基酸位置6的位置由E置换L)、L6D、L6K、L6R、L6N、L6Q、L6S、L6T、L9E、L9D、L9K、L9R、L9N、L9Q、L9S、L9T、A13E、A13D、A13K、A13R、A13N、A13Q、A13S、A13T、L15E、L15D、L15K、L15R、L15N、L15Q、L15S、L15T、A17E、A17D、A17K、A17R、A17N、A17Q、A17S、A17T、L20E、L20D、L20K、L20R、L20N、L20Q、L20S、L20T、L23E、L23D、L23K、L23R、L23N、L23Q、L23S、L23T、A24E、A24D、A24K、A24R、A24N、A24Q、A24S、A24T、A105E、A105D、A105K、A105R、A105N、A105Q、A105S、A105T、V110E、V110D、V110K、V110R、V110N、V110Q、V110S、V110T、L113E、L113D、L113K、L113R、L113N、L113Q、L113S、L113T、L114E、L114D、L114K、L114R、L114N、L114Q、L114S、L114T、L117E、L117D、L117K、L117R、L117N、L117Q、L117S、L117T、I121E、I121D、I121K、I121R、I121N、I121Q、I121S、I121T、L124E、L124D、L124K、L124R、L124N、L124Q、L124S、L124T、L128E、L128D、L128K、L128R、L128Q、L128N、L128S和L128T。
本发明提供如上所述的任何修饰的生长激素,进一步具有在含有SEQ ID NO:1的氨基酸残基序列的成熟人类生长激素的位置6、9、13、15、17、20、23、24、105、110、113、114、117、121、124和/或128的一或多个单氨基酸置换,其中如果位置13被置换,则置换氨基酸不是缬氨酸,如果位置114被置换,则置换氨基酸不是甲硫氨酸。在一个实施方案中,所述位置包括L6、L9、A13、L15、A17、L20、L23、A24、A105、V110、L113、L114、L117、I121、L124和L128。在一个实施方案中,位置用E,D,K,R,N,Q,S和/或T置换。例如,这类置换包括L6E、L6D、L6K、L6R、L6N、L6Q、L6S、L6T、L9E、L9D、L9K、L9R、L9N、L9Q、L9S、L9T、A13E、A13D、A13K、A13R、A13N、A13Q、A13S、A13T、L15E、L15D、L15K、L15R、L15Q、L15N、L15S、L15T、A17E、A17D、A17K、A17R、A17N、A17Q、A17S、A17T、L20E、L20D、L20K、L20R、L20N、L20Q、L20S、L20T、L23E、L23D、L23K、L23R、L23N、L23Q、L23S、L23T、A24E、A24D、A24K、A24R、A24N、A24Q、A24S、A24T、A105E、A105D、A105K、A105R、A105N、A105Q、A105S、A105T、V110E、V110D、V110K、V110R、V110N、V110Q、V110S、V110T、L113E、L113D、L113K、L113R、L113N、L113Q、L113S、L113T、L114E、L114D、L114K、L114R、L114N、L114Q、L114S、L114T、L117E、L117D、L117K、L117R、L117N、L117Q、L117S、L117T、I121E、I121D、I121K、I121R、I121N、I121Q、I121S、I121T、L124E、L124D、L124K、L124R、L124N、L124Q、L124S、L124T、L128E、L128D、L128K、L128R、L128Q、L128N、L128S和L128T。
本发明提供了如上所述的任何修饰的生长激素,其中与未修饰的生长激素相比被置换的位置数量是2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19或20个。在一个实施方案中,本发明提供的修饰的生长激素包括在一或多个如下位置的氨基酸置换:含有SEQ ID NO:1的氨基酸序列的成熟人类生长激素的位置56、59、64、65、66、88、92、94、101、129、130、133、134、140、143、145、146、147,183和186。在一个实施方案中,所述位置包括E56、P59、R64、E65、E66、E88、F92、R94、L101、E129、D130、P133、R134、K140、Y143、K145、F146、D147、R183和E186。在一个实施方案中,位置被如下置换:由Q、N或H置换E,由S或A置换P,由H或Q置换R,由I或V置换L或F,由Q或N置换K或D,由H或I置换Y。例如,这类置换包括E56Q、E56N、E56H、P59S、P59A、R64H、R64Q、E65Q、E65N、E65H、E66Q、E66N、E66H、E88Q、E88N、E88H、F92I、F92V、R94H、R94Q、L101V、L101I、E129Q、E129N、E129H、D130Q、D130N、P133S、P133A、R134H、R134Q、K140Q、K140N、Y143H、Y143I、K145Q、K145N、F146I、F146V、D147Q、D147N、R183H、R183Q、E186Q、E186N和E186H。
在一个实施方案中,上述修饰的生长激素是人类生长激素(hGH)。在一个特定的实施方案中,氨基酸置换在人垂体生长激素或人胎盘生长激素中进行。或者,上述修饰的生长激素是非人生长激素。
本发明提供了长度为191、192、193、194、195、196、197、198、199、200、201、202、203、204、205、206、207、208、209、210、211、212、21 3、214、215、216、217、218、219、220、221或222个氨基酸的修饰的生长激素。在另一个实施方案中,本发明提供了长度为180、181、182、183、184、185、186、187、188、189或190个氨基酸的修饰的生长激素。
在一个实施方案中,氨基酸置换在SEQ ID NO:1的生长激素序列中进行。在另一个实施方案中,氨基酸置换在SEQ ID NO:712的前体生长激素序列中进行。本发明提供了修饰的生长激素,其中一或多个氨基酸置换是天然氨基酸、非天然氨基酸或天然和非天然氨基酸的组合。
本发明提供了修饰的成熟人类生长激素多肽和修饰的前体人类生长激素多肽。
在某些实施方案中,修饰的生长激素是裸多肽链。
本发明提供了修饰的生长激素,其中所述生长激素是PEG化的、白蛋白化的(albuminated)、糖基化的或用另一种化学方法处理的。
本发明提供了进一步具有一或多个假野生型突变(pseudo-wildtype mutation)的修饰的生长激素。在一个特定的实施方案中,所述假野生型突变可以是未修饰的生长激素的氨基酸残基的插入、缺失、置换或其组合。
在一个方面中,修饰的生长激素显示表现为对蛋白酶解的增加的抗性的增加的稳定性。在某些实施方案中,对蛋白酶解的增加的抗性发生在血清、血液、唾液、消化液中或体外暴露于蛋白酶的情况中。在一个实施方案中,当修饰的生长激素被口服给药或存在于消化道中时,其显示对蛋白酶解的增加的抗性。蛋白酶可以是例如明胶酶A,明胶酶B,胰蛋白酶,胰蛋白酶(Arg blocked),胰蛋白酶(Lysblocked),梭菌蛋白酶,内切蛋白酶(endoproteinase)Asp-N,胰凝乳蛋白酶,溴化氰,iodozobenzoate,Myxobacter P.,Armillaria,鲁米那胃蛋白酶,微绒毛内肽酶,二肽基肽酶,肠肽酶和水解酶中一或多种。在一个实施方案中,修饰的多肽对蛋白酶解的抗性增加至少5%、10%、15%、20%、25%、30%、35%、40%、45%、50%、55%、60%、65%、70%、75%、80%、85%、90%、95%、96%、97%、98%、99%或100%。
在蛋白酶是明胶酶B的一个实施方案中,修饰的多肽对在氨基酸序列Met-Ser-Tyr-Asn处的裂解有抗性。在一个方面,修饰的多肽可以在氨基酸序列Met-Ser-Tyr-Asn后面被裂解。在另一方面,多肽可以在Met、Ser、Tyr、Asn每一残基或其组合被修饰以便赋予修饰的肽以对明胶酶B的蛋白酶解更高的抗性。在一个实施方案中,修饰的多肽对明胶酶B在Met、Ser、Tyr、Asn每一残基或其组合的蛋白酶解的抗性增加了至少5%、10%、15%、20%、25%、30%、35%、40%、45%、50%、55%、60%、65%、70%、75%、80%、85%、90%、95%、96%、97%、98%、99%或100%。在一个实施方案中,对蛋白酶的抗性可以通过本文描述的任何测定经验测试。
在一个方面,修饰的生长激素显示表现为增加的热耐受性的增加的稳定性。在一个实施方案中,修饰的生长激素在约20℃至约45℃的温度有增加的热耐受性。在一个实施方案中,修饰的生长激素在对象的体温有增加的热耐受性。在一个特定的实施方案中,修饰的生长激素在约37℃的温度有增加的热耐受性。
在一个方面,修饰的生长激素显示表现为增加的体内或体外半衰期的增加的稳定性。在一个实施方案中,增加的稳定性表现为当被给予对象时增加的半衰期。与未修饰的生长激素的半衰期相比,修饰的生长激素的半衰期(血清稳定性)可以被增加如10%、至少20%、至少30%、至少40%、至少50%、至少60%、至少70%、至少80%、至少90%、至少100%、至少150%、至少200%、至少250%、至少300%、至少350%、至少400%、至少450%以及至少500%或更高。在另一个实施方案中,与未修饰的生长激素的半衰期相比,修饰的生长激素的半衰期(血清稳定性)可以被增加如至少6倍、7倍、8倍、9倍、10倍、20倍、30倍、40倍、50倍、60倍、70倍、80倍、90倍、100倍、200倍、300倍、400倍、500倍、600倍、700倍、800倍、900倍和1000倍或更多倍。
在一个方面,修饰的生长激素显示与未修饰的生长激素相比增加的生物学活性。在另一方面,修饰的生长激素显示与未修饰的生长激素相比降低的生物学活性。在一个实施方案中,生物学活性通过体外测量细胞增殖而评价。在某些实施方案中,修饰的生长激素与生长激素受体结合。
本发明提供了与未修饰的生长激素相比显示增加的对蛋白酶解的抗性和显示降低的热耐受性的修饰的生长激素。
本发明提供了与未修饰的生长激素相比显示增加的热耐受性和显示降低的对蛋白酶解的抗性的修饰的生长激素。
本发明提供了具有2个或更多个本文描述的任何修饰的生长激素的文库。
本发明提供了含有编码本文所述的修饰的生长激素的核苷酸序列的核酸分子。本发明提供了具有多个本文描述的分子的核酸分子的文库。
本发明提供了含有本文描述的核酸分子的载体。本发明提供了携带本文描述的核酸分子或载体的细胞。在一个实施方案中,所述细胞是真核细胞、原核细胞、昆虫细胞、哺乳动物细胞等。本发明还提供了具有多个载体的文库。
本发明提供了表达修饰的生长激素的方法,其具有如下步骤:i)将核酸或载体导入细胞,和ii)在编码的修饰的生长激素被表达的条件下培养细胞。在一个实施方案中,载体是在真核细胞、原核细胞、昆虫细胞、哺乳动物细胞、植物细胞等中。在一个实施方案中,修饰的生长激素是糖基化的。在一个实施方案中,细胞是真核细胞。
本发明提供了产生具有进化的(evolved)预定性质或活性的修饰的靶蛋白的方法,所述方法包括如下步骤:a)在靶蛋白上选择一或多个在氨基酸置换时适于提供进化的预定性质或活性的is-HIT靶氨基酸;其中所述is-HIT靶氨基酸通过鉴别进化中的靶蛋白与具有预定性质或活性的本文所述的修饰的生长激素之间的结构同源位置而确定;b)用适于提供进化的预定性质或活性的置换氨基酸来置换每个靶氨基酸以形成候选先导蛋白质,其中在每个靶蛋白上仅发生一个氨基酸置换;c)在包含在可寻址阵列中的细胞中从核酸分子表达每个候选先导蛋白质;以及d)测定每个候选先导蛋白质以鉴别一或多种具有不同于未修饰的蛋白质的预定性质或活性的蛋白质,由此鉴别是先导物(LEAD)的进化的靶蛋白。在一个实施方案中,所述方法还包括如下步骤:e)比较进化中的蛋白质与修饰的生长激素的三维结构以鉴别它们主链中的高重合,这些区域被称为同源区域;和f)鉴别进化中的蛋白质上的相应于修饰的生长激素的结构同源区域内的结构相关is-HIT氨基酸位置的is-HIT结构同源位点。在一个实施方案中,预定性质或活性是蛋白质稳定性、体内蛋白质半衰期、热耐受性或对蛋白酶解的抗性。本发明提供了由上述方法产生的修饰的细胞因子。
本发明提供了由本文所述的任何方法产生的修饰的生长激素多肽。
本发明提供了具有SEQ ID NO:2-69、75、76、85-107、111、112、115、116、119、120、123-154、164、165、176-216、222-351所示的氨基酸序列的修饰的生长激素或其生物学活性部分。
本发明提供了包括本文所示的任何修饰的生长激素多肽的药物组合物。药物组合物可进一步包括药物学可接受的赋形剂如结合剂、填充剂、润滑剂、崩解剂和湿润剂。
药物组合物可配制用于口服、鼻和肺给药。在一个特定的实施方案中,药物组合物配制为用于口服给药。药物组合物中的修饰的生长激素在药物制剂中显示在如下条件下的增加的半衰期,所述条件选自暴露于唾液、暴露于胃肠道中的蛋白酶和暴露于与野生型细胞因子相比的低pH条件。
本发明的药物组合物中提供的是修饰的生长激素多肽,其中修饰包括去除蛋白酶消化位点或增加蛋白质结构的稳定性。药物组合物中的修饰的生长激素显示增加的蛋白质半衰期或胃肠道中的生物利用度。与野生型蛋白质的半衰期相比,蛋白质半衰期(血清稳定性)可以被增加至少10%、至少20%、至少30%、至少40%、至少50%、至少50%、至少60%、至少70%、至少80%、至少90%、至少100%、至少150%、至少200%、至少250%、至少300%、至少350%、至少400%、至少450%或500%或更高。或者,与野生型蛋白质相比,蛋白质半衰期可以被增加至少6倍、至少7倍、至少8倍、至少9倍、至少10倍、至少20倍、至少30倍、至少40倍、至少50倍、至少60倍、至少70倍、至少80倍、至少90倍、至少100倍、至少200倍、至少300倍、至少400倍、至少500倍、至少600倍、至少700倍、至少800倍、至少900倍或至少1000倍或更多倍。
在本发明中,蛋白质半衰期(血清稳定性)在存在一或多种蛋白酶时被增加。在一个实施方案中,修饰的生长激素的生物学活性与野生型生长激素相比被增加。
蛋白质半衰期可以在暴露于一或多种条件后被增加,所述一或多种条件包括但不限于暴露于唾液、暴露于胃肠道中的蛋白酶和暴露于低pH条件(例如胃)。
本发明提供了未经使用蛋白酶抑制剂如Bowman-Birk抑制剂、缀合的Bowman-Birk抑制剂、抑酶肽和卡莫他特(camostat)而制备的药物组合物。
本发明提供了例如液体、丸剂、片剂或胶囊剂形式的配制用于口服给药的药物组合物。在一个实施方案中,丸剂或片剂是可嚼的。在一个实施方案中,丸剂或片剂当暴露于舌上或口腔中的唾液时被溶解。在一个实施方案中,胶囊剂中的药物组合物是液体形式。在其中药物组合物是液体的实施方案中,液体可以是例如溶液、糖浆或悬浮液。
本发明提供了配制用于控制释放修饰的生长激素多肽的药物组合物。在一个例子中,所述药物组合物是片剂或锭剂。锭剂将修饰的生长激素输送至口腔粘膜、喉粘膜或胃肠道。另外,锭剂可以用赋形剂配制,如在无水结晶麦芽糖和硬脂酸镁中。
本发明提供了未加保护性化合物而配制的药物组合物。在一个实施方案中,修饰的生长激素展示对肠道蛋白酶的抗性。
药物组合物可以进一步用一或多种药物学可接受的添加剂如悬浮剂、乳化剂、非水载体和/或防腐剂而配制。
本发明提供了由编码本文所述的任一种修饰的生长激素多肽的核酸分子或具有编码本文所述的任一种修饰的生长激素多肽的核酸分子的载体和药物学可接受的赋形剂组成的药物组合物。
本发明提供了通过给予本文所述的任一种药物组合物而治疗患有生长激素介导的疾病或状况的对象的方法。
本发明提供了通过给予本文所述的任一种药物组合物而治疗患有生长激素介导的疾病或状况的对象的方法。在一个实施方案中,生长激素介导的疾病或状况包括但不限于生长缺陷疾病、AIDS消瘦、衰老、HIV感染对象的受损的免疫功能、分解代谢疾病、手术恢复、充血性心肌病、肝移植、肝切除术后肝再生、慢性肾功能衰竭、肾性骨营养不良、骨质疏松、软骨发育不全/季肋发育不全、骨骼发育异常、慢性炎性或营养性疾病如克罗恩氏病、短肠综合征、少年慢性关节炎、囊性纤维化、男性不育症、X连锁低磷血症性佝偻病、唐氏综合症、脊柱裂、努南综合征、肥胖症、肌肉强度受损以及纤维肌痛。在一个特定的实施方案中,生长缺陷疾病可以选自特纳氏综合征、子宫内生长迟缓、特发性身材矮小症、Prader Willi综合征和地中海贫血。在一个实施方案中,对象的症状被改善或消除。
本发明提供了制品,包括但不限于包装材料和含在包装材料内的本文描述的修饰的生长激素多肽的药物组合物。在一个特定的实施方案中,包装在制品内的药物组合物对于治疗生长激素介导的疾病或病症是有效的,并且包装材料包括指示修饰的生长激素用于治疗生长激素介导的疾病或病症的标签。
本发明提供了本发明的修饰的生长激素多肽的药物组合物在制备用于治疗患有由给予生长激素而治疗的疾病或状况的对象的药物中的应用。在所述应用的一个实施方案中,所述生长激素疾病或状况包括但不限于生长缺陷疾病、AIDS消瘦、衰老、HIV感染对象的受损的免疫功能、分解代谢疾病、手术恢复、充血性心肌病、肝移植、肝切除术后肝再生、慢性肾功能衰竭、肾性骨营养不良、骨质疏松、软骨发育不全/季肋发育不全、骨骼发育异常、慢性炎性或营养性疾病如克罗恩氏病、短肠综合征、少年慢性关节炎、囊性纤维化、男性不育症、X连锁低磷血症性佝偻病、唐氏综合症、脊柱裂、努南综合征、肥胖症、肌肉强度受损以及纤维肌痛。在所述应用的一个特定实施方案中,生长缺陷疾病选自特纳氏综合征、子宫内生长迟缓、特发性身材矮小症、Prader Willi综合征以及地中海贫血。
本发明提供了试剂盒,其包括本文所述的修饰的生长激素多肽的药物组合物、用于给予修饰的生长激素多肽的装置以及任选地包括给药说明。
本发明提供了本发明的任一种修饰的生长激素多肽在配制用于治疗患有由给予生长激素而治疗的疾病或状况的对象的药物中的应用。本发明提供了本发明的药物组合物在配制用于治疗患有由给予生长激素而治疗的疾病或状况的对象的药物中的应用。在一个实施方案中,所述疾病或状况是下述任一种:生长缺陷疾病、AIDS消瘦、衰老、HIV感染对象的受损的免疫功能、分解代谢疾病、手术恢复、充血性心肌病、肝移植、肝切除术后肝再生、慢性肾功能衰竭、肾性骨营养不良、骨质疏松、软骨发育不全/季肋发育不全、骨骼发育异常、慢性炎性或营养性疾病如克罗恩氏病、短肠综合征、青少年慢性关节炎、囊性纤维化、男性不育症、X连锁低磷血症性佝偻病、唐氏综合症、脊柱裂、努南综合征、肥胖症、肌肉强度受损以及纤维肌痛。在一个方面,生长缺陷疾病是特纳氏综合征、子宫内生长迟缓、特发性身材矮小症、Prader Willi综合征以及地中海贫血中的任一种。
附图简述
图1显示了成熟野生型人类生长激素的序列,对蛋白酶解敏感的氨基酸以黑体表示。
图2描述了用于鉴别置换is-HIT的候选置换氨基酸的“PointAccepted Mutation”(PAM250)矩阵。原始氨基酸在水平轴上示出,置换氨基酸在垂直轴上示出。赋予相同残基的值以灰色框表示。矩阵中的最高值以黑色框表示并相应于两个残基之间最高的取代发生率。
图3显示了例举的修饰的生长激素变体的活性曲线。
详细描述
提纲
A. 定义
B. 生长激素
1. 生长激素结构
2. 与生长激素受体的相互作用
3. 作为生物药物的生长激素
C. 用于修饰生长激素的举例性方法
1. 1D扫描(“合理诱变(Rational Mutagenesis)”)
2. 3D扫描
3. 2D-扫描(限制合理诱变(restricted rational mutagenesis))
a. 鉴别in-silico HIT
b. 鉴别置换氨基酸
c. 构建修饰的多肽和生物学测定
D. 修饰的生长激素多肽
1. 通过去除蛋白酶解位点而实现的对蛋白酶解的增加的抗性
a. 通过去除蛋白酶解位点而修饰的生长激素多肽的性质
b. 通过去除蛋白酶解位点而修饰的生长激素多肽的产生
c. 另外的修饰的生长激素多肽
d. 具有对蛋白酶解增加的抗性的生长激素变体的评估
2. 增加的热耐受性
a. 耐热的修饰的生长激素多肽的性质
i. 分子内键的产生
ii. 增加螺旋之间的极性相互作用
b. 耐热的修饰的生长激素多肽的评估
3. Super-LEAD和另外的生长激素修饰
E. 修饰的生长激素多肽的生产
1. 表达系统
a. 原核表达
b. 酵母
c. 昆虫和昆虫细胞
d. 哺乳动物细胞
e. 植物
2. 纯化
3. 融合蛋白
4. 多肽修饰
5. 核苷酸序列
F. 评价修饰的生长激素活性
1. 体外测定
2. 非人动物模型
3. 临床测定
G. 配制/给药
H. 治疗用途
1. 生长缺陷
2. 恶病质
3. 抗衰老
4. 肾性骨营养不良
5. 囊性纤维化
6. 其它状况
I. 制品(Articles of manufacture)和药盒
J. 实施例
A.定义
除非特别指明,否则本文所用的所有技术和科学术语具有与本发明所属技术领域的技术人员通常理解的相同的含义。除非特别指明,否则本文引用的所有专利、专利申请、公开的申请和出版物、GenBank序列、网址和其它公开的材料均以全文引入作为参考。当本文的术语存在多个定义时,以在本部分的定义为主。当引用URL或其它这类标识符或地址时,应理解这类标识符可以改变并且互联网上的信息可以变化不定,但是通过搜索互联网可以发现等价信息,其中的引用是这些信息的可得性和公共传播的证据。
如本文所用,“生长激素”多肽(本文中也称为GH)是指任何GH多肽(蛋白质),包括但不限于重组产生的多肽、合成产生的多肽和从包括但不限于垂体和胎盘组织的组织的细胞提取的GH。GH多肽包括具有信号序列的前体生长激素多肽和成熟生长激素多肽。GH多肽包括来自不同物种包括但不限于人和非人起源的动物的相关多肽。GH多肽氨基酸序列可以含有变化数目的氨基酸残基。例如,GH多肽可以具有191、192、193、194、195、196、197、198、199、200、201、202、203、204、205、206、207、208、209、210、211、212、213、214、215、216、217、218、219、220、221和222或更多个氨基酸。GH多肽也可以短于191个氨基酸,如180、181、182、183、184、185、186、187、188、189或190个氨基酸,其中所述多肽保留成熟GH多肽的活性。人类GH(hGH)包括GH、个体间或物种间等位基因变体同种型、来自核酸的合成分子、分离自人类组织和细胞的蛋白质、分离自非人组织和细胞的蛋白质,以及它们的修饰形式。举例性的hGH序列如SEQ ID NO:1,712和713所示。一种举例性的成熟hGH多肽的氨基酸序列见SEQID NO:1。举例性的前体生长激素多肽的氨基酸序列见SEQ ID NO:712和713。这些前体多肽的信号肽长度为26个氨基酸。除了个体间和物种间等位基因变异之外,还有分离自胎盘的几种同种型(SEQ IDNO:713)和分离自脑垂体的同种型(SEQ ID NO:712)。人GH还包括长度足以有功能活性的GH片段。确定GH修饰形式的功能性的细胞因子测定对于本领域普通技术人员是已知的。任选地,不具有如下突变:与SEQ ID NOS:1,712和713的氨基酸序列相比的P6S,H22D,G34N或A127V。非人来源的举例性GH多肽包括但不限于牛、羊、猪、大鼠、兔、马、灵长类和禽类GH多肽。举例性的非人生长激素序列见SEQ ID NOS:714和715。非人GH包括GH、等位基因变体同种型、来自核酸的合成分子、分离自非人组织和细胞的蛋白质,以及它们的修饰形式。非人GH还包括长度足以有功能活性的GH片段。确定GH修饰形式的功能性的细胞因子测定对于本领域普通技术人员是已知的。
如本文所用,GH多肽(蛋白质)的“活性”或“性质”是指由GH蛋白展示的可以被评估的任何活性或性质。这类活性包括但不限于对蛋白酶解的抗性、热耐受性、增加的半衰期和细胞增殖活性。
如本文所用,“对蛋白酶解的抗性”是指与用相同蛋白酶在相同条件下对未修饰的多肽的裂解相比,修饰多肽的靶氨基酸残基的任何量的降低的裂解。展示对蛋白酶解的增加的抗性的修饰的多肽展示比未修饰的多肽高例如1%、2%、3%、4%、5%、6%、7%、8%、9%、10%、…20%、…30%、…40%、…50%、…60%、…、70%、…80%、…90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%的对蛋白酶解的抗性。
“蛋白酶(Proteases)”、“蛋白酶(proteinases)”或“肽酶”可互换使用,是指催化共价肽键水解的酶。对于“丝氨酸蛋白酶”,机制是基于丝氨酸对靶向的肽键的亲核攻击。半胱氨酸、苏氨酸或与天冬氨酸盐或金属结合的水分子也可以起这种作用。丝氨酸、组氨酸和天冬氨酸的排列的侧链构成大多数丝氨酸蛋白酶中常见的催化三联体(catalytic triad)。丝氨酸蛋白酶或丝氨酸内肽酶组成一类肽酶,它们的特征是在酶的活性中心存在丝氨酸残基。丝氨酸蛋白酶参与机体的各种功能,包括凝血、炎症和原核和真核生物中的消化酶类。丝氨酸蛋白酶的活性位点呈裂缝形式,该裂缝是多肽底物结合的地方。氨基酸残基从多肽底物的N至C端(Pi,…,P3,P2,P1,P1′,P2′,P3′,…,Pj)以及它们各自的结合亚位点(Si,…,S3,S2,S1,S1′,S2′,S3′,…,Sj)标记。裂解在P1和P1’之间被催化。
如本文所用,“GH多肽的部分”是指展示全长多肽的一或多种生物学活性的任何部分。
如本文所用,“生长激素”是指由自然界生物产生的生长激素。例如,人产生垂体生长激素和胎盘生长激素。举例性的天然人垂体生长激素和胎盘生长激素见SEQ ID NO:1,712和713。其它动物如哺乳动物也产生天然生长激素,例如,牛天然生长激素例如SEQ ID NO:714,和猕猴天然生长激素例如SEQ ID NO:715。
如本文所用,“生长激素介导的疾病、病症或状况”是指用生长激素治疗能改善疾病或病症的任何症状或表现的任何疾病或病症。举例性的生长激素介导的疾病和病症包括但不限于生长缺陷疾病(如特纳氏综合征、子宫内生长迟缓、特发性身材矮小症、PraderWilli综合征或地中海贫血),AIDS消瘦、衰老、HIV感染对象受损的免疫功能、分解代谢疾病、手术后恢复、充血性心肌病、肝移植、肝切除术后肝再生、慢性肾功能衰竭、肾性骨营养不良、骨质疏松、软骨发育不全/季肋发育不全、骨骼发育异常、慢性炎性或营养性疾病(如克罗恩氏病)、短肠综合征、少年慢性关节炎、囊性纤维化、男性不育症、X连锁低磷血症性佝偻病、唐氏综合症、脊柱裂、努南综合征、肥胖症、肌肉强度受损以及纤维肌痛。
如本文所用,“生长激素缺陷”是指其中对象产生的天然生长激素的量比正常生长、发育和/或代谢所需的量低或不足的任何疾病、病症或状况。生长激素缺陷可以由高级脑下丘脑或垂体中的GH轴的破坏所致。这种功能障碍可以是先天的或获得性的。先天性生长激素缺陷可以与异常脑垂体(在MRI上可见)相关或可以是综合征的一部分,如视隔发育不良(de Morsier综合征),其可以包括其它的垂体缺陷、视神经发育不全和无透明隔。另一方面,获得性生长激素缺陷可以是由于创伤、感染(例如脑炎、脑膜炎)、颅辐射(亲躯体细胞看起来是垂体中对辐射最敏感的细胞)和/或其它全身性疾病(例如组织细胞增多病)所致。
如本文所用,“治疗”是指其中状况、病症或疾病的症状被改善或其它有利地改变的任何方式。治疗也包括本文提供的修饰的生长激素和组合物的任何药物学应用。
“治疗”患有疾病或状况的对象是指患者的症状被部分或全部减轻,或者在治疗后保持不变(static)。已被治疗的对象会显示症状的部分或全部减轻。因此治疗包括预防、治疗(therapy)和/或治愈(cure)。治疗也包括本文提供的修饰的生长激素和组合物的任何药物学应用。
本文所用短语“治疗有效量”或“治疗有效剂量”是指物质、化合物、材料或化合物的组合物的量是有效产生希望的治疗效果(例如疾状况状的改善)的量。
用本发明方法治疗的“患者”或“对象”可以是哺乳动物(即人或非人动物)。在一个实施方案中,哺乳动物可以是灵长类(例如人、黑猩猩、大猩猩、猴等)、家养动物(例如狗、马、猫、猪或奶牛)或啮齿类(例如小鼠或大鼠)等。特别地,哺乳动物是人。
如本文所用,“定向进化方法“是指“改造(adapt)”蛋白质、包括天然蛋白质、合成蛋白质或蛋白质结构域以使其具有改变的性质(proportions)(如在不同的或现存的天然或人工化学或生物学环境中起作用和/或引起新功能和/或增加或降低给定的活性和/或调节给定特征的能力)的方法。举例性的定向进化方法特别包括美国专利申请系列号10/022,249和美国公开申请US-2004-0132977-A1所述的合理定向进化方法。
如本文所用,“二维合理诱变扫描(2D扫描)”是指本文提供的方法,其中特定蛋白质序列的二维被扫描:(1)一维是鉴别蛋白质序列的氨基酸残基以用不同的氨基酸置换,被称为is-HIT靶位置,(2)第二维是选择用于置换特定的is-HIT靶的氨基酸类型,被称为置换氨基酸(replacing amino acid)。
如本文所用,“in silico”是指用计算机进行的研究和实验。Insilico方法包括但不限于分子建模研究和生物分子停靠实验。
如本文所用,“is-HIT”是指在靶蛋白质序列上in silico鉴别的氨基酸位置,其根据i)待被进化的特定蛋白质性质,ii)蛋白质的氨基酸序列和/或iii)各个氨基酸的已知性质而被鉴别。蛋白质序列上的这些is-HIT位置未经实验生物学方法而被鉴别。例如,一旦选定待优化的蛋白质特征,则研究各种信息源或先前知识(例如蛋白质的一级、二级或三级结构、文献、专利)以确定适于通过用不同的氨基酸置换而改善的蛋白质适合度(fitness)的氨基酸位置。这一步骤使用“in silico”蛋白质分析。沿靶蛋白质的一级序列的可能参与被进化的特征的所有可能的候选氨基酸位置在本文中被称为“in silicoHIT”(“is-HIT”)。在这一步骤中鉴别的所有is-HIT的文库(集合)代表了本文提供的二维扫描方法的第一维(靶残基位置)。
如本文所用,在鉴别is-HIT的上下文中提到的“适于提供进化的预定性质或活性”是指蛋白质上的这样的氨基酸位置,即,基于in silico分析,据信它们具有当被置换时导致所希望的被进化的活性的性质或特征。在鉴别置换氨基酸的上下文中提到的短语“适于提供进化的预定性质或活性”是指特定的氨基酸类型,基于in silico分析,据信其具有当被用于置换未修饰的起始蛋白质中的氨基酸时导致所希望的被进化的活性的性质或特征。
如本文所用,“高通量筛选(HTS)”是指测试大量样品如测试蛋白质样品或含有编码感兴趣蛋白质的核酸的细胞的样品以鉴别感兴趣的结构或鉴别与变体蛋白质或含有其的细胞相互作用的测试化合物的方法。HTS操作适于自动化并且典型地是计算机化的以进行样品制备、测定程序和大容量数据的随后处理。
如本文所用,术语“限制(restricted)”当用在鉴别选择用于氨基酸置换的蛋白质序列上的is-HIT氨基酸位置和/或鉴别置换氨基酸的上下文中时,是指蛋白质主链上的少于全部氨基酸被选择用于氨基酸置换;和/或少于其余的全部19个可利用的用于置换未修饰的起始蛋白质中的原始氨基酸的氨基酸被选择用于置换。在本发明方法的特定实施方案中,is-HIT氨基酸位置是受限制的,从而蛋白质主链上的少于全部氨基酸被选择用于氨基酸置换。在其它实施方案中,置换氨基酸是受限制的,从而少于其余的全部19个可利用的用于置换未修饰的起始蛋白质中的天然氨基酸的氨基酸被选择用于置换氨基酸。在一个举例性的实施方案中,鉴别is-HIT氨基酸位置和置换氨基酸的扫描均是受限制的,从而蛋白质主链上的少于全部氨基酸被选择用于氨基酸置换并且少于其余全部19个可利用的用于置换天然氨基酸的氨基酸被选择用于置换。
如本文所用,“候选LEAD”是突变蛋白质,据信它们潜在具有在任何希望有改变的属性、化学、物理或生物学性质中的改变。在本文的方法中,候选LEAD通常如下产生,即典型地用如使用PAM分析获得的其余19个氨基酸的限制亚集或全部来系统性置换蛋白质或其结构域中的is-HIT位置。候选LEAD可以通过用本文的高通量方法测试的本领域技术人员已知的其它方法产生。
如本文所用,“LEAD”是已证实其活性针对特定的属性、化学、物理或生物学性质被优化或改善的“候选LEAD”。本文中,针对感兴趣的功能,“LEAD”典型地具有与未修饰的和/或野生型(天然)蛋白质相比至少10%、20%、30%、40%、50%、60%、70%、80%、90%、100%、150%、200%或更高的不同的活性。在某些实施方案中,活性的改变是未修饰的靶蛋白质的活性的至少约5%、10%、15%、20%、25%、30%、35%、40%、45%、50%、55%、60%、65%、70%、75%、80%、85%、90%、95%或100%。在其它实施方案中,活性的改变不超过未修饰的靶蛋白质的活性的约5%、10%、15%、20%、25%、30%、35%、40%、45%、50%、55%、60%、65%、70%、75%、80%、85%、90%、95%或100%。在另外的实施方案中,活性的改变是比未修饰的靶蛋白质的活性高至少约2倍、3倍、4倍、5倍、6倍、7倍、8倍、9倍、10倍、20倍、30倍、40倍、50倍、60倍、70倍、80倍、90倍、100倍、200倍、300倍、400倍、500倍、600倍、700倍、800倍、900倍、1000倍或更多倍。希望的改变可以是活性的增加或降低,其取决于感兴趣的功能或性质(例如~10%,~20%,等)。LEAD可以被进一步优化,即置换同一蛋白质分子上的多个(2个或多个)″is-HIT″靶位置以产生“super-LEAD”。
如本文所用,术语“super-LEAD”是指通过将存在于两个或多个LEAD分子中的单突变组合成一个单蛋白质分子而获得的蛋白质突变体(变体)。因此,在本文提供的修饰的蛋白质的上下文中,短语“包含一或多个单氨基酸置换的蛋白质”包括两个或多个本文针对各蛋白质描述的突变的任何组合。例如,本文提供的具有一或多个单氨基酸置换的修饰的蛋白质可以具有在所揭示的置换位置的2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20或更多个氨基酸置换的任何组合。产生super-LEAD突变体分子的集合,测试并在可寻址阵列中逐一进行表型鉴定。Super-LEAD突变体分子含有变化数目和类型的LEAD突变。展示对被进化的特定特征的进一步改善的适合度的分子被称为super-LEAD。Super-LEAD可以通过本领域技术人员已知的其它分子产生并且通过本文描述的高通量方法测试。针对感兴趣的功能,super-LEAD典型地具有以所希望量不同于LEAD的改善的活性的活性,例如不同于其所衍生的至少一种LEAD突变体至少10%、20%、30%、40%、50%、60%、70%、80%、90%、100%、150%、200%或更高。与LEAD一样,super-LEAD活性的改变取决于被“进化”的活性。希望的改变可以是活性增加或降低,其取决于感兴趣的功能或性质。
如本文所用,描述修饰的生长激素与未修饰的生长激素相比具有高于抗炎活性(或另一种活性)的增殖活性(或其它活性)是比较每种与未修饰的或天然形式相比的活性的变化绝对值。
如本文所用,短语“改变的位置(altered loci)”是指LEAD或super-LEAD中用不同的置换氨基酸置换导致产生希望的被改变的表型或活性的is-HIT氨基酸位置。
如本文所用,“暴露的残基”使其超过15%的表面暴露于溶剂。
如本文所用,短语“结构同源性”是指两个或多个蛋白质主链之间的空间重合程度。采取相同的蛋白质结构、折叠并显示三维结构空间重叠(superposition in space)相似性的蛋白质主链可以被认为是结构同源的。结构同源性不是基于序列同源性而是基于三维同源性。基于蛋白质之间的结构同源性而被认为是同源的两个不同蛋白质中的两个氨基酸不是必须位于基于序列的同源区内。例如,在给定的空间位置或互相之间限定的区域具有小于3.5、3.0、2.5、2.0、1.7或1.5埃(_)的均方根(RMS)偏差的蛋白质主链可以被认为是在该区域结构同源的,并被称为在它们的主链之间具有“高重合”。本发明也包括,位于具有某种程度结构同源性的两或多个不同蛋白质序列上的基本上等价的(例如“结构相关的”)氨基酸位置具有相当的功能任务,并且在本文中也被称为“结构同源位置”。这两个氨基酸随后可被称为彼此结构相似或结构相关,即使它们在氨基酸序列上的精确一级线性位置当进行序列对比时彼此不匹配。当这些序列根据经典序列同源性规则进行序列对比时,结构相关的氨基酸在一级氨基酸序列中可以彼此相距甚远。如本文所用,“结构同系物”是通过结构同源性产生的蛋白质。
如本文所用,“未修饰的靶多肽”、“未修饰的多肽”、“未修饰的细胞因子”、“未修饰的GH”、“未修饰的生长激素”或其语法学变异名称是指选择用于使用本文提供的方法进行修饰的起始多肽(蛋白质)。起始未修饰的靶多肽可以是天然发生的野生型蛋白质。另外,起始未修饰的靶多肽先前可以已经被改变或突变,从而它们与天然野生型同种型不同,但是相对于在本文产生的随后修饰的多肽,也在本文中被称为起始未修饰的靶多肽。因此,先前已被修饰以与未修饰的参考蛋白质相比具有特定生物学活性的希望的增加或降低的现有技术已知的现有多肽可以被选择并在本文中用作起始“未修饰的靶蛋白质”。例如,已通过一或多个单氨基酸改变而从其天然形式修饰而来并且具有所希望活性如对蛋白酶解的抗性的增加或降低的多肽可以用于本发明方法中作为起始未修饰的靶多肽以用于对相同或不同生物学活性的进一步修饰。
类似地,先前已被修饰以与未修饰的或参考蛋白质相比具有特定生物学活性的希望的改变如增加或降低的现有技术已知的现有多肽可以被选择并用在本发明中以鉴别其它结构同源的靶多肽上的结构同源位置。例如,已通过一或多个单氨基酸改变而修饰并且具有所希望活性(如对蛋白酶解的抗性)的增加或降低的多肽可以用于本发明方法中以鉴别结构同源的靶多肽上的可以被合适的置换氨基酸置换的相应的结构同源位置并测试所希望生物学活性的增加或降低。
如本文所用,“变体”或“生长激素变体”或“修饰的生长激素”是指与未修饰的GH多肽相比具有一或多个突变的生长激素多肽。所述一或多个突变可以是一或多个氨基酸置换、插入、缺失和/或其任意组合。
如本文所用,“相应于蛋白质的氨基酸位置的一个或多个位置”是指基于与一规定的参考蛋白的序列和/或结构对比而确定彼此相应的氨基酸位置。例如,相应于SEQ ID NO:1的人生长激素的一个氨基酸位置的位置可以通过将SEQ ID NO:1的氨基酸序列与一感兴趣的特定GH多肽进行对比而经验性确定。相应的位置可以通过本领域技术人员使用手工对比或使用各种可利用的对比程序(例如,BLASTP)进行这类对比而确定。相应的位置也可以基于结构对比,例如,使用计算机模拟的蛋白质结构的对比。描述一个多肽的氨基酸相应于一公开序列中的氨基酸是指使用标准对比算法如GAP算法将所述多肽与所述公开序列进行对比以最大化相同性或同源性(当对比保守氨基酸时)时鉴别的氨基酸。如本文所用,“在相应于...的位置”是指核酸分子或蛋白质中的相对于另一参考核酸分子或蛋白质中的位置的感兴趣的位置(即碱基编号或残基编号)。相应于另一参考蛋白质中的位置的感兴趣位置可以在例如前体蛋白质、等位基因变体、异源蛋白质、来自另一物种的相同蛋白质的氨基酸序列等中。相应的位置可以通过比较和对比序列以最大化匹配的核苷酸或残基数而确定。例如,序列之间的相同性可以大于95%、大于97%、大于98%,更特别地高于99%。感兴趣的位置然后被给予在参考核酸分子或多肽序列中被赋予的编号。本领域普通技术人员应理解对于与成熟的未修饰GH多肽相比的修饰的GH多肽,修饰的多肽的氨基酸残基1相应于成熟的未修饰GH多肽的氨基酸残基1。本领域普通技术人员还应理解对于与前体未修饰GH多肽相比的修饰的前体GH多肽,修饰的多肽的氨基酸残基1相应于未修饰的GH多肽的氨基酸残基1。另外,修饰的前体GH多肽的氨基酸残基27相应于成熟的未修饰的GH多肽的氨基酸残基1,这是由于存在前体多肽的26个氨基酸长的信号序列。因此,本领域普通技术人员能够确定修饰的和未修饰的GH多肽中的“相应的”氨基酸残基。
如本文所用,术语“同源性”和“相同性”是可互换使用的,但是蛋白质的同源性可以包括保守氨基酸改变。通常,为了鉴别相应的位置,氨基酸序列被对比以便获得最高级匹配(参见,例如,:Computational Molecular Biology,Lesk,A.M.,ed.,Oxford UniversityPress,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.,HumanaPress,New Jersey,1994;
Sequence Analysis in Molecular Biology,vonHeinje,G.,Academic Press,1987;and
Sequence Analysis Primer,Gribskov,M.and Devereux,J.,eds.,M Stockton Press,New York,1991;Carillo et al.SIAM J.Applied Math 48:1073(1988))。
如本文所用,“序列相同性”是指相同的氨基酸的数目(同源性还包括保守氨基酸取代)。序列相同性可以通过标准的对比算法程序使用由每一供应商建立的默认缺口罚分来确定。实质上同源的核酸分子典型地在中等严格性或高严格性下沿核酸全长或沿感兴趣的全长核酸分子的至少约70%、80%或90%杂交。本发明还包括含有替换杂交核酸分子中的密码子的简并密码子的核酸分子。)对于蛋白质,为了确定同源性,保守氨基酸可以作为相同氨基酸被对比,在这种情况下,相同性百分比和同源性百分比是不同的)。任何两个核酸分子是否具有至少80%、85%、90%、95%、96%、97%、98%或99%“相同的”核苷酸序列可以用已知的计算机算法如″FAST A“程序”使用例如Pearson et al.Proc.Natl.Acad.Sci.USA 85:2444(1988)中所述的默认参数而确定(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.,J.Molec.Biol.215:403(1990);Guide to Huge Computers,Martin J.Bishop,ed.,Academic Press,SanDiego,1994,and Carillo et al.SIAM J.Applied Math 48:1073(1988))。例如,国立生物技术信息中心数据库的BLAST函数可以被用于确定相同性。其它可商购或公共程序包括DNAStar″”MegAlign”程序(Madison,WI)和University of Wisconsin Genetics Computer Group(UWG)“Gap”程序(Madison WI))。蛋白质和/或核酸分子的同源性或相同性百分比可以例如通过用经Smith and Waterman(Adv.Appl.Math.2:482(1981))修改的GAP计算机程序(例如Needleman et al.J.Mol.Biol.48:443(1970)比较序列信息而确定。简而言之,GAP程序对相似性的定义为相似的被对比符号(即核苷酸或氨基酸)数目除以两个序列中较短者的总符号数目。GAP程序的默认参数可以包括:(1)一元比较矩阵(值1为相同性,0为非相同性)和Gribskov et al.Nucl.Acids Res.14:6745(1986)的权重比较矩阵(weighted comparisonmatrix),如Schwartz and Dayhoff,eds.,
ATLAS OF PROTEIN SEQUENCE AND STRUCTURE,National Biomedical ResearchFoundation,pp.353-358(1979)所述;(2)每一缺口罚分3.0,每一缺口中的每一符号额外罚分0.10;以及(3)末端缺口无罚分。因此,如本文所用,术语“相同性”代表测试和参考多肽或多核苷酸之间的比较。
如本文所用,术语“与...至少90%相同”是指相对于参考多肽的相同性百分比为90%至100%。90%或更高水平的相同性代表如下事实,假设用于举例目的比较长度为100个氨基酸的测试和参考多肽,测试多肽中不超过10%(即100个中有10个)的氨基酸与参考多肽中的不同。在测试和参考多核苷酸之间可以进行类似的比较。这种差异可以以随机分布于整个长度的氨基酸序列中的点突变为代表,或者它们可以聚集在变化长度的至多为所允许的最大值例如10/100氨基酸差异(约90%相同性)的一或多个位置中。差异可以被定义为核酸或氨基酸取代、插入或缺失。在高于约85-90%的同源性或相同性水平,结果应该独立于程序和缺口参数集,这种高相同性水平可以容易地评估,经常不依赖于软件。
如本文所用,“单氨基酸置换”是指一个氨基酸由另一个氨基酸置换。置换可以是由天然氨基酸或非天然氨基酸置换。当蛋白质中的一个氨基酸被另一个氨基酸置换时,该蛋白质中的氨基酸总数不变。
如本文所用,术语“在每一靶蛋白质上仅发生一个氨基酸置换”是指靶蛋白质被修饰从而其与未修饰形式的靶蛋白质仅有一个单氨基酸改变的不同。例如,在一个实施方案中,通过在蛋白质主链上的仅一个is-HIT靶位置的单氨基酸残基的置换而进行诱变(例如在可寻址阵列上“一个接一个(″one-by-one″)进行”,从而所产生的每个突变体是每个单诱变反应的单一产物。单氨基酸置换诱变反应在每一is-HIT靶位置针对所选择的每一置换氨基酸重复进行。由此,产生多个突变蛋白质分子,其中每个突变蛋白质在仅一个is-HIT靶位置含有单氨基酸置换。
如本文所用,术语“假野生型”在指单或多氨基酸置换时,是这样的氨基酸,它们尽管与一给定氨基酸位置上的原始如天然氨基酸不同,但是可以置换该位置的天然氨基酸而不会导致一特定蛋白质活性的任何可测得的改变。产生编码突变分子集合的核酸分子集合群并进行表型鉴定,以便选择具有不同于原始氨基酸的氨基酸序列但是仍能引起与原始蛋白质基本上相同水平(即至少10%、50%、70%、90%、95%、100%,取决于蛋白质)和类型的希望活性的蛋白质。
如本文所用,“裸多肽链”是指未经翻译后修饰或其它化学修饰并且仅含共价连接的氨基酸的多肽。
如本文所用,“多肽复合物”包括通过化学修饰或翻译后修饰产生的多肽。这些修饰包括但不限于聚乙二醇化、白蛋白化(albumination)、糖基化、法呢基化(farnysylation)、磷酸化和/或本领域已知的其它多肽修饰。
如本文所用,“输出信号”是指可以随时间跟踪及如果需要进行定量的参数。例如,当重组蛋白被导入细胞中时,含有重组蛋白的细胞经历一些变化。可以被监测并用于评价转化或转染的任何这类变化就是输出信号,而所述细胞被称为报道细胞;编码核酸称为报道基因,包括编码核酸的构建体是报道构建体。输出信号包括但不限于酶活性、荧光、发光、产生的产物的量和其它这类信号。输出信号包括基因或基因产物、包括插入到质粒病毒中的异源基因(转基因)的表达。输出信号是时间(“t”)的函数并与组合物中使用的蛋白质的量相关。对于高浓度的蛋白质,输出信号可以高或低。对于任何特定浓度,输出信号作为时间的函数而增加直至达到平台。输出信号也可以测量表达异源基因的细胞与生物学制剂之间的相互作用。
如本文所用,Hill方程是将药物(即测试化合物或物质)浓度与所测的应答联系起来的数学模型
其中y是测量的变量,如应答、信号,ymax是可实现的最大应答,[D]是药物的摩尔浓度,[D50]是产生对药物的50%最大应答的浓度,n是斜率参数,如果药物结合单位点并且位点之间或之中无协同性则n是1。Hill图是配体占据的受体与游离受体之比的log10vs.log[D](M)。斜率是n,其中斜率大于1表示结合位点中的协同性,斜率小于1可以表示结合的异质性。这一方程已被用于评价复杂生物学系统中的相互作用的方法中(参见基于PCT No.PCT/FR00/03503公开的国际PCT申请WO 01/44809)。
如本文所用,在基于Hill的分析中(基于PCT No.PCT/FR00/03503公开的国际PCT申请WO 01/44809),参数π,κ,τ,ε,η,θ如下:
π 是作用于测定(基于细胞的)系统上的生物制剂的效力;
κ 是测定系统引起对生物制剂的应答的抗性常数;
ε 生物制剂在测定系统上触发的过程或反应的整体效率;
τ 是生物制剂的表观效价;
θ 是生物制剂的绝对效价;以及
η 是生物学过程或反应的异质性。
特别地,如本文所用,参数π(效力)或κ(抗性常数)分别用于评价测试制剂在测定系统中产生应答的效力和测试系统对应答所述制剂的抗性。
如本文所用,ε(效率)是在Hill曲线(或者通常任何其它S形或线性逼近)的拐点的斜率,用于估算引起生物学或药理学应答的整体反应(生物制剂和测定系统一起)的效率。
如本文所用,τ(表观效价)用于测量生物制剂的有限稀释或表观效价。
如本文所用,θ(绝对效价)用于测量生物制剂的绝对有限稀释或效价。
如本文所用,η(异质性)测量的是整体反应中存在的非连续相,其反映在Hill系数值或抗性常数中的陡变。
如本文所用,“编码突变体文库(集合)的核酸分子集合群(population of sets)”是指携带(编码)基因变体的质粒或其它载体的文库,其中个体质粒或其它个体载体携带个体基因变体。文库的每一元素(成员)互相物理分离,如一个一个单独地在合适的可寻址阵列中,并且已作为独立诱变反应的单一产物而产生。当提及这种蛋白质文库时,就如此称谓。
如本文所用,“报道细胞”是“报道”即经历应答一种条件而产生的变化的细胞,所述条件例如是暴露于一种蛋白质或病毒或者改变其外部或内部环境。
如本文所用,“报道分子”或“报道部分”是指使得能检测感兴趣分子如由细胞表达的蛋白质的任何部分。报道部分包括但不限于,例如,荧光蛋白如红、蓝、绿荧光蛋白、LacZ和其它可检测蛋白和基因产物。为在细胞中表达,编码报道部分的核酸分子可以作为与感兴趣蛋白质的融合蛋白而表达或者在感兴趣的启动子的控制下表达。
如本文所用,“表型”是指基因型(基因序列)的物理、生理或其它表现。在本文的方法中,评估由基因型的改变导致的表型。
如本文所用,“活性”在该术语的最广义中是指在一个系统(生物学、化学或物理系统)中由蛋白质或蛋白质突变体在与系统相互作用时引起的任何性质的任何变化(酶反应中产物量的变化、在细胞增殖、免疫原性、毒性中的变化)。另外,本文中描述对蛋白酶的抗性、蛋白酶解、与血清或血液保温时使用术语“活性”、“更高的活性”或“更低的活性”是指在蛋白酶/血液或血清处理“之后”与蛋白酶/血液或血清处理“之前”之间的比率或残留生物学活性(细胞增殖)。
如本文所用,“活性”是指待被进化的功能或性质。活性位点是指负责或参与赋予活性或功能的位点。被进化的活性或活性位点(功能或性质以及赋予或参与赋予活性的位点)可以与蛋白质的天然活性无关。例如,其可以是蛋白质上的赋予免疫原性的“活性位点”(免疫原性位点或表位)。
如本文所用,“活性”、“生物学活性”和“药理学活性”生物学药剂的任何活性,包括但不限于对蛋白酶解的抗性、生物学效率、转导效率、基因/转基因表达、差异基因表达和诱导活性、效价、后代生产力、毒性、细胞毒性、免疫原性、细胞增殖和/或分化活性、形态发生活性、治疗活性、增加的肌肉质量、药理学活性、细胞/组织向性和输送。如本文所用,修饰的GH多肽保留至少1%、2%、3%、4%、5%、6%、7%、8%、9%、10%、...20%、...30%、...40%、...50%、...60%、...70%....80%、...90%、...95%、96%、97%、98%或至少99%的未修饰的GH多肽的活性。在其它的实施方案中,活性的变化是比未修饰的GH高至少约2倍、3倍、4倍、5倍、6倍、7倍、8倍、9倍、10倍、20倍、30倍、40倍、50倍、60倍、70倍、80倍、90倍、100倍、200倍、300倍、400倍、500倍、600倍、700倍、800倍、900倍、1000倍或更多倍。活性可以例如使用下述实施例中描述的那些测定进行测量。
如本文所用,当氨基酸出现在本文提供的各种氨基酸序列中时是根据它们的已知三字母或单字母缩写进行描述(见表1)。当核苷酸出现在各种核酸片段中时是用本领域常规使用的标准单字母名称表示。
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。多肽包含两个或更多个氨基酸。本文中,氨基酸包括20个天然发生的氨基酸、非天然氨基酸和氨基酸类似物。它们包括其中α-碳具有侧链的氨基酸。
如本文所用,除非特别指明,否则任何保护基、氨基酸和其它化合物的缩写是根据它们的通用名、公认的缩写或IUPAC-IUBCommission on Biochemical Nomenclature,Biochem.11:1726(1972))。每个天然发生的L-氨基酸由标准三字母代码(或单字母代码)或者带有前缀“L-”的标准三字母代码(或单字母代码)表示,前缀“D-”表示氨基酸的立体异构形式是D。
如本文所用,氨基酸残基是指在多肽的肽连键处化学消化(水解)时形成的氨基酸。本文描述的氨基酸残基假定为“L”异构形式。“D”异构形式的残基可以取代任何L-氨基酸残基,只要多肽保留所希望的功能性质即可。NH2是指多肽的氨基末端的游离氨基基团。COOH是指存在于多肽的羧基末端的游离羧基。为与J.Biol.Chem.,243:3552-3559(1969)描述的并被37 C.F.R.§§1.821-1.822采用的标准多肽命名一致,氨基酸残基的缩写如表1所示:
表1
对应表
符号 | ||
单字母 | 三字母 | 氨基酸 |
Y | Tyr | 酪氨酸 |
G | Gly | 甘氨酸 |
F | Phe | 苯丙氨酸 |
M | Met | 甲硫氨酸 |
A | Ala | 丙氨酸 |
S | Ser | 丝氨酸 |
I | Ile | 异亮氨酸 |
L | Leu | 亮氨酸 |
T | Thr | 苏氨酸 |
V | Val | 缬氨酸 |
P | Pro | 脯氨酸 |
K | Lys | 赖氨酸 |
H | His | 组氨酸 |
Q | Gln | 谷氨酰胺 |
E | Glu | 谷氨酸 |
Z | Glx | Glu和/或Gln |
W | Trp | 色氨酸 |
R | Arg | 精氨酸 |
D | Asp | 天冬氨酸 |
N | Asn | 天冬酰胺 |
B | Asx | Asn和/或Asp |
C | Cys | 半胱氨酸 |
X | Xaa | 未知或其它 |
应注意本文中由分子式表示的所有氨基酸残基序列均是从左至右为传统的氨基末端至羧基末端的方向。另外,术语“氨基酸残基”广义上包括对应表(表1)所列的氨基酸以及修饰的和不常见的氨基酸,如并入本文作参考的37 C.F.R.§§1.821-1.822所述的那些。另外,应注意氨基酸残基序列开始和结尾处的短破折号(-)代表与其它的一或多个氨基酸残基的序列或与氨基端基如NH2或与羧基端基如COOH的肽键。
如本文所用,“天然发生的”氨基酸是指存在于多肽中的20个L-氨基酸。
如本文所用,术语“非天然氨基酸”是指具有类似于天然氨基酸的结构但被结构修饰以模拟天然氨基酸的结构和反应性的有机化合物。非天然发生的氨基酸因此包括除20个天然发生的氨基酸之外的氨基酸或氨基酸类似物,并且包括但不限于氨基酸的D-立体异构体。举例性的非天然氨基酸在本文中有描述并且是本领域技术人员已知的。
如本文所用,核酸包括DNA、RNA及其类似物,包括蛋白核酸(PNA)及其混合物。核酸可以是单链或双链的。当描述可以任选地用可检测标记如荧光或放射性标记来标记的探针或引物时,单链分子在包括在内。这类分子典型的长度是它们统计学独特地具有低拷贝数(典型地小于5个,通常小于3个)用于探查或引导一个文库。通常探针或引物含有与感兴趣基因互补或相同的至少14、16或30个连续核苷酸的序列。探针和引物长度可以是10、14、16、20、30、50、100或更多个核苷酸碱基。
如本文所用,两个或多个蛋白质如GH蛋白和其它细胞因子上的“相应的结构相关的”位置是指基于使蛋白质之间的三维重叠最大化的结构同源性确定的氨基酸位置。
如本文所用,术语“相同性”代表测试和参考多肽或多核苷酸之间的比较。例如,测试多肽可以被定义为与参考多肽90%或更高相同的任何多肽。如本文所用,术语至少“90%相同性”是指相对于参考多肽从90%至100%的相同性百分比。90%或更高水平的相同性代表如下事实,假设用于举例目的比较长度为100个氨基酸的测试和参考多肽,测试多肽中不超过10%(即100个中有10个)的氨基酸与参考多肽中的不同。在测试和参考多核苷酸之间可以进行类似的比较。这种差异可以以随机分布于整个长度的氨基酸序列中的点突变为代表,或者它们可以聚集在变化长度的至多为所允许的最大值例如10/100氨基酸差异(约90%相同性)的一或多个位置中。差异可以被定义为核酸或氨基酸取代、插入或缺失。
如本文所用,术语“序列相关的蛋白质”是指彼此具有至少50%、至少60%、至少70%、至少80%、至少90%或至少95%氨基酸相同性或同源性。
如本文所用,不相关蛋白质或“序列不相关蛋白质”家族是指彼此具有小于50%、小于40%、小于30%或小于20%氨基酸相同性或同源性的蛋白质。
如本文所用,还应理解术语“基本上相同”或“相似”随着上下文而不同,这为相关领域技术人员所理解。
如本文所用,异源或外来核酸如DNA和RNA可互换使用,是指不是作为其所存在的基因组的一部分而天然存在的DNA或RNA,或者是发现在基因组中的与其天然存在的基因座不同的基因座中的DNA或RNA。异源核酸包括对于其被导入的细胞是非内源的并且得自另一细胞或合成制备的核酸。通常,尽管不是必须,这种核酸编码不是表达其的细胞正常产生的RNA和蛋白质。本文中异源DNA包含本领域技术人员所认知或认为是针对表达其的细胞或基因座是异源或外来的任何DNA或RNA。异源DNA和RNA也可以编码通过影响转录、翻译或其它可调控生物化学过程而介导或改变内源性DNA的表达的RNA或蛋白质。异源核酸的例子包括但不限于编码可示踪标记蛋白质如赋予药物抗性的蛋白质的核酸,编码药物学有效的物质如抗癌剂、酶和激素的核酸,以及编码其它类型的蛋白质如抗体的DNA。
因此,本文中“异源DNA”或“外来DNA”包括不以基因组中发现的对应DNA分子的精确方向和位置存在的DNA分子。其也可以指来自另一生物体或物种的DNA分子(即外源的)。
如本文所用,“分离的核酸分子或多肽或其它生物分子”是指核酸或多肽已经与获得所述多肽或核酸的遗传环境脱离。其也可以指从天然状态改变的。例如,如本文采用的该术语,天然存在于活体动物中的多核苷酸或多肽不是“分离的”,但是与其天然状态共存的材料分离的相同多核苷酸或多肽是“分离的”。因此,重组宿主细胞产生的和/或含在其中的多肽或多核苷酸被认为是分离的。“分离的多肽”或“分离的多核苷酸”还包括已从重组宿主细胞或从天然来源部分纯化或基本上纯化的多肽或多核苷酸。例如,重组产生的化合物可以用Smith et al.,Gene,67:31-40(1988)所述的一步法基本上纯化。术语分离的和纯化的有时候互换使用。
因此,“分离的”是指核酸不含位于生物体的天然发生的基因组(如果有)中编码感兴趣核酸的基因的紧邻侧翼的那些基因的编码序列。分离的DNA可以是单链或双链的,并且可以是基因组DNA、cDNA、重组杂交DNA或合成DNA。其可以与起始DNA序列相同,或者可以由于一或多个核苷酸的缺失、添加或取代而与这种序列不同。
当“分离的”或“纯化的”是指从生物细胞或宿主中制备的制备物时,其意味着含有所指示DNA或多肽的任何细胞提取物,包括感兴趣DNA或多肽的粗提取物。在多肽的情况中,例如,纯化的制备物可以通过个别技术或一系列制备或生物化学技术而获得,并且感兴趣的DNA或多肽可以以各种纯度存在于这些制备物中。这些技术可以包括例如但不限于硫酸铵分级分离、凝胶过滤、离子交换层析、亲和层析、密度梯度离心和电泳。
应理解“基本上纯的(substantially pure)”或“分离的”DNA或蛋白质制备物是指不含这种DNA或蛋白质在自然界中正常结合的天然发生的材料的制备物。“实质上纯的(Essentially pure)”应理解是指“高度”纯化的制备物,其含有至少95%的感兴趣DNA或蛋白质。
应理解含有感兴趣DNA或蛋白质的细胞提取物是指得自表达感兴趣蛋白质或含有感兴趣DNA的细胞的均质制备物或无细胞制备物。术语“细胞提取物”包括细胞培养基,特别是已除去细胞的用过的培养基。
如本文所用,“靶向剂”是指可以结合另一个靶分子的任何分子,如抗体、受体或配体。
如本文所用,“受体”是指特异性结合至其它分子(或与之结合)的生物活性分子。术语“受体蛋白”可以更具体地用于指明特异受体的蛋白质性质。
如本文所用,“重组体”是指由基因工程产生的任何后代。
如本文所用,“启动子区”是指基因DNA的控制与其可操纵连接的DNA的转录的部分。启动子区包括足以用于RNA聚合酶识别、结合和转录起始的特异DNA序列。启动子区的这一部分称为“启动子”。另外,启动子区包括调节这一RNA聚合酶的识别、结合和转录起始活性的序列。启动子根据调节的性质可以是组成型的或由顺式或反式作用因子调节的。
如本文所用,术语“可操纵地连接”通常是指序列或节段已被共价连接进一段DNA,可以是单链或双链形式,其中在一个节段上的控制或调节序列控制或允许其它节段的表达或复制或其它这类控制。两个节段不是必须连续的。对于基因表达,DNA序列与调节序列如此方式连接以便当合适的分子例如转录激活蛋白与所述调节序列结合时能控制或允许基因表达。
如本文所用,用重组DNA方法经重组手段产生是指使用熟知的分子生物学方法表达由克隆的DNA编码的蛋白质,包括基因的克隆表达和方法,如基因改组和噬菌体展示以筛选希望的特异性。
如本文所用,“剪接变体”是指由基因组DNA的初级转录物的差异加工产生的变体,其导致超过一种类型的mRNA。
如本文所用,“组合物”是指两种或多种产物或化合物的任何混合物。其可以是溶液、悬浮液、液体、粉末、糊状物、水性、非水性,或其任意组合。
如本文所用,“组合”是指两或多个项目之间的任何结合。
如本文所用,“制品”是制造并出售的产品。本申请中,该术语包括含在包装物品中的修饰的生长激素多肽和核酸。
如本文所用,“试剂盒”或“药盒”是指本文提供的修饰的生长激素多肽或核酸分子与用于包括但不限于给药、诊断和生物学活性或性质的评价目的的另一项目的组合。
如本文所用,与一个产物“基本上相同”是指足够相似从而感兴趣的性质足够地未改变以便基本上相同的产物可以用于替换该产物。
如本文所用,术语“载体”是指能转运与其连接的另一个核酸的核酸分子。一种类型的举例性载体是附加体,即能够染色体外复制的核酸。举例性的载体是能够自主复制和/或表达与其连接的核酸的载体,这类载体典型地包括复制起点。也可以设计用于整合进宿主染色体中的载体。能够指导与其可操纵连接的基因的表达的载体在本文中称作“表达载体”。表达载体通常是“质粒”形式,其通常指环状双链DNA环,它们在载体形式下不与染色体结合。“质粒”和“载体”可互换使用,因为质粒是最常用形式的载体。
如本文所用,载体也包括“病毒载体”。病毒载体是工程病毒,其可操纵地与外源基因连接以(作为运载体或穿梭体)将外源基因转移进细胞。
如本文所用,“等位基因”与“等位基因变体”互换使用,是指基因或其部分的可变形式。等位基因在同源染色体上占据相同的基因座或位置。当对象具有一个基因的两个相同等位基因时,该对象被称为对该基因或等位基因是纯合的。当对象具有一个基因的两个不同等位基因时,该对象被称为对该基因是杂合的。一特异基因的等位基因彼此之间可以是单核苷酸或几个核苷酸的不同,并且可以包括核苷酸的取代、缺失和插入。一个基因的等位基因也可以是含有突变的基因形式。
如本文所用,术语“基因”或“重组基因”是指包含一个可读框并包括至少一个外显子和任选地内含子编码序列的核酸分子。基因可以是RNA或DNA。基因可以包括编码区之前和之后的区域(前导区和非转录尾区)。如本文所用,“内含子”是指在一给定基因中存在的在mRNA成熟期间被剪接除去的DNA序列。如本文所用,术语“编码序列”是指编码蛋白质中存在的氨基酸序列的基因部分。
如本文所用,“与编码SEQ ID NO:的氨基酸序列的核苷酸序列互补的核苷酸序列”是指编码具有特定SEQ ID NO:的氨基酸序列的核酸链的互补链的核苷酸序列。术语“互补链”在本文中与“互补序列”互换使用。一个核酸链的互补序列可以是编码链的互补序列或非编码链的互补序列。当指双链核酸时,编码具有特定SEQ IDNO:的氨基酸序列的核酸的互补序列是指编码特定SEQ ID NO:的氨基酸序列的链的互补链或者指具有该特定核酸序列的互补链的核苷酸序列的任何核酸。当指具有一核苷酸序列的单链核酸时,这一核酸的互补序列是具有与该特定核酸序列的核苷酸序列互补的核苷酸序列的核酸。
如本文所用,术语“有义链”是指双链核酸分子的具有mRNA序列的链,所述mRNA编码由该双链核酸分子编码的氨基酸序列。
如本文所用,术语“反义链”是指双链核酸分子的是mRNA序列的互补序列的链,所述mRNA编码由该双链核酸分子编码的氨基酸序列。
如本文所用,“阵列”是指含有三个或更多个成员的元件如核酸分子的集合。可寻址阵列是其中阵列成员可典型地由固相基质上的位置或由可鉴别或可检测标记如由颜色、荧光、电信号(即RF、微波或其它基本上不改变感兴趣分子的相互作用的频率)、条形码或其它符号学、化学或其它这类标记而鉴别的阵列。在某些实施方案中,阵列成员是固定化的,以将可鉴别的基因座分立在固相表面上或者直接或间接连接于或以其它方式结合于可鉴别标记如固定于(affixed to)微球或其它颗粒支持物(本文中是指珠)和悬浮于溶液或扩散在表面上。
如本文所用,“支持物”(也称为基质支持物、基质、可溶支持物或固体支持物)是指感兴趣分子、典型地为生物分子、有机分子或生物特异性配体所连接或接触的任何固体或半固体或不溶支持物。这些材料包括用作亲和基质或支持物以进行化学和生物分子合成和分析的任何材料,例如但不限于:聚苯乙烯、聚碳酸酯、聚丙烯、尼龙、玻璃、葡聚糖、壳多糖、沙、浮石、琼脂糖、多糖、树状物、buckyballs、聚丙烯酰胺、硅、橡胶以及其它用作固相合成、亲和分离和纯合、杂交反应、免疫测定和其它这类应用的支持物的材料。本文中基质可以是颗粒状的或可以是连续表面形式,如微滴板或孔、玻片、硅芯片、硝酸纤维素片、尼龙筛或其它这类材料。当呈颗粒状时,典型地颗粒有至少一维尺寸为5-10mm范围或更小。这种颗粒在本文中统称为“珠”,它们通常是但不是必须是球形的。然而这种描述不限制基质的几何形状,其可以是任何形状,包括随机形状、针、纤维和长形。大致为球形的珠、特别是可用于液相的微球也包括在内。珠可以包括另外的成分,如用于使用磁铁进行分离的磁性或顺磁性颗粒(参见例如Dynabeads(Dynal,Oslo,Norway)),只要所述另外的成分不干扰本文的方法和分析即可。
如本文所用,“基质”或“支持颗粒”是指分离颗粒形式的基质材料。颗粒具有任何性状和尺寸,但典型地有至少一维是100mm或更小、50mm或更小、10mm或更小、1mm或更小、100μm或更小、50μm或更小并且典型地具有100mm3或更小、50mm3或更小、10mm3或更小和1mm3或更小、100μm3或更小的尺寸并且可以是立方微米级。这些颗粒统称“珠”。
如本文所用,除非特别指明,否则任何保护基、氨基酸和其它化合物的缩写是根据它们的通用名、公认的缩写或IUPAC-IUBCommission on Biochemical Nomenclature,Biochem.11:1726(1972))。
B.生长激素
生长激素(GH)也已知为促生长素(somatotropin),是一种在包括哺乳动物在内的动物中产生的肽激素。GH是由垂体前叶的促生长激素细胞产生的细胞因子蛋白质家族的成员。GH通过对蛋白质、碳水化合物和脂质代谢的作用而在体细胞生长中起作用。GH也在胎盘中产生。胎盘GH在调节母体和胎儿代谢中起作用。垂体生长激素是从垂体前叶分泌的生长激素。生长激素(GH)的释放由从下丘脑分泌的生长激素释放激素(GHRH)调节(参见例如Ascoli et al.(1996)“Adenohypophyseal hormones and their hypothalamic releasing factorsGoodman&Gilman′s
The Pharmacological Basis of Therapeutics,Hardman et al.eds)pp.1363-1382,McGraw-Hill,New York)。生长激素以阵发式和脉冲式放射释放(参见例如Brook et al.Endocrinol.Metab.Clin.North Am.21:767-782(1992))。
生长激素已在人和其它哺乳动物中被研究。已经在人类中研究了对GH的药动学应答。这些应答分为两类:1)立即应答,包括产生胰岛素样生长因子I和II(IGF-1和IGF-2),刺激脂肪组织中的甘油三酯水解以及刺激肝糖输出;2)hGH的中间应答,包括合成代谢和IGF-I介导的生长促进,包括软骨发生、骨骼生长和软组织生长(Ascoli et al.,1996).
肝是产生IGF-I的主要器官。肝IGF-I诱导通过增加IGF-ImRNA转录而依赖于hGH(参见例如,Mathews LS,Norstedt G andPalmiter RD Proc.Natl.Acad.Sci.USA 83:9343-9347(1986))。循环中的IGF-I的主要形式是与40-kDa胰岛素样生长因子结合蛋白3(IGFBP-3)以及85-kDa酸不稳定亚基结合的150-kDa复合物(参见例如,Baxter et al.J.Biol.Chem.264:11843-11848(1989))。血浆中循环的这一复合物将IGF-I从肝转运至作用位点以促进生长。整体上,GH在调节IGF-I、IGFBP-3和不耐酸亚基中起主要作用(参见例如,Kupfer et al.J.Clin.Invest.91:391-396(1993))。
1.生长激素结构
GH多肽是由螺旋束组成的单结构域蛋白质,所述螺旋束由4个α螺旋(A、B、C和D)组成,这四个α螺旋由4个螺旋间环(AB、BC、CD和DE)连接在一起。二硫键可以进一步稳定GH结构。例如,人类生长激素同种型具有两个二硫键(成熟链的半胱氨酸53-165和182-189)。这两个二硫键连接环AB和DE。二硫键的存在看起来对稳定性和与受体的有效结合有贡献。
来自不同物种的生长激素多肽具有保守的氨基酸序列(参见例如,Abdel-Meguid et al.Proc.Natl.Acad.Sci.USA 84:6434-37(1987))。特别是在蛋白质的α螺旋区的序列保守性特别高。生长激素多肽典型地约为200个氨基酸。成熟人类生长激素含有191-222个氨基酸,在除去26个氨基酸的信号肽之后分子量为22kDa。通常,GH作为一个更长的多肽(即前体多肽)产生,其在裂解信号序列之后成熟为一较短的多肽;成熟GH约为191个氨基酸。
生长激素是异源蛋白质并且以长度为180、191、202、216、217和222个氨基酸的形式存在。异源生长激素多肽的例子包括但不限于具有SEQ ID NO:1和712-715所示的氨基酸序列的蛋白质。在一个实施方案中,生长激素多肽的长度为191、192、193、194、195、196、197、198、199、200、201、202、203、204、205、206、207、208、209、210、211、212、213、214、215、216、217、218、219、220、221或222个氨基酸。在人类中,编码垂体(hGH-N)和胎盘(hGH-V)生长激素的基因编码217个氨基酸的多肽。还存在GH的其它同种型,它们例如由可变剪接而生成。例如存在214个氨基酸的垂体GH的同种型。GH的胎盘和垂体同种型在整个蛋白范围内有序列相似性。例如,成熟GH胎盘同种型(GenBankAccession No.NP 002050.1)和成熟GH垂体同种型(GenBankAccession No.NP 000506.2)蛋白质序列的比较显示这两个成熟GH蛋白质序列之间有13个氨基酸的差异。
2.与生长激素受体的相互作用
生长激素与其受体之间的界面可以用分辨GH/受体复合物的晶体结构和诱变研究而鉴定。复合物是由一个配体分子(GH)和两个受体分子形成的。结果,在GH分子上有两个与受体的相互作用位点。复合物的表观解离常数在纳摩尔范围(Kd~0.3nM),但是这两个位点不以相同亲和性相互作用。一个相互作用位点(称为位点1)有高亲和性,另一个位点(称为位点2)有低亲和性。这一亲和性的不同反映在不同的界面表面积中(位点1为1300_2,位点2为850_2)。
只有几个位于界面的GH残基看起来对于与受体的结合是重要的。这些残基主要是疏水性残基。将这些疏水性残基突变为丙氨酸对于激素/受体复合物的结合常数(kon)没有作用,但是却增加其解离常数(koff)。因此,这些突变通过加速复合物解离而导致低亲和性。这些诱变结果表明激素与受体的结合是通过快速碰撞这一简单现象进行,而结合位点处的疏水性残基侧链的作用是当激素到达受体时保持激素结合。特定的极性残基、带电和不带电残基对于GH与其受体的结合也是重要的。这些极性残基保持结合位点的hGH的疏水性残基和受体的正确堆积(packing)。
3.作为生物药物的生长激素
生长激素作为治疗剂被给药。例如,在人类中,GH用作治疗剂以治疗儿童和成人生长缺陷、特纳氏综合征以及AIDS消瘦等疾病。用GH治疗是熟知的疗法。接受GH对象是通常以日为基础非常频繁重复应用该药物的对象。例如,GH替代通常每日皮下(s.c.)注射给予(参见例如Albertsson-Wikland et al.Acta.Paediatr.Scand.75:89-97(1986))。必须频繁重复应用GH是因为其在血流中和储存条件下不稳定导致的。
因此,在血清和体外(例如在生产、纯化和储存条件)的改善的GH稳定性(半衰期)将改善其作为药物的用途和有效性。因此,本发明提供了GH变体的突变变体,它们展示通过对蛋白酶的抗性和/或增加的热耐受性而评价的改善的稳定性,由此具有增加的蛋白质半衰期。展示改善的稳定性的突变变体具有在血流中和/或储存条件下的增加的稳定性。
C.修饰生长激素的举例性方法
本发明提供了修饰的生长激素蛋白质。修饰的生长激素(本文中也称为变体)与未修饰的生长激素相比稳定性增加。增加稳定性(例如蛋白质的体外半衰期)可以降低保持血清中的足够药物浓度所需的注射频率,由此使得i)被治疗对象特别是人类对象的舒适度更高和更可接受,ii)达到相当的生物学作用所需的剂量降低,以及iii)其结果是(剂量依赖性)副作用减弱。
GH的增加的稳定可以例如通过破坏蛋白酶靶残基或序列和/或(ii)增加蛋白质的热耐受性而实现。增加稳定性的GH修饰可以在保持生物学活性相对于未修饰的或野生型GH未变化的同时而实现。本领域已知的任何方法均可以用于产生修饰的生长激素蛋白。在本文描述的方法中,使用2D扫描诱变的方法选择修饰(参见例如,WO2004/022747和WO 2004/022593)。
蛋白酶或肽酶催化共价肽键的水解。丝氨酸蛋白酶参与机体的各种功能,包括凝血、炎症和原核和真核生物中的消化酶类。丝氨酸蛋白酶是序列特异性的。蛋白酶活化级联控制凝血和补体,而其它蛋白酶参与信号传导途径、酶活化和补体细胞或细胞外区室中的降解功能。
丝氨酸蛋白酶包括但不限于胰凝乳蛋白酶、胰蛋白酶、弹性蛋白酶、基质金属蛋白酶如明胶酶B和明胶酶A、NS3、弹性蛋白酶、因子Xa、粒酶B、凝血酶、胰蛋白酶、纤溶酶、尿激酶、tPA和PSA。胰凝乳蛋白酶、胰蛋白酶和弹性蛋白酶由胰腺腺泡细胞合成,分泌在小肠中并且负责催化肽键水解。这三种酶在结构上类似,如它们的X光结构所示。这些消化性丝氨酸蛋白酶的每一个靶向多肽链的不同区域,其基于裂解位点周围的氨基酸残基和侧链。丝氨酸蛋白酶的活性位点呈裂缝形式,该裂缝是多肽底物结合的地方。氨基酸残基从多肽底物的N至C端(Pi,...,P3,P2,P1,P1′,P2′,P3′,...,Pj)以及它们各自的结合亚位点(Si,...,S3,S2,S1,S1′,S2′,S3′,...,Sj)标记。裂解在P1和P1’之间被催化。胰凝乳蛋白酶负责裂解两侧为大体积疏水氨基酸残基的肽键。特别的残基包括苯丙氨酸、色氨酸和酪氨酸,其适合合适的疏水口袋。胰蛋白酶负责裂解两侧为带正电氨基酸残基的肽键。代替胰凝乳蛋白酶的疏水口袋,在口袋的后面(back)存在天冬氨酸残基。其然后可以与带正电的残基如精氨酸和赖氨酸相互作用。弹性蛋白酶负责裂解两侧为小中性氨基酸残基如丙氨酸、甘氨酸和缬氨酸的肽键。胰蛋白酶和胰凝乳蛋白酶中的口袋此时与缬氨酸和苏氨酸配合,使得其单纯收缩,可以容纳这些较小的氨基酸残基。
丝氨酸蛋白酶在原核和真核生物中遍在,并且起多种重要的生物学功能如造血、纤维蛋白溶解作用、补体形成和膳食蛋白消化。丝氨酸蛋白酶包括基质金属蛋白酶如明胶酶B(基质金属蛋白酶9(MM9))和明胶酶A。对其抗性增加的蛋白酶包括例如在体液和组织中存在的蛋白酶,如包括但不限于唾液、血液、血清、小肠、胃、血液、细胞裂解物、细胞和其它中的蛋白酶。它们包括全部类型的蛋白酶。
原则上,对于基于诱变的蛋白质定向进化描述了几种通用方法。这些方法单独或组合可以用于修饰多肽如GH以实现增加的构象稳定性和/或对蛋白酶解的抗性。这些方法包括随机诱变,其中起始蛋白质序列中的氨基酸被全部(或一组)20个氨基酸在补体氨基酸位置进行单置换或多个置换,在相同分子上同时进行。另一种方法限制随机诱变导入全部20个氨基酸或DNA偏倚残基(DNA-biasedresidue)。偏向是基于DNA序列而非蛋白质序列,分别以随机或半随机方式,在蛋白质的事先已知参与被“进化”的生物学活性中的限制或预定区域。另外,包括1D-扫描、2D-扫描和3D-扫描的合理诱变可以单独使用或组合使用以构建修饰的hGH变体。
1.1D扫描(“合理诱变”)
合理诱变也称为1D-扫描,是一个两步方法并且在共同在审美国专利申请系列号10/022,249(U.S.申请公开No 2003-0134351-A1)中描述。简而言之,在第一步中,进行全长氨基酸扫描,其中起始蛋白质如hGH(SEQ ID NO:1)中的每一个氨基酸逐一被指定的参考氨基酸(例如丙氨酸)置换。一次在每一蛋白质分子上仅置换一个氨基酸。这些氨基酸位置被称为HIT。在第二步,产生分子的新集合,从而每个分子之间的区别是在步骤1中鉴别的各个HIT位置存在的氨基酸。在步骤1中鉴别的每一HIT位置引入全部20个氨基酸(其余19个);而每个分子原则上含有一个并且仅有一个氨基酸置换。这些新产生的导致蛋白质活性的希望的改变(如改进)的突变体被称为LEAD。这些方法使得能在蛋白质的未预测区域的新的未预测氨基酸序列,以产生展示与起始蛋白质相比所希望的改变的活性的蛋白质。另外,因为对用于第二步的靶区域(HIT和周围氨基酸)的选择基于在第一步获得的有关活性的实验数据,因此无需关于蛋白质结构和/或功能的在先知识。
2.3D扫描
3D-扫描在共同在审美国公开申请No.2004-0132977-A1和U.S.申请系列号10/685,355和公开的PCT申请WO 2004/022747和WO2004/022593中描述,其是另外一种基于鉴别用于诱变的潜在靶位点(is-HIT)的蛋白质合理进化方法。该方法利用结构相关的蛋白质之间的蛋白质主链折叠模式的比较,而不考虑被比较的蛋白质的序列。一旦在感兴趣蛋白质上鉴别了结构相关的氨基酸位置,则可以采用合适的氨基酸置换标准如PAM分析来鉴别用于构建和扫描的候选LEAD。
3.2D-扫描(限制合理诱变,restricted rational mutagenesis)
用于蛋白质合理进化的2D-扫描(或称限制合理诱变)方法(参见共同在审美国专利申请系列号10/685,355和美国公开申请No.US-2004-0132977-A1和公开的国际申请WO2004022593和WO2004022747)是基于在两维上的扫描。第一维是沿蛋白质序列的氨基酸位置,用以鉴别is-HIT靶位置。第二维是扫描所选择的用以置换特定is-HIT氨基酸位置的氨基酸类型。本文提供的2D-扫描方法的优点是至少一个、典型地是氨基酸位置和/或置换氨基酸可以被限制,从而蛋白质主链上少于全部氨基酸被选择用于氨基酸置换;和/或少于全部其余19个可利用的用于置换原始的如天然的氨基酸的氨基酸被选择用于置换。
基于i)有待进化的特定蛋白质性质(即对蛋白酶解的抗性),ii)蛋白质的氨基酸序列,和iii)各个氨基酸的已知性质,in silico选择蛋白质序列上的一些靶位置作为″is-HIT靶位置″。is-HIT靶位置数在合理范围内尽可能地大,以便包括用于待进化的特定特征的所有合理可能的靶位置。特别地,在有限数目的is-HIT靶位置被选择用于置换的实施方案中,被选择来置换被优化的特定蛋白质的is-HIT靶位置的氨基酸可以是其余19个氨基酸全部,或者更经常地是一更有限的组,其包含预期对蛋白质活性有希望的作用的选择氨基酸。在另一个实施方案中,如果使用有限数目的置换氨基酸,则蛋白质主链上的所有氨基酸位置可以被选择作为is-HIT靶位置用于氨基酸置换。然后通过置换蛋白质主链上的特异is-HIT靶位置的单氨基酸残基而进行诱变(例如“一个接一个”,如在可寻址阵列中),以便所产生的每个突变体是每个单诱变反应的单一产物。设计突变DNA分子,通过诱变产生并逐个克隆在如可寻址阵列中,以便它们互相之间物理分离,并且每一个是一个独立诱变反应的单一产物。衍生自突变DNA分子集合的突变蛋白质分子也是互相之间物理分离的,如通过格式化在可寻址阵列中。因此,产生了多个突变蛋白质分子。每个突变蛋白质含有在仅一个is-HIT靶位置的单氨基酸置换。然后在蛋白质表达和测量合适活性后,在每个蛋白质突变分子上逐个进行活性评价。实施这一方法的一个例子见实施例所述,其中产生突变hGH分子。
这些新产生的导致改变的、典型地改善的靶蛋白活性的蛋白质被称为LEAD。这一方法依赖于对特定活性(如增加的对蛋白酶解的抗性)的蛋白质改进的间接检索,其基于一次在一个氨基酸位置或在另一个实施方案中在有限数目的氨基酸位置的氨基酸置换和序列变化。其结果是,鉴别和分离了表现优于(在特定靶活性或其它活性方面)或不同于起始蛋白质的优化的蛋白质,它们在蛋白质的一些区域具有修饰的氨基酸序列。
使用对GH的2D-扫描产生了包括改善的对蛋白酶解的抗性和改善的热耐受性在内的在蛋白质稳定性方面改善的变体。为实现这类修饰,使用hGH的in silico分析选择氨基酸位置。
a.鉴别in-silico HIT
用于蛋白质的定向进化的2D-扫描方法包括沿蛋白质序列鉴别和选择(使用in silico分析)预期直接或间接参与被进化特征的特异的氨基酸和氨基酸位置(本文中称为is-HIT)。如所指明的,提供的2D-扫描方法包括下列两个步骤。第一步是蛋白质的靶氨基酸序列的in silico检索以鉴别潜在地可以是被进化的活性的靶的所有可能的氨基酸位置。这例如通过使用任何已知的标准软件评价蛋白质上氨基酸残基对待被改变的性质的作用而实现。in silico分析的细节取决于待被修饰的性质。
鉴别后,这些氨基酸位置或靶序列被称为″is-HIT″(in silicoHIT)。In silico HIT被定义为那些潜在地参与“进化的”特征如增加对蛋白酶解的抗性或热耐受性的氨基酸位置(或靶位置)。在蛋白质序列中的所有氨基酸位置中区分is-HIT除基于蛋白质二级或三级结构之外还可以基于每一位置的氨基酸类型。In silico HIT构成突变体分子集合,从而代表潜在地参与进化的特征中的所有可能的氨基酸、氨基酸位置或靶序列。在这一阶段不进行氨基酸或氨基酸位置中的强理论区分。In silico HIT位置分布在蛋白质序列全长中。在靶hGH蛋白上一次只置换单个或有限数目的is-HIT氨基酸。
可以分析各种参数以确定蛋白质上的一特定氨基酸是否可能参与进化的特征中,典型地使用有限数目的起始前提(典型地不超过2个)确定in silico HIT。例如,如本文所述,为增加hGH的热耐受性,第一个条件是与分子的热耐受性相关的氨基酸性质,如其在能参与分子稳定化的化学键中的潜在参与。第二个前提典型地与那些氨基酸在蛋白质结构上的特异位置。
在根据本文提供的方法鉴别is-HIT的第一步中,逐个考虑蛋白质序列的每个氨基酸以评价其是否是is-HIT的候选者。这一检索一个接一个地进行,而确定氨基酸是否被认为是is-HIT的候选者的决定是基于(1)氨基酸类型;(2)在蛋白质和蛋白质结构中的位置,如果知道的话,以及(3)氨基酸与其在序列和空间中邻近的氨基酸之间预测的相互作用。
针对与蛋白质稳定性相关的若干hGH性质,鉴别了is-HIT。这些性质包括1)增加螺旋之间的盐水(极性)相互作用;2)增加螺旋之间的H键相互作用,以及3)除去蛋白酶敏感位点。
b.鉴别置换氨基酸
一旦选择了is-HIT靶位置,下一步是鉴别置换每一is-HIT位置的原始如天然氨基酸以改变被进化的特定特征的活性水平的那些氨基酸。用于置换每一is-HIT位置的原始如天然氨基酸的一套置换氨基酸可以是不同的并且针对特定的is-HIT位置是特异的。置换氨基酸的选择要考虑保留生理化学性质如关键(例如催化的结合等)残基的疏水性、电荷和极性并改变蛋白质的其它一些性质(即蛋白质稳定性)的需要。可用于置换特定is-HIT靶位置的其余19个非天然(或非原始)氨基酸的置换氨基酸数从1直至约19并且是其间的任意数,这取决于特定修饰的性质.
选择置换氨基酸(本文中也称为″replacement amino acids″)的各种方法是本领域熟知的。蛋白质化学家已确定在来自不同物种的相关蛋白质中通常发生某些氨基酸取代。由于具有这些取代的蛋白质仍然有功能,因此这些被取代的氨基酸与蛋白质结构和功能是相容的。通常,这些取代是取代化学相似的氨基酸,但是也可以发生其它类型的变化,尽管这相对较稀有。
了解在大量蛋白质中最常见和最不常见的变化类型可以有助于预测针对任何蛋白质序列集合的对比和氨基酸取代。氨基酸取代矩阵用于这一目的。有许多矩阵可利用。关于这类矩阵的详细描述可见于共同在审美国专利申请系列号10/685,355和美国申请公开号US-2004-0132977-A1和公开的国际申请WO 2004/022593 and WO2004/022747,这些文献全文引入本文(当允许时)。这类矩阵也是本领域已知的和可或的,例如在下面所列的参考文献中。
在氨基酸取代矩阵中,氨基酸被水平和垂直列出,每一矩阵位置被填上一分值,该分值反映出在相关蛋白质序列对比中一个氨基酸与另一个之间配对的经常性如何。氨基酸“A”变化为氨基酸“B”的概率被假定为与“B”变化为“A”的逆概率是相同的。做出这一假定是因为对于任何两个序列,在系统进化树中的祖先氨基酸通常是未知的。另外,置换可能性取决于两个氨基酸发生频率的乘积以及取决于它们的化学和物理相似性。这一模型的预测是氨基酸频率不随进化时间而变化(Dayhoff et al.,Atlas of Protein Sequenceand Structure,5(3):345-352,(1978))。几种举例性氨基酸取代矩阵包括但不限于block substitution matrix(BLOSUM)(Henikoff et al.,Proc.Natl.Acad.Sci.USA,89:10915-10919(1992)),Jones(Jones et al.,Comput.Appl.Biosci.,8:275-282(1992)),Gonnet(Gonnet et al.,Science,256:1433-1445(1992)),Fitch(J.Mol.Evol.,16(1):9-16(1966)),Feng(Feng et al.,J.Mol.Evol.,21:112-125,(1985)),McLachlan(J.Mol.Biol.,61:409-424(1971)),Grantham(Science,185:862-864(1974)),Miyata(J.Mol.Evol.,12:219-236(1979)),Rao(J.Pept.Protein Res.,29:276-281(1987)),Risler(J.Mol.Biol.,204:1019-1029(1988)),Johnson(Johnsonet al.,(J.Mol.Biol.,233:716-738(1993))和Point Accepted Mutation(PAM)(Dayhoff et al.Atlas Protein Seq.Struct.5:345-352(1978))。
in silico进行的上述两个步骤的结果是:(1)鉴别了是诱变靶的氨基酸位置(称为is-HIT);以及(2)鉴别了用于置换is-HIT的原始如天然氨基酸的置换氨基酸,提供了预期与天然分子行为不同的候选LEAD突变分子集合。它们被测定是否具有所希望的优化的(或改善的或改变的)生物学活性。
c.构建修饰的多肽和生物学测定
一旦如上所述选择了is-HIT,则导入置换氨基酸。突变蛋白质典型地用重组DNA方法制备并在合适的生物学测定中评价被优化的特定生物学活性(特征)。制备突变蛋白质的举例性方法是用本领域熟知的方法诱变原始的如天然的基因。如在可寻址阵列中一个接一个产生突变分子,以便所产生的每个突变体是每个单一独立诱变反应的单一产物。如在可寻址阵列中单独进行各个诱变反应,由此它们互相物理分离。一旦制备编码各个突变蛋白质的核酸分子集合群,一个接一个将每个单独导入合适的细胞中以产生相应的突变蛋白质。这也可以在例如可寻址阵列中进行,在其中编码各自突变蛋白质的每个核酸分子集合被导入限制在一个分离位置中如在多孔微滴板的孔中的细胞中。每个突变蛋白质逐个进行表型鉴定并用适用于被优化的特征(即被进化的特征)的测定定量评价行为表现。同样这一步骤可以在可寻址阵列中进行。与原始如天然的分子相比展示所希望的增加的或降低的行为表现的突变体被鉴别并被称为LEAD。从该方法开始产生突变体DNA分子直至读出和分析行为表现结果,如从在可寻址阵列中的自身地址逐个产生、生产和分析每个候选LEAD突变体。这一方法适于自动化。
D.修饰的生长激素多肽
本发明提供的是通过增加对蛋白酶解的抗性和/或通过增加热耐受性而增加多肽的稳定性和半衰期的方法。本发明提供了修饰多肽如生长激素以增加对蛋白酶(血液、血清、胃肠道等的蛋白酶)导致的蛋白酶解的抗性的方法,其中修饰的多肽展示增加的体外和/或体内半衰期。本发明提供了将蛋白酶解酶与肽抑制剂接触由此抑制蛋白酶(血液、血清、胃肠道等的蛋白酶)的生物学活性的方法。接触的结果是,对蛋白酶解敏感的多肽展示增加的体外或体内半衰期。本发明提供了修饰多肽以增加对蛋白酶所致的蛋白酶解的抗性的方法,以及将蛋白酶解酶与肽抑制剂接触由此抑制蛋白酶的生物学活性的方法。本发明还提供了由所述方法的产生的修饰的多肽。本发明提供了展示由对蛋白酶的抗性和/或增加的热耐受性评价的改善的稳定性的修饰的生长激素多肽;展示这些性质的修饰的多肽由此具有增加的蛋白质体外或体内半衰期。
本发明提供了展示由对蛋白酶的抗性和/或增加的热耐受性评价的改善的稳定性的修饰的GH多肽;展示这些性质的修饰的多肽由此具有增加的蛋白质体外或体内半衰期。
本发明提供了展示由对蛋白酶的抗性和/或增加的热耐受性评价的改善的稳定性的修饰的(变体、突变体等)GH(在本文中也称为修饰的GH多肽)。这种修饰的GH多肽可以具有增加的蛋白质体外半衰期(例如在生产、纯化和储存过程中)和体内半衰期。在一个实施方案中,本发明提供的修饰的GH多肽赋予至少一种相当的(comparable)生物学活性。例如,修饰的GH多肽与未修饰的和/或野生型天然GH多肽相比赋予至少一种由GH特异性增殖活性评价的相当的生物学活性。
本发明提供的修饰的GH多肽包括人GH变体。本发明提供的修饰的GH多肽与SEQ ID No;1的氨基酸序列相比已被修饰。hGH多肽可以是任何人类组织或细胞类型来源的。人类GH多肽包括例如SEQ ID NO:1,712和713所示的多肽序列。本发明提供的修饰的GH多肽还包括非人来源的GH变体。例如,修饰的GH多肽可以是哺乳动物GH如牛、大鼠、兔、绵羊、灵长类、马、猪、猴、狒狒、长臂猿、猕猴、大猩猩、猩猩和黑猩猩GH的变体。修饰的GH多肽也可以包括是不同GH序列的杂种的多肽,以及从现有技术已知的GH序列构建的合成GH序列。
本发明提供的修饰的GH多肽改变了赋予GH稳定性的细胞因子的特异结构特征。本发明提供的修饰的GH多肽包括具有增加的对蛋白酶解的抗性和/或增加的热耐受性的变体。因此,本发明提供的修饰的GH多肽提供了具有如下优点的GH,即维持血清中足够的药物浓度所需的注射频率降低,由此导致例如对象的舒适度更高和更可接受,达到相当的生物学作用所需的剂量降低,以及副作用减弱。
通过氨基酸置换可以在GH中进行结构修饰以增加GH的构象稳定性。这类修饰包括增加GH的构象稳定性同时改善或维持所需的生物学活性(例如细胞增殖活性)的那些修饰。这些修饰可以产生具有由对蛋白酶的抗性和/或增加的热耐受性评价的改善的稳定性的GH变体。这些修饰的GH多肽展示与天然GH相比的增加的蛋白质体外和/或体内半衰期。这些修饰包括破坏对蛋白酶解敏感的靶氨基酸序列,修饰疏水patches以增加与溶剂的极性相互作用,以及增加GH特定螺旋之间的极性相互作用。
GH的结构修饰包括组合在GH序列内的不同位置的一个、两个或多个氨基酸置换以增加整个GH的稳定性。这种组合可以用于改善由对蛋白酶的抗性和/或增加的热耐受性评价的稳定性。例如,一或多个类别中的两种或多种修饰可以被组合,其中这些类别选自例如对蛋白酶解敏感的靶序列的破坏,修饰疏水patches以增加与溶剂的极性相互作用,增加螺旋之间的极性相互作用以及增加螺旋之间的相互作用。另外,这些类别中的一种或多种修饰可以与一或多种任何已知类型的修饰组合以增加GH稳定性,例如,去除GH中的蛋白酶敏感位点的修饰与增加热耐受性的任何已知类型的修饰组合。
在变体中,本发明还提供了与天然或野生型GH相比具有两个或多个修饰的修饰的GH多肽。在一个例子中,hGH是修饰的垂体hGH。修饰的GH多肽包括具有2、3、4、5、6、7、8、9、10、11、12或更多个修饰的位置的那些多肽。所述两个或多个修饰可以包括相同性质的两个或多个修饰,例如修饰hGH热耐受性的两个修饰或者修饰对蛋白酶抗性的两个修饰。在另一个实施方案中,所述两个或多个修饰包括各自对GH稳定性有贡献的性质的组合。例如,修饰的GH多肽可以包括一或多个改变GH热耐受性的修饰以及一或多个去除蛋白酶敏感位点的修饰。在一个实施方案中,是在成熟生长激素的氨基酸序列中有一或多个氨基酸置换的修饰的生长激素多肽,其中如果位置9被置换,则置换氨基酸不是脯氨酸;如果位置13被置换,则置换氨基酸不是缬氨酸;如果位置14被置换,则置换氨基酸不是丝氨酸;如果位置54被置换,则置换氨基酸不是脯氨酸;如果位置56被置换,则置换氨基酸不是天冬氨酸;如果位置64被置换,则置换氨基酸不是甲硫氨酸;如果位置65被置换,则置换氨基酸不是缬氨酸或丙氨酸;如果位置66被置换,则置换氨基酸不是谷氨酰胺或赖氨酸;如果位置92被置换,则置换氨基酸不是亮氨酸;如果位置120被置换,则置换氨基酸不是精氨酸;如果位置126被置换,则置换氨基酸不是精氨酸;如果位置129被置换,则置换氨基酸不是苏氨酸;如果位置133被置换,则置换氨基酸不是组氨酸或精氨酸;如果位置134被置换,则置换氨基酸不是亮氨酸;以及如果位置140被置换,则置换氨基酸不是天冬酰胺。在超过一个is-HIT位点携带置换并且展示改善的稳定性的GH变体被称为super-LEAD。GH super-LEAD与野生型或未修饰的GH相比可以例如含有2、3、4、5、6、7、8、9、10或多于10个氨基酸变化。
本发明提供了上述任一种修饰的生长激素,其中与未修饰的生长激素相比被置换的位置数是2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19或20个。在一个实施方案中,本发明提供的修饰的生长激素包括在一或多个如下位置的氨基酸置换:具有SEQ ID NO:1的氨基酸序列的成熟人类生长激素的位置56、59、64、65、66、88、92、94、101、129、130、133、134、140、143、145、146、147、183和186。在一个实施方案中,所述位置包括E56、P59、R64、E65、E66、E88、F92、R94、L101、E129、D130、P133、R134、K140、Y143、K145、F146、D147、R183和E186。在一个实施方案中,位置被如下置换:用Q,N和H任一个置换E、用S或A置换P、用H或Q置换R、用I或V置换L或F、用Q或N置换K或D、以及用H或I置换Y。例如,这类置换包括E56Q、E56N、E56H、P59S、P59A、R64H、R64Q、E65Q、E65N、E65H、E66Q、E66N、E66H、E88Q、E88N、E88H、F92I、F92V、R94H、R94Q、L101V、L101I、E129Q、E129N、E129H、D130Q、D130N、P133S、P133A、R134H、R134Q、K140Q、K140N、Y143H、Y143I、K145Q、K145N、F146I、F146V、D147Q、D147N、R183H、R183Q、E186Q、E186N和E186H。
1.通过去除蛋白酶解位点而实现的对蛋白酶解的增加的抗性
对于治疗蛋白质如GH的感兴趣修饰是增加构象稳定性的修饰。增加构象稳定性可以通过增加蛋白质对蛋白酶解的抗性而实现。这种稳定性增加可以包括增加蛋白酶抗性(及因此的蛋白质半衰期)而同时保持所需的生物学活性。这类变化对于生产保持时间更长的治疗蛋白质是有用的。
a.通过去除蛋白酶解位点而修饰的生长激素多肽的性质
本发明提供了具有增加的体外或体内稳定性的GH多肽的修饰,其通过增加修饰的GH对蛋白酶解的抗性而导致。在修饰的GH多肽中,本发明提供了如下修饰的GH变体:1)增加螺旋之间的盐水(极性)相互作用;2)增加螺旋之间的H键相互作用以及3)去除蛋白酶敏感位点。
在一个实施例中,稳定性增加的修饰的GH多肽是人类GH多肽。使用2D-扫描方法学鉴别了hGH上的导致当用蛋白酶(血液、血清、肠道蛋白酶)、血液裂解物或血清攻击时稳定性增加的氨基酸变化。本文中预期增加对蛋白酶、血液裂解物或血清的蛋白质稳定性能使特定蛋白质分子体内半衰期延长,因此降低给予对象的必要注射的频率。在一个实施例中,修饰的hGH的生物学活性在测定修饰的GH当被加至合适细胞时刺激细胞增殖的能力的测定中评价。在测量生物学活性之前,在不同保温或注射猴时间hGH分子可以暴露于血液、血清或肠道蛋白酶(体外测定)或血清/肠道(在小鼠中的体内测定)。测得的生物学活性相应于暴露于蛋白酶解混合物后的残留生物学活性。修饰的GH的生物学活性可以与未修饰的GH进行比较以作为所述修饰对蛋白酶稳定性和生物学活性的作用的测量。在一个实施例中,未修饰的GH是野生型天然GH。在另一个实施例中,未修饰的GH是用作起始材料以引入进一步修饰的GH变体形式。修饰的GH也可以与任何已知的GH多肽在这类测定中进行比较以比较蛋白酶敏感性和/或生物学活性。另外,现有技术中已知的任何评价蛋白酶稳定性、蛋白酶抗性和敏感性以及GH生物学活性的测定可以被用于评价本发明的修饰的GH多肽。
本发明提供了保持所需生物学活性基本未改变并且已经对血液、血清或肠道蛋白酶的消化的敏感性下降并因此展示更长的循环半衰期的GH分子。这类GH分子包括具有对于治疗性应用足够的生物学活性的修饰的GH多肽。
本发明提供了具有增加的蛋白酶抗性的修饰的GH多肽。在举例性的实施方案中,本发明提供的GH突变体的体外或体内半衰期(血清稳定性)当与在人血、人血清或含有一或多种蛋白酶的体外混合物中的天然GH的半衰期相比时增加了至少10%、至少20%、至少30%、至少40%、至少50%、至少60%、至少70%、至少80%、至少90%、至少100%、至少150%、至少200%、至少250%、至少300%、至少350%、至少400%、至少450%、至少500%或更多。在其它实施方案中,本发明提供的GH突变体的体外或体内半衰期(血清稳定性)当与在人血、人血清或含有一或多种蛋白酶的体外混合物中的天然GH的半衰期相比时增加了至少6倍、7倍、8倍、9倍、10倍、20倍、30倍、40倍、50倍、60倍、70倍、80倍、90倍、100倍、200倍、300倍、400倍、500倍、600倍、700倍、800倍、900倍、1000倍或更多倍。在一个举例性实施方案中,这类GH变体是通过修饰人类GH多肽而产生。在一个这种举例性实施方案中,GH变体是通过修饰人垂体GH多肽而产生。
b.通过去除蛋白酶解位点而修饰的生长激素多肽的产生
在一个通过去除蛋白酶解位点而产生稳定性增加的变体的实施例中,修饰了垂体hGH。设计抗蛋白酶解的hGH突变体的第一步包括沿蛋白质序列鉴别对蛋白酶解敏感的位点。基于所考虑的选定蛋白酶列表(表2)和本文描述的其它血液、血清和肠道蛋白酶,首先in silico确定hGH中可以被那些蛋白酶靶向的所有氨基酸和氨基酸序列的完整列表。蛋白酶靶(hGH序列上的氨基酸或氨基酸序列)被称为in silico HIT(is-HIT)。由于体内蛋白酶混合物的组成非常复杂,因此可以预期一给定蛋白质序列中的大多数残基均可以被靶向而被蛋白酶解。
设计抗蛋白酶解的hGH突变体的第二步包括鉴别合适的置换氨基酸,以便如果它们在is-HIT置换hGH中的天然氨基酸,则蛋白质将(i)变得对蛋白酶解有抗性以及(ii)引起与野生型hGH蛋白相当的生物学活性水平。置换氨基酸的选择必须考虑某些蛋白酶的广谱靶特异性以及需要保留生理化学性质如疏水性、GH中的关键(例如催化、结合等)残基的电荷和极性。
所谓的“Point Accepted Mutation”(PAM;Dayhoff et al.,1978(Figure 2))是2D-扫描方法的一部分。最初产生的用于对比蛋白质序列的PAM值以概率矩阵形式提供,其反映了氨基酸之间的进化距离。参考序列中的残基的“保守取代”是与相应的参考残基物理上和功能上相似的取代,即具有相似的大小、形状、电荷、化学性质包括形成共价或氢键以及其它相互作用的能力。保守取代显示出与“可接受的点突变”形式的PAM矩阵标准的最高分值的适合。PAM250矩阵用于2D-扫描框架以鉴别用于is-HIT的候选置换氨基酸,产生不影响蛋白质功能的保守突变。选择在PAM250矩阵中具有最高值、相应于“保守取代”或“可接受的点突变”的至少2个氨基酸用于在每个is-HIT进行置换。要明确避免用半胱氨酸残基置换氨基酸,因为这种变化会潜在地导致形成分子间二硫键。
简而言之,使用算法PROTEOL(从
infobiogen.fr和bioinfo.hku.hk/services/analyseq/cgi-bin/proteol_in.pl在线获得),沿191个氨基酸的hGH蛋白(SEQ ID NO:1)建立了可以被识别为表2中的蛋白酶的底物的残基列表(图1)。该算法基于蛋白酶列表、它们的蛋白酶解特异性以及输入的多肽氨基酸序列产生了蛋白酶解消化图谱。
表2
缩写 | 氨基酸位置 | 蛋白酶或化学处理 |
AspN | D | 内切蛋白酶Asp-N |
Chymo | (F,W,Y,M,L)~P | 胰凝乳蛋白酶 |
Clos | R | 梭菌蛋白酶 |
CnBr | M | 溴化氰 |
IBzO | W | IodosoBenzoate |
Myxo | K | Myxobacter |
NH2OH | NG | 羟基胺 |
pH2.5 | DP | pH 2.5 |
ProEn | P | 脯氨酸内肽酶 |
Staph | E | 葡萄球菌蛋白酶 |
Tryp | (K,R)~P | 胰蛋白酶 |
TrypK | K~P | 胰蛋白酶(Arg blocked) |
TrypR | R~P | 胰蛋白酶(Lys blocked) |
表2显示了作为使用各种所选择的蛋白酶和化学处理的蛋白酶解的靶的一些氨基酸位置的in silico鉴别。
鉴别is-HIT并产生LEAD以获得对蛋白酶解的更高的抗性的hGH。在每一is-HIT位置的天然氨基酸和用于获得增加的对蛋白酶解的抗性的置换氨基酸可以包括但不限于用H和/或Q置换R;用H,Q和/或N置换E;用Q和/或N置换K;用N和/或Q置换D;用I和/或V置换M;用A和/或S置换P;用I和/或H置换Y;用I和/或V置换F;用H和/或S置换W;用I和/或V置换L(见表3)。is-HIT和LEAD可以包括在对蛋白酶解敏感的特定区域的修饰。
在一个实施方案中,选择用于修饰的区域包括在相应于选自SEQ ID NO:1的氨基酸1-12、14-26、29 to 53、57、58、60-63、67-78、80-84、86、87、89、91、93、95-100、102-113、115-128、131、132、135-139、141、142、144、148-160、162-182、185以及187-191的成熟人类生长激素的位置的区域中的一或多个氨基酸修饰。在另一个实施方案中,被修饰的位置相应于一或多个、例如2、3、4、5、6、7、8、9、10、11、12,13、14、15、16、17、18、19、20个或更多个位置。在一个实施方案中,GH被修饰以包括与未修饰的生长激素相比的一或多个单氨基酸置换,其中置换位置不是相应于SEQ ID NO:1的位置13、27、28、54-56、59、64-66、79、85、88、90、92、94、101、114、129、130、133、134、140、143、145-147、161、183、184和186的位置。
例如,垂体hGH被修饰以在相应于选自SEQ ID NO:1的成熟人类生长激素的氨基酸位置1-12、14-26、29-53、57、58、60-63、67-78、80-84、86、87、89、91、93、95-100、102-113、115-128、131、132、135-139、141、142、144、148-160、162-182、185和187-191的氨基酸位置中的一或多个氨基酸修饰。在另一个实施方案中,胎盘hGH在这些相应位置被修饰。胎盘hGH与垂体hGH进行序列对比,选择相应于垂体hGH例如SEQ ID NO:1的位置的一或多个氨基酸位置。在另一个实施方案中,修饰非人来源的GH,例如牛、绵羊或猴GH。这种序列对比和位置选择可以用任何GH多肽进行,即将其与垂体hGH对比并选择相应的位置进行修饰。
在一个实施方案中,选择相应于垂体hGH的位置(is-HIT)并制备具有增加的对蛋白酶解的抗性的氨基酸置换(LEAD)。位置包括但不限于F1I/P2A、F1I/P2S、F1V/P2A、F1V/P2S、P5A、P5S、L6I、L6V、R8H、R8Q、L9I、L9V、F10I、F10V、D11N、D11Q、M14I、M14V、L15I、L15V、R16H、R16Q、R19H、R19Q、L20I、L20V、L23I、L23V、F25I、F25V、D26N、D26Q、Y28H、Y28I、E30Q、E30H、E30N、F31I、F31V、E32Q、E32H、E32N、E33Q、E33H、E33N、Y35H、Y35I、P37A、P37S、K38N、K38Q、E39Q、E39H、E39N、K41N、K41Q、Y42H、Y42I、F44I、F44V、L45I、L45V、P48A、P48S、L52I、L52V、F54I、F54V、E56Q、E56H、E56N、P59A、P59S、P61A、P61S、R64H、R64Q、E65Q、E65H、E65N、E66Q、E66H、E66N、K70N、K70Q、L73I、L73V、E74Q、E74H、E74N、L75I、L75V、L76I、L76V、R77H、R77Q、L80I、L80V、L81I、L81V、L82I、L82V、W86H、W86S、L87I、L87V、E88Q、E88H、E88N、P89A、P89S、F92I、F92V、L93I、L93V、R94H、R94Q、F97I、F97V、L101I、L101V、Y103H、Y103I、D107N、D107Q、Y111H、Y111I、D112N、D112Q、L113I、L113V、L114I、L114V、K115N、K115Q、D116N、D116Q、L117I、L117V、E118Q、E118H、E118N、E119Q、E119H、E119N、L124I、L124V、M125I、M125V、R127H、R127Q、L128I、L128V、E129Q、E129H、E129N、D130N、D130Q、P133A、P133S、R134H、R134Q、F139I、F139V、K140N、K140Q、Y143H、Y143I、K145N、K145Q、F146I、F146V、D147N、D147Q、D153N、D153Q、D154N、D154Q、L156I、L156V、L157I、L157V、K158N、K158Q、Y160H、Y160I、L162I、L162V、L163I、L163V、Y164H、Y164I、F166I、F166V、R167H、R167Q、K168N、K168Q、D169N、D169Q、M170I、M170V、D171N、D171Q、K172N、K172Q、E174Q、E174H、E174N、F176I、F176V、L177I、L177V、R178H、R178Q、R183H、R183Q、E186Q、E186H、E186N、F191I和F191V(见表4和SEQID NO:2-223)。关于这些突变体,第一个氨基酸(单字母缩写)相应于被置换的氨基酸,数字相应于在SEQ ID NO:1的hGH序列中的位置,第二个氨基酸(单字母缩写)相应于选择用于置换该位置的氨基酸的氨基酸。用于修饰的GH可以是任何GH,包括其它哺乳动物GH和胎盘GH。鉴别和修饰通过合适的序列对比估算的相应位置。
c.另外的修饰的生长激素多肽
在特定的实施方案中,也可以产生含有在一或多个is-HIT位点已被候选LEAD氨基酸置换的一或多个氨基酸的变体GH分子。在一或多个is-HIT携带一或多个突变并且展示改善的蛋白酶抗性的那些突变体蛋白质被称为LEAD(在一个is-HIT有一个突变)和supe-LEAD(在一个以上is-HIT有突变)。
在另外的实施方案中,展示改善的蛋白酶抗性的突变体分子LEAD和super-LEAD可以被进一步用赋予蛋白质稳定性的另外的修饰如本文描述的那些修饰而修饰。例如,蛋白酶抗性LEAD和super-LEAD可被修饰以含有如下突变:1)增加螺旋之间的盐水(极性)相互作用;2)增加螺旋之间的H键相互作用以及3)导致对蛋白酶解的抗性增加的其它修饰。
d.具有对蛋白酶解增加的抗性的修饰的生长激素多肽的评估
修饰的GH多肽的对蛋白酶解增加的抗性可以用本领域已知的评估蛋白质稳定性、蛋白酶敏感性和抗性和/或GH生物学活性的任何方法加以评估。在一个实施例中,蛋白酶抗性通过将修饰的GH多肽与一或多种蛋白酶保温然后评估与未处理的对照相比的残留生物学活性而测量。修饰的GH多肽可以与在类似的条件下处理的未修饰和/或野生型天然GH相比以确定是否特定的变体保留比未修饰的GH更高的生物学活性。生物学活性可以用本领域已知的任何方法进行评估,如通过测量增加的肌肉质量、抗衰老和增殖活性而评估。
也可以使用蛋白酶抗性的动力学研究来评估修饰的GH多肽。例如,将修饰的GH多肽与一或多种蛋白酶保温并在一系列时间点取样。在每个时间点,失活蛋白酶,然后测试样品的GH生物学活性。修饰的GH可以与未处理的对照和类似处理的未修饰的和/或野生型GH相比以确定修饰的GH多肽的蛋白酶抗性。
本发明提供的修饰的GH多肽展示对蛋白酶的蛋白酶解的增加的抗性,所述蛋白酶包括在例如体液和组织中出现的蛋白酶,如包括但不限于在唾液、血液、血清、肠道、胃、血液、细胞裂解物、细胞及其它中出现的蛋白酶。修饰可以包括对一或多种蛋白酶的抗性,所述蛋白酶包括但不限于胃蛋白酶、胰蛋白酶、胰凝乳蛋白酶、弹性蛋白酶、氨基肽酶、明胶酶B、明胶酶A、α-胰凝乳蛋白酶、羧基肽酶、内切蛋白酶Arg-C、内切蛋白酶Asp-N、内切蛋白酶Glu-C、内切蛋白酶Lys-C和胰蛋白酶、鲁米那胃蛋白酶、微绒毛内肽酶、二肽基肽酶、肠肽酶、水解酶、NS3、弹性蛋白酶、因子Xa、粒酶B、凝血酶、胰蛋白酶、纤溶酶、尿激酶、tPA和PSA.
对蛋白酶解的抗性是指蛋白酶对修饰的多肽的靶氨基酸残基的裂解与由相同的蛋白酶在相同条件下对未修饰的多肽的裂解相比的任何量的降低。本发明提供的修饰的GH多肽展示对蛋白酶解的增加的抗性,例如,展示比未修饰的GH多肽高至少1%、2%、3%、4%、5%、6%、7%、8%、9%、10%、...20%、...30%、...40%、...50%、...60%、...、70%、...80%、...90%、91%、92%、93%、94%、95%、96%、97%、98%,99%或100%的对蛋白酶解的抗性。
提供了对明胶酶B更具抗性的修饰的GH多肽。修饰赋予多肽比由明胶酶B裂解的未修饰的GH多肽更高的对明胶酶B裂解的抗性。在典型地包括Met-Ser-Tyr-Asn或相应序列的序列处或其附近多肽被明胶酶B裂解。裂解可以发生在这一区域内或附近,如位于这一区域的C-末端或N-末端5、10、15或约20个氨基酸之内。如果需要,多肽中的被明胶酶B裂解的精确氨基酸可以凭经验确定。因此,本发明提供了这样的多肽,其在氨基酸序列Met-Ser-Tyr-Asn中的氨基酸或相应氨基酸中被修饰或者在这一序列或相应序列之前或之后的残基中被修饰。修饰的多肽可以在每个残基Met、Ser、Tyr、Asn或其组合处被修饰,或者在这一区域附近的氨基酸处被修饰由此使修饰的多肽比未修饰的多肽对明胶酶B的蛋白酶解更具抗性。修饰的多肽比未修饰的多肽对明胶酶B的蛋白酶解的抗性高至少0.5%、1%、5%、10%、15%、20%、25%、30%、35%、40%、45%、50%、55%、60%、65%、70%、75%、80%、85%、90%、95%、96%、97%、98%、99%或100%。在一个实施方案中,对蛋白酶的抗性可以通过本文描述的任一项测定经验测试。
在一个实施方案中,鉴别的用于增加蛋白酶抗性的修饰的位置包括例如F1、P2、P5、L9、D11、M14、R16、L23、D26、K41、Y42、E56、E65、E66、L73、E74、L81、L87、L101、Y111、D112、D116、E119、L124、M125、P133、R134、K140、D147、D153、L156、L157、K158、L162、F166、R167、K168、D169、D171、D171Q、K172、E174、L177、R178和F191。与未修饰的hGH相比展示增加的蛋白酶抗性的修饰的GH多肽被鉴别,它们包括但不限于F1I、P2A、P5S、L9V、D11N、M14V、R16H、L23I、L23V、D26N、K41Q、Y42H、Y42I、E56Q、E56N、E65Q、E66Q、L73V、E74N、L81V、L87V、L101V、Y111I、D112N、D116Q、E119Q、L124V、M125I、M125V、P133A、R134A、R134H、K140N、D147N、D147Q、D153N、L156I、L157I、K158N、L162I、F166I、R167H、R167Q、K168N、K168Q、D169Q、D171N、D171Q、K172Q、E174Q、E174N、E174H、L177V、L177I、R178Q和F191I。
2.增加的热耐受性
在修饰的GH多肽中,本发明提供了被修饰以通过改善热耐受性而增加稳定性的GH多肽。这类修饰可以包括例如增加的螺旋之间盐水(极性)相互作用和增加的螺旋之间H键相互作用。
a.耐热的修饰的生长激素多肽的性质
在一个实施例中,展示增加的热耐受性的修饰的GH多肽是人类GH多肽。可以使用2D-扫描方法学鉴别GH多肽上导致改善的热耐受性的的氨基酸变化。例如,如本文所述,为增加hGH的热耐受性,第一个条件是与分子的热耐受性相关的氨基酸的性质,如其潜在参与能参与分子稳定化的化学键。第二个前提典型地与那些氨基酸在蛋白质结构上的特异位置。通过置换氨基酸可以在GH多肽序列中进行若干结构修饰以增加GH多肽的构象稳定性同时改善或保持所需的生物学活性(例如,细胞增殖活性)。在举例性的实施方案中,这些修饰产生具有由增加的热耐受性评估的改善的稳定性的修饰的GH多肽。这类修饰的GH多肽与未修饰的和/或野生型天然GH多肽相比展示增加的蛋白质半衰期。这些修饰包括疏水性patches的修饰以增加与溶剂的极性相互作用和增加特定螺旋之间的极性相互作用。
在一个实施方案中,与暴露于20℃-45℃之间的特定热条件下的天然GH的半衰期相比,本发明提供的修饰的GH多肽的体外或体内半衰期(血清稳定性)增加了至少10%、至少20%、至少30%、至少40%、至少50%、至少60%、至少70%、至少80%、至少90%、至少100%、至少150%、至少200%、至少250%、至少300%、至少350%、至少400%、至少450%、至少500%或更多。在一个实施例中,热耐受性是在室温(例如约25℃)评估。在另一个实施例中,热耐受性在哺乳动物体温例如对于人类为约37℃评估。在其它实施方案中,与暴露于20℃-45℃之间的特定热条件下例如在室温(例如约25℃)和/或哺乳动物体温(例如约37℃的人类体温)保温的天然GH的半衰期相比,本发明提供的修饰的GH多肽的体外或体内半衰期(血清稳定性)增加了至少6倍、7倍、8倍、9倍、10倍、20倍、30倍、40倍、50倍、60倍、70倍、80倍、90倍、100倍、200倍、300倍、400倍、500倍、600倍、700倍、800倍、900倍、1000倍或更高。
i.分子内键的产生
在互相面对的螺旋之间产生分子键以增加hGH多肽的整体结构稳定性和改善热耐受性。在一个特定的实施方案中,螺旋A和C通过添加电荷或增加这些螺旋之间的极性相互作用而被稳定化。被选择用作is-HIT的氨基酸和氨基酸位置朝向螺旋互相面对面的区域。选择is-HIT时可以考虑溶剂可及性。
鉴别了is-HIT并产生了用于修饰的hGH多肽的更高的热耐受性的LEAD。在每个is-HIT位置的天然氨基酸可以包括但不限于L6、L9、A13、L15、A17、L20、L23、A24、A105、V110、L113、L114、L117、I121、L124和L128(见表5)。在每一个is-HIT位置的天然氨基酸被增加与其它氨基酸的极性相互作用的残基置换:E、D、K、R、N、Q、S或T。
is-HIT和LEAD可以包括在对热稳定性有贡献的特定区域的修饰。在一个实施方案中,选择用于修饰的区域包括在相应于垂体hGH(SEQ ID NO:1)的位置的区域中的一或多个氨基酸修饰。这些区域中的位置包括例如位置1-12、14-26、29-53、57、58、60-63、67-78、80-84、86、87、89、91、93、95-100、102-113、115-128、131、132、135-139、141、142、144、148-160、162-182、185和187-191。例如,垂体hGH被修饰以在相应于SEQ ID NO:1的位置1-12、14-26、29-53、57、58、60-63、67-78、80-84、86、87、89、91、93、95-100、102-113、115-128、131、132、135-139、141、142、144、148-160、162-182、185和187-191中的任何位置的氨基酸位置中包括一或多个氨基酸修饰。在另一个实施方案中,胎盘hGH被修饰。胎盘hGH(SEQ ID NO:712)与垂体hGH(SEQ ID NO:1)进行序列对比,并且从垂体hGH的位置1-12、14-26、29-53、57、58、60-63、67-78、80-84、86、87、89、91、93、95-100、102-113、115-128、131、132、135-139、141、142、144、148-160、162-182、185和187-191选择一或多个氨基酸位置。序列对比和位置选择可以用任何GH多肽进行,即将其与垂体hGH进行序列对比并选择用于修饰的相应位置。
在一个实施方案中,相应于垂体hGH的位置被选择(is-HIT)并进行氨基酸置换(LEAD)以增加的热耐受性。在一个特定的实施方案中,导致增加的热耐受性的氨基酸置换包括例如L6E、L6D、L6K、L6R、L6N、L6Q、L6S、L6T、L9E、L9D、L9K、L9R、L9N、L9Q、L9S、L9T、A13E、A13D、A13K、A13R、A13N、A13Q、A13S、A13T、L15E、L15D、L15K、L15R、L15N、L15Q、L15S、L15T、A17E、A17D、A17K、A17R、A17N、A17Q、A17S、A17T、L20E、L20D、L20K、L20R、L20N、L20Q、L20S、L20T、L23E、L23D、L23K、L23R、L23N、L23Q、L23S、L23T、A24E、A24D、A24K、A24R、A24N、A24Q、A24S、A24T、A105E、A105D、A105K、A105R、A105N、A105Q、A105S、A105T、V110E、V110D、V110K、V110R、V110N、V110Q、V110S、V110T、L113E、L113D、L113K、L113R、L113N、L113Q、L113S、L113T、L114E、L114D、L114K、L114R、L114N、L114Q、L114S、L114T、L117E、L117D、L117K、L117R、L117N、L117Q、L117S、L117T、I121E、I121D、I121K、I121R、I121N、I121Q、I121S、I121T、L124E、L124D、L124K、L124R、L124N、L124Q、L124S、L124T、L128E、L128D、L128K、L128R、L128N、L128Q、L128S和L128T。在一个实施例中,这类氨基酸置换在垂体hGH(SEQ ID NO:1;参见表6和SEQ ID NO:224-351)中进行。
ii.增加螺旋之间的极性相互作用
hGH多肽的螺旋A和C的疏水区被保护不暴露于溶剂。这些疏水区从螺旋A和C的相互作用产生并且在hGH结构的整体稳定化中起作用。也可以进行螺旋A和C的疏水区中的变化以有利于与溶剂的极性相互作用,由此稳定蛋白质构象。
在一个实施方案中,使用蛋白质进化的2D-扫描方法以向螺旋A和C的疏水区中加入电荷,由此增加热耐受性。热耐受性的增加可以表现为蛋白质体外半衰期增加和/或蛋白质体内半衰期增加。如本文所述,用于设计和产生具有更长半衰期的高度稳定的保持时间更长的蛋白质的方法包括例如i)鉴别蛋白质序列上能参与螺旋A和C的相互作用和疏水区产生的一些或全部可能的靶位点(这些位点在本文中被称为is-HIT);ii)鉴别特异于每个is-HIT的合适的置换氨基酸,以便在置换该特异is-HIT的一或多个原始(如天然)氨基酸时,可以预期置换会增加is-HIT的稳定性,同时保持或改善蛋白质的所需的生物学活性和特异性;iii)在每一个特异性is-HIT靶位置系统性导入特异的置换氨基酸(候选LEAD)以产生含有相应的突变候选先导分子的文库。产生并生产修饰的GH多肽。修饰的GH多肽然后可以在可寻址阵列中进行一个接一个的表型鉴定,从而每个突变体分子最初在仅一个is-HIT位点含有一个氨基酸置换。在特定的实施方案中,在后续轮中也可以产生含有多个已被候选LEAD氨基酸置换的HIT位点的突变体分子(super-LEAD)。
在另外的实施方案中,这些候选LEAD可被例如本文所述的另外的赋予蛋白质稳定性的突变进一步修饰。例如,候选LEAD可被修饰以包括例如增加螺旋之间的相互作用和/或去除蛋白酶敏感位点。
b.耐热的修饰的生长激素多肽的评估
修饰的GH多肽热耐受性可用本领域已知的任何评估蛋白质稳定性、热变性和/或生物学活性的方法进行评估。在一个实施方案中,热耐受性的动力学通过在特定温度例如在20℃和45℃之间测试活性而测量。在一个实施例中,热耐受性在37℃评估。简而言之,将修饰的GH多肽在选定的温度保温并在各时间点取样以评估与未处理对照相比的残留生物学活性。评估可以包括例如GH活性的细胞增殖测定。热耐受性是基于与在类似处理中的未修饰的GH保持活性的能力相比的修饰的GH多肽在特定温度随时间保持生物学活性的能力而评估。
3.Super-LEAD和另外的生长激素修饰
GH修饰还可以包括组合两或多个修饰,例如,两个或多个LEAD可以被组合成一个单一的新分子。增加蛋白酶解抗性的修饰可以与本文提供的或现有技术已知的增加蛋白酶解抗性的其它修饰组合。增加热耐受性的修饰可以与本文提供的或现有技术已知的增加热耐受性的其它修饰组合。增加热耐受性的修饰可以与本文提供的或现有技术已知的增加蛋白酶解抗性的修饰组合。增加热耐受性和/或蛋白酶稳定性的修饰也可以与改变其它功能如生物学活性、受体相互作用的GH修饰、影响翻译后蛋白质修饰的修饰以及本领域的任何其它已知修饰组合。
已知有许多将修饰组合在一个多肽中的技术。例如,AdditiveDirectional Mutagenesis(ADM)可以被用于在一个单突变蛋白质上组装存在于各个LEAD分子上的多个突变,以产生super-LEAD突变蛋白质(参见共同在审美国申请No.10/658,355;U.S.申请公开No.US-2004-0132977-A1和公开的PCT申请WO 2004/022747和WO2004/022593)。ADM是一种重复多步骤方法,其中在产生第一个LEAD突变蛋白质之后的每一步骤,一个新的LEAD突变被加到前一个LEAD突变蛋白质上以产生后续的super-LEAD突变蛋白质。
产生编码新的super-LEAD突变分子集合的核酸分子集合群,在可寻址阵列中一个接一个测试和表型鉴定。Super-LEAD突变分子是含有变化数目和类型的LEAD突变的分子。展示针对特定的被进化特征的改善的适合性的分子被称为super-LEAD。Super-LEAD可以通过本领域技术人员已知的其它方法产生并通过本文的高通量方法测试。为本文目的,super-LEAD典型地具有以希望的量不同于LEAD的改善的活性的针对感兴趣的生物学活性或功能的活性,所述的量例如不同于衍生其的至少一个LEAD突变体至少10%,20%,30%,40%,50%,60%,70%,80%,90%,100%,150%,200%或更多。在其它实施方案中,活性的变化比衍生其的至少一个LEAD分子高至少约2倍、3倍、4倍、5倍、6倍、7倍、8倍、9倍、10倍、20倍、30倍、40倍、50倍、60倍、70倍、80倍、90倍、100倍、200倍、300倍、400倍、500倍、600倍、700倍、800倍、900倍、1000倍或更多倍。与LEAD一样,super-LEAD的活性变化取决于正在被“进化”的活性。希望的改变可以是活性的增加或降低,其取决于感兴趣的功能或性质。
可以用来产生两个或多个突变的组合的另一个方法是使用称作“多重叠引物延伸”的寡核苷酸介导的诱变(参见共同在审美国申请No.10/658,355,美国申请公开No.US-2004-0132977-A1和公开的PCT申请WO 2004/022747和WO 2004/022593)。这一方法可用于合理组合突变的LEAD以形成super-LEAD。这一方法使得可以在已知序列的小蛋白质或蛋白质区域中同时导入几个突变。从编码突变的LEAD蛋白质的DNA序列(基因)设计典型地长度约为70个碱基的重叠寡核苷酸(因为更长的寡核苷酸导致错误增加),以便它们在典型地约为20个碱基的区域上互相重叠。尽管典型地使用约70个碱基产生重叠寡核苷酸,但是所使用的另外的重叠寡核苷酸的长度可以从30个碱基直至约100个碱基。类似地,尽管重叠寡核苷酸的重叠区域典型地是约20个碱基,但是本文所使用的其它重叠区域的长度可以从约5个碱基直至约40个碱基。这些长度寡核苷酸(包括或不包括点突变)在第一步PCR(使用校正聚合酶,例如Pfu DNA聚合酶以避免未预料到的突变)中作为模板和引物以产生少量全长基因。得自第一步PCR的全长基因随后在第二步PCR中被选择性扩增,所述PCR使用侧翼引物,每个引物附有限制酶位点以便于随后的克隆。一个多重叠延伸过程产生全长(多突变)核酸分子,其编码具有多个衍生自LEAD突变蛋白质的突变的候选super-LEAD蛋白质。
E.修饰的生长激素多肽的生产
1.表达系统
人类GH多肽可以通过本领域已知的用于蛋白质生产的任何方法生产,包括将编码hGH的核酸分子导入宿主细胞、宿主动物中和/或在体外从编码hGH的核酸分子进行表达。表达宿主包括E.coli、酵母、昆虫细胞、哺乳动物细胞包括人类细胞系以及转基因动物。表达宿主可以在蛋白质生产水平和表达的蛋白质上存在的翻译后修饰类型方面不同。表达宿主的选择可以根据这些和其它因素如医药报批和安全性考虑、生产成本和纯化的需要和方法等进行。
在真核宿主中的表达可以包括在酵母如酿酒酵母和巴斯德毕赤氏酵母、昆虫细胞如果蝇细胞和lepidopteran细胞、植物和植物细胞如烟草、玉米、水稻、藻类和lemna中的表达。用于表达的真核细胞还包括哺乳动物细胞系如中国仓鼠卵巢(CHO)细胞。真核表达宿主还包括在转基因动物中生产,例如包括在牛奶和鸡蛋中生产。
有许多表达载体可用来表达hGH。表达载体的选择由宿主表达系统的选择而影响。这种选择在本领域技术人员的技能范畴。通常,表达载体可以包括转录启动子和任选地增强子、翻译信号以及转录和翻译终止信号。用于稳定转化的表达载典型地具有选择标记,使得可以选择和维持转化的细胞。在某些情况下,可以使用复制起点来扩增载体的拷贝数。
a.原核表达
原核生物特别是大肠杆菌提供了生产大量hGH的系统(参见例如,Platis et al.Protein Exp.Purif.31(2):222-30(2003);Khalizzadeh etal.J.Ind.Microbiol.Biotechnol.31(2):63-69(2004))。大肠杆菌的转化是本领域技术人员熟知的简便快速的技术。用于大肠杆菌的表达载体可以含有可用于诱导高水平蛋白质表达和表达显示对宿主细胞的某些毒性的蛋白质的诱导型启动子。诱导型启动子的例子包括lac启动子、trp启动子、杂合tac启动子、T7和SP6 RNA启动子以及温度调节的λPL启动子。
人类GH可以在大肠杆菌的细胞质环境中表达。细胞质是还原性环境并且对于一些分子这可以导致不溶性包涵体的形成。还原剂如二硫苏糖醇和β-巯基乙醇和变性剂(例如胍-HCl和脲)可以用于重溶蛋白质。一种替代方法是在细菌的周质空间表达hGH,周质空间提供了氧化环境和陪伴蛋白样和二硫化物异构酶并且可以导致形成可溶蛋白质。典型地,前导序列与感兴趣的蛋白质融合,将蛋白质导向周质。前导序列随后被周质中的信号肽酶除去。周质靶向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,周质表达使得表达的蛋白质泄漏进培养基中。蛋白质的分泌使得可以从培养物上清快速简便地纯化。未被分泌的蛋白质可以通过渗透压裂解从周质获得。与细胞质表达类似,在一些情况中蛋白质可以变得不溶,可以使用变性剂和还原剂以促进溶解和再折叠。诱导和生长的温度也可以影响表达水平和溶解性。典型地,使用25℃至37℃之间的温度。也可以使用突变来增加表达的蛋白质的溶解性。典型地,细菌产生非糖基化蛋白质。因此,如果蛋白质功能需要糖基化,可以在从宿主细胞纯化后体外添加糖基化。
b.酵母
酵母如酿酒酵母(Saccharomyces cerevisae)、粟酒裂殖酵母(Schizosaccharomyces pombe)、解脂耶氏酵母(Yarrowialipolytica)、乳酸克鲁维酵母(Kluyveromyces lactis)和巴斯德毕赤氏酵母(Pichia pastoris)是hGH的有用表达宿主(参见例如,Skokoet al.Biotechnol.Appl.Biochem.38(Pt3):257-265(2003))。酵母可以用附加型复制载体转化或通过同源重组进行的稳定染色体整合而转化。典型地,使用诱导型启动子调节基因表达。这种启动子的例子包括GAL1、GAL7和GAL5和金属硫蛋白启动子如CUP1。表达载体通常包括选择标记如LEU2、TRP1、HIS3和URA3以用于选择和保持转化的DNA。在酵母中表达的蛋白质通常是可溶。与陪伴蛋白如Bip和蛋白质二硫化物异构酶的共表达能够改善表达水平和溶解性。另外,在酵母中表达的蛋白质可以被导向分泌,分泌使用分泌信号肽融合体如来自酿酒酵母的酵母交配型α因子分泌信号以及与酵母细胞表面蛋白如Aga2p交配粘附受体或Arxula adeninivorans葡糖淀粉酶的融合体。蛋白酶裂解位点如Kex-2蛋白酶裂解位点可以被工程化引入以便多肽从分泌途径出来后从多肽中除去融合的序列。酵母还能在Asn-X-Ser/Thr基序糖基化。
c.昆虫和昆虫细胞
昆虫和昆虫细胞、特别是使用杆状病毒表达的昆虫和昆虫细胞可用于表达生长激素如hGH(参见例如,Muneta et al.J.Vet.Med.Sci.65(2):219-23(2003)。昆虫细胞和昆虫幼虫、包括在血淋巴中的表达能表达高水平的蛋白质并且能进行高等真核生物使用的大多数翻译后修饰。杆状病毒具有有限的宿主范围其改善了安全性并降低了真核表达的法规报批考虑。典型的表达载体使用用于高水平表达的启动子如杆状病毒多角体蛋白启动子。常用的杆状病毒系统包括杆状病毒如Autographa californica核多角体病毒(AcNPV)和bombyx mori核多角体病毒(BmNPV)以及昆虫细胞系如衍生自Spodopterafrugiperda的Sf9、Pseudaletia unipuncta(A7S)和Danaus plexippus(DpNl)。为了高水平病毒,待表达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并可用于将表达的蛋白质分泌进培养基。另外,细胞系Pseudaletia unipuncta(A7S)和Danaus plexippus(DpNl)产生具有类似于哺乳动物细胞系统的糖基化模式的蛋白质。
昆虫细胞中的另一种表达系统是利用稳定转化的细胞。细胞系如Schnieder 2(S2)和Kc细胞(Drosophila melanogaster)和C7细胞(Aedes albopictus)可用于表达。果蝇金属硫蛋白启动子在用镉或铜进行的重金属诱导存在下可以用于诱导高水平表达。表达载体典型地用选择标记如新霉素和潮霉素保持。
d.哺乳动物细胞
哺乳动物表达系统可以用于表达GH多肽。表达构建体可以通过病毒感染如腺病毒或通过直接DNA转移如脂质体、磷酸钙、DEAE-葡聚糖以及通过物理手段如电穿孔和显微注射而被转移进哺乳动物细胞。用于哺乳动物细胞的表达载体典型地包括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。载体经常包括用于高水平表达的转录启动子-增强子如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有活性。组织和细胞类型启动子和增强子区域也可以用于表达。举例性的启动子/增强子区域包括但不限于来自基因如弹性蛋白酶I、胰岛素、免疫球蛋白、小鼠乳腺肿瘤病毒、白蛋白、甲胎蛋白、α1抗胰蛋白酶、β-珠蛋白、髓鞘碱性蛋白、肌球蛋白轻链-2和促性腺激素释放激素基因控制的那些区域。选择标记可用于选择和保持具有表达构建体的细胞。选择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、次黄嘌呤-鸟嘌呤磷酸核糖基转移酶、氨基糖苷磷酸转移酶、二氢叶酸还原酶和胸苷激酶。具有细胞表面信号传导分子如TCR-ζ和FcεRI-γ的融合体可以指导活性状态的蛋白质在细胞表面的表达。
有许多细胞系可用于哺乳动物表达,包括小鼠、大鼠、人、猴、鸡和仓鼠细胞。举例性的细胞系包括但不限于CHO,Balb/3T3,HeLa,MT2,小鼠NS0(非分泌型)和其它骨髓瘤细胞系、杂交瘤和异源杂交瘤(heterohybridoma)细胞系、淋巴细胞、成纤维细胞、Sp2/0、COS、NIH3T3、HEK293、293S、2B8和HKB细胞适应于无血清培养基的细胞系也是可获得,其促进分泌的蛋白质从细胞培养基中纯化。一个例子是无血清EBNA-1细胞系(Pham et al.,Biotechnol.Bioeng.84:332-42(2003))。
e.植物
转基因植物细胞和植物可以用于表达hGH。典型地用直接DNA转移如微粒轰击和PEG介导的转移进原生质体和用农杆菌介导的转化将表达构建体转移进植物。表达载体可以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通常分为双子叶宿主如Arabidopsis和烟草以及单子叶宿主如玉米和水稻。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和UBQ3启动子。经常使用选择标记如潮霉素、磷酸甘露糖异构酶和新霉素磷酸转移酶来促进转化的细胞的选择和保持。转化的植物细胞可以在培养物如细胞、聚集体(愈伤组织)中保持或再生成完整植物。转基因植物细胞也可包括生产蛋白质的工程藻类(参见例如,Mayfield et al.(2003)PNAS 100:438-442)。因为植物具有与哺乳动物细胞不同的糖基化模式,因此这可以影响在这些宿主中生产hGH的选择。
2.纯化
从宿主细胞纯化GH多肽的方法取决于所选的宿主细胞和表达系统。对于被分泌的分子,蛋白质通常在除去细胞后从培养基中纯化。对于胞内表达,细胞可以被裂解并从提取物中纯化蛋白质。当使用转基因生物如转基因植物和动物进行表达时,可以使用组织或器官作为起始材料以制备裂解的细胞提取物。另外,转基因动物生产可包括在奶或蛋中生产多肽,它们可以被收集并且如果需要可以用本领域标准方法进一步提取蛋白质和进一步纯化。
生长激素可以用本领域已知的标准蛋白质纯化技术纯化,这些技术包括但不限于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。亲和纯化技术也可以被用来改善效率和制备物的纯度。例如,抗体、受体和结合GH的其它分子可以用于亲和纯化。表达载体还可以工程化以向蛋白质添加亲和标记如myc表位、GST融合体或His6并且分别用myc抗体、谷胱甘肽树脂和Ni树脂亲和纯化。纯度可以用本领域已知的任何方法评价,包括凝胶电泳和染色以及分光光度技术。
3.融合蛋白
还提供了含有靶向剂和修饰的GH多肽的融合蛋白。提供了配制用于经合适途径给药的含有这种融合蛋白的药物组合物。融合蛋白是通过以任何顺序连接修饰的GH和一种物质如抗体或其片段、生长因子、受体、配体和其它将突变蛋白质导向靶细胞或组织的物质。连接可以直接进行或经接头间接实现。融合蛋白可以重组产生或经化学键化学产生,如经异双功能试剂或硫醇键或其它这类键。融合蛋白可以含有另外的成分如大肠杆菌麦芽糖结合蛋白(MBP)以助于蛋白质被细胞摄入(参见例如,国际PCT申请No.WO01/32711)。
4.多肽修饰
修饰的GH多肽可以作为裸多肽链或作为复合物而制备。对于某些应用,需要制备不含翻译后或其它化学修饰的“裸”形式的修饰的GH。裸多肽链可以在不翻译后修饰GH的合适宿主中制备。多肽也可以在体外系统中使用化学肽合成进行制备。对于其它应用,特定的修饰可以是希望的,包括聚乙二醇化、白蛋白化、糖基化、磷酸化或其它已知的修饰。这些修饰可以体外进行,或者例如通过在产生这些修饰的合适宿主中生产修饰的GH而实现修饰。
5.核苷酸序列
本发明还提供了与启动子如用于在哺乳动物细胞中表达的诱导型启动子可操纵地连接的编码本发明的修饰的GH多肽或融合蛋白的核酸分子。这类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启动子如应答PV E2蛋白的PBV p89启动子;以及由HIV或PV或癌基因激活的其它启动子。
本发明提供的修饰的GH多肽也可以含在基因转移载体中被输送到细胞。所述转移载体还可以编码额外的用于治疗生长激素介导的疾病或状况的另外的治疗剂,用于治疗生长缺陷、HIV感染和其它用修饰的GH给药的疾病。编码修饰的GH多肽的转移载体可以通过将核酸给予对象而全身性使用。例如,转移载体可以是病毒载体如腺病毒载体。编码GH的载体还可以掺入干细胞中并且干细胞可以通过在治疗部位移植(transplanting or engrafting)干细胞而被给予对象。
F.评价修饰的生长激素多肽活性
GH生物学活性可以在体外和/或体内评价。例如,GH变体可以与未修饰的和/或野生型GH比较而在体内评价。GH变体还可以在体内被测试以确定生物学活性、稳定性(例如半衰期)和治疗效果。体内测定包括在动物模型中的GH测定以及对人类给药。
1.体外测定
体外测定包括蛋白质稳定性和生物学活性的测定。稳定性测定可以包括确定蛋白酶抗性、热稳定性和本领域已知的其它蛋白质结构和构象分析。生物学活性测定可以包括GH与其受体的相互作用的测量以及确定GH变体对GH细胞途径的作用的基于细胞的测定。生物学测定的一个例子是基于细胞的增殖测定。简而言之,测试修饰的GH多肽在细胞系如Nb2-11C细胞(大鼠淋巴瘤细胞系)中刺激细胞增殖的能力。细胞用未修饰的GH或修饰的GH多肽处理。保温后,细胞增殖例如通过细胞计数和/或通过测量存活细胞数(例如用存活染色)而测量。与未修饰的和/或野生型GH多肽的“效力(potency)”相比的“效力”可以通过测量GH的浓度及其活性(即细胞增殖活性)而计算。
2.非人动物模型
非人动物模型是评价生长激素变体的活性和稳定性的有用工具。例如,非人动物可以用作疾病或状况的模型。非人动物可以用诱导疾病和/或表型的物质注射,然后给予生长激素变体以监测对疾病进程的效果。遗传模型也是有用的。通过过表达、低表达或敲除一或多个基因可以产生模拟疾病或状况的动物如小鼠。这类动物可以通过本领域熟知的转基因动物生产技术或用天然发生的或诱导的突变体株系产生。有用的与生长迟缓相关的疾病的非人动物模型包括但不限于矮小动物,包括生长激素缺陷的Lewis株系侏儒大鼠(dwarf rat)(dw/dw)、自发侏儒大鼠(SDR);生长激素缺陷侏儒小鼠(lit/lit或“little mice”)、Ames侏儒小鼠(Prop11df)、Bayer侏儒小鼠、Snell’s侏儒小鼠(Pitldw);猪、长卷毛狗和Brahmin牛的侏儒株系;生长激素受体敲除小鼠(GHR-/-);垂体摘除动物如大鼠;对生长激素作用有抗性的动物物种和株系如豚鼠和Laron小鼠。
这些非人动物模型可以用于监测生长激素变体与野生型生长激素相比的生物学活性。最常见类型的体内GH生物测定是垂体摘除大鼠中的体重增重(Evans et al.Endocrinology 22:483-492(1938);Marxet al.Endocrinology 30:1-10(1942);and Greesbeck et al.Endocrinology120:2582-2590(1987))或侏儒“小”鼠中的体重增重(Bellini et al.Endocrinology 132:2051-2055(1993))。简而言之,侏儒小鼠可以通过经微型渗透泵连续皮下给予突变体或野生型GH而处理。在处理方案后,可以测量指示GH生物学活性的变量,包括但不限于体重增重、身体组成(水、脂肪、与体重相关的蛋白质)的变化、IGF-I或IGFBP-3产生、生长激素受体mRNA水平(例如由RT-PCR测得)、空腹血脂谱、血糖和胰岛素水平。
另一种常见类型的GH活性体内生物测定是在垂体摘除大鼠上的tibial plate assay(Greespan et al.Endocrinology 45:455-463(1949)).简而言之,在性成熟前雄性大鼠中进行垂体摘除术。选择在2周期间每周增重小于2克的动物用于GH输注。微型渗透泵用修饰的GH多肽充满并置于腹部皮下。测量给药之前和给药过程中的食物和水摄入以及体重。对动物经主动脉穿刺放血,经RIA或ELISA分析血清以评价激素的血清浓度。通过在立体显微镜下分析银染的胫骨而确定骺宽(Epiphyseal width),生长速率可以通过用入射光荧光显微镜术测量甲基丙烯酸酯包埋的胫骨切片中的累积纵向骨生长而评价。
在鉴别和测试用于治疗特征在于GH调节和活性改变的其它疾病和病症的GH变体中有用的其它合适的非人动物模型包括但不限于SIV(猿猴免疫缺陷病毒)感染的AIDS消瘦猕猴模型;囊性纤维化的cftr-/-小鼠模型;克罗恩氏病的mdrla-/-或SAMP1/Yit小鼠模型;慢性肾病的肾切除小鼠或大鼠模型。
动物模型可进一步用于监测生长激素变体的稳定性、半衰期和清除。这种测定可以用于比较生长激素变体和计算剂量和剂量方案以用于进一步的非人动物和人试验。例如,修饰的生长激素可以被注射进小鼠的尾静脉。在注射后各时间点(如之后的分钟、小时和天)取血样,然后可以例如通过ELISA或放射免疫测定在特异时间点监测包括但不限于血清或血浆的身体样品中的生长激素变体的水平。
3.临床测定
有许多测定可用来评价临床用的生长激素的生物学活性。这类测定包括评价受体结合、受体活化、体内蛋白质稳定性和半衰期以及表型测定。举例性的测定包括但不限于适合临床使用的体外GH生物测定如放射受体测定(Tsushima et al.,J.Clin.Endocrinol.Metab.37:344-337(1973);and Lesniak et al.Nature 241:20-22(1973))、受体调节测定(Rosenfeld et al.J.Clin.Endocrinol.Metab.50:62-69(1980))以及使用Nb2细胞系的细胞增殖生物测定(Tanaka et al.J.Clin.Endocrinol.Metab.51:1058-1063(1980))。其它技术可以包括使用表达hGH受体的小鼠pro-B细胞淋巴细胞系以建立用于hGH的生物测定系统,测量对象血清中的水平(Ishikawa et al.Journal of Clinical Endocrinology&Metabolism 85(11):4274-4279(2000))。
表型测定和评价GH治疗的治疗效果的测定包括评价GH血液水平(例如在给药前和给药后各时间点测量血清GH和垂体前叶激素,所述给药后各时间点包括第一次给药后、最后一次给药后立即、以及两者之间的时间点,针对体重指数(body mass index,BMI)校正),对GH治疗的表型应答包括与用未修饰的和/或野生型GH或安慰剂处理的对象相比症状随时间的减轻。例如,在刺激生长的治疗中,测定可以包括生长速率测量和IGF-I、IGFBP-3和GHRH血清水平的测量。空腹血脂谱、血糖和胰岛素也可以用放射免疫测定来测量。
G.配制/给药
含有本文产生的变体、包括hGH变体多肽、hGH融合蛋白或编码核酸分子的药物组合物可以任何常规方式配制,即,将选择量的多肽与一或多种生理学可接受的载体或赋形剂混合。载体或赋形剂的选择属于给药专业的技能并且可以依赖于许多参数。这些参数包括例如给药模式(即全身性、口服、肺、局部(local)、局部(topical)或任何其它模式)和被治疗的状况。本发明提供的药物组合物可以配制用于单剂量(直接)给药或者用于稀释或其它修改。配制品中化合物的浓度是给药时有效输送进行有效目的治疗的量。典型地,组合物被配制为单剂量给药。为了配制组合物,将化合物或其混合物的重量份以有效减轻或改善被治疗的状况的浓度溶解、悬浮、分散或以其它方式混合在选定的载体中。适于本发明化合物的给药的药物学载体包括本领域技术人员已知的适于特定给药模式的任何这类载体。
多肽可以作为唯一药物活性成分而配制在组合物中,或者可以与其它活性成分组合。多肽可以被定向输送,如与靶向剂如抗体缀合。脂质体悬浮液、包括组织靶向脂质体也可以适于作为药物学可接受的载体。它们可以根据本领域技术人员已知的方法制备。例如,可以如美国专利4,522,811所述制备脂质体配制品。脂质体输送也可以包括缓释配制品,包括药物基质如胶原蛋白凝胶好用纤连蛋白修饰的脂质体(参见例如,Weiner et al.J.Pharm.Sci.74(9):922-5(1985))。
活性化合物以足以在被治疗的对象中引起治疗有用效果并且不存在不希望的副作用的量包括在药物学可接受的载体中。治疗有效浓度可以通过在已知的体外和体内系统如本文提供的测定中测试化合物而经验性确定。活性化合物可以液体、半液体或固体形式以任何合适的途径给药,例如,口服、鼻、肺、肠胃外、静脉内、皮内、皮下或局部,并以适于每种给药途径的方式配制。
可以配制修饰的hGH及生理学可接受的盐和溶剂化物以用于吸入给药(经口或经鼻)、口服、肺、透皮、肠道外或直肠给药。对于吸入给药,修饰的hGH可以以气溶胶喷雾形式输送,气溶胶喷雾来自加压包装或使用合适的推进剂的喷雾器,所述推进剂例如是二氯二氟甲烷、三氯氟甲烷、二氯四氟乙烷、二氧化碳或其它合适的气体。在加压气溶胶情况中,剂量单位可以通过提供一个阀而输送计量的量而确定。可以配制含有治疗化合物和合适的粉基如乳糖或淀粉的粉末混合物的例如明胶胶囊或药筒以用于吸入器或吹入器中。
对于肺给药至肺部,修饰的hGH可以以气溶胶喷雾形式输送,该气溶胶来自使用合适推进剂的喷雾器、turbonebulizer或微处理器控制的计量剂量口腔吸入器。通常颗粒大小较小,如在0.5至5微米范围内。当药物组合物配制用于肺部给药时,典型地不使用去污剂表面活性剂。肺部药物输送是一种有前景的非侵入性全身性给药方法。肺代表了一种有吸引力的药物输送途径,这主要是由于其有用于吸收的高表面积、薄的肺泡上皮、广泛的血管化、不存在肝首过代谢(hepatic first-pass metabolism)以及相对较低的代谢活性。
修饰的hGH多肽可以作为depot preparation配制。这种长效作用配制品可以通过植入(例如皮下或肌肉内)或通过肌肉内注射而给药。因此,例如,治疗化合物可以与合适的聚合或输水材料(例如,作为在可接受的油中的乳液)、离子交换树脂或作为难溶衍生物例如难溶盐而配制。
修饰的hGH可以配制用于经注射而肠胃外给药(例如经推注或连续输注)。用于注射的配制品可以以添加防腐剂的单位剂型存在(例如在安瓿中或在多剂容器中)。组合物可以呈如下形式:在油或水性载体中的悬浮液、溶液或乳液并且可以含有配制剂如悬浮、稳定和/或分散剂。或者,活性成分可以是粉末冻干形式,在使用前用合适的载体例如无菌无热原水配制。
活性成分可以配制用于局部(local)或局部(topical)应用,如用于以凝胶、霜和洗剂形式局部应用于皮肤(透皮)和粘膜如在眼中,以及用于应用于眼或用于脑池内或脊柱内应用。这类溶液、特别是要用于眼科用途的溶液可以用合适的盐配制为0.01%-10%等渗溶液且pH约为5-7。化合物可以配制为气溶胶以用于局部应用如通过吸入应用(参见例如美国专利4,044,126,4,414,209和4,364,923,它们描述了用于输送类固醇以用于治疗炎症特别是哮喘的气溶胶)。
药物组合物中的活性化合物的浓度取决于活性化合物的吸收、失活和排泄率、剂量计划、给药量以及本领域技术人员已知的其它因素。如本文所述,可以使用本领域已知的用于未修饰的hGH给药的剂量并比较修饰的hGH与未修饰的和/或天然hGH相比的性质和活性(例如稳定性和生物学活性)而经验性确定剂量。
如果需要,药物组合物可以存在于一个包装中、一个试剂盒或分配装置中,所述包装、试剂盒或分配装置可以含有一或多个含活性成分的单位剂型。所述包装例如含有金属或塑料箔,泡罩包装(blister pack)。所述包装或或分配装置可以附有给药说明。含有活性成分的药物组合物可以被包装成含有包装材料、本文提供的药剂和指示所述药剂治疗何种疾病的标签的制品。
在修饰的GH多肽中,本发明提供了增加稳定性的修饰的GH多肽以适于口服输送条件。口服输送可以包括给药于口腔和/或胃肠道。这类修饰包括在一或多种条件下的增加的蛋白质半衰期,所述条件如暴露于唾液、暴露于胃肠道中的蛋白酶、暴露于特定的pH条件如胃的低pH和/或肠中的pH条件。修饰可包括在胃部低pH中对一或多种蛋白酶包括明胶酶A和明胶酶B的抗性。修饰还可包括增加对潜在的变性或构象改变条件的整体稳定性,如热耐受性以及对混合和充气(例如咀嚼)的耐受性。
被修饰为适于口服输送的生长激素多肽可以用本文描述的任一方法制备。例如,用于蛋白质合理进化的2D-和3D-扫描诱变方法(参见共同在审的美国申请系列号10/685,355和美国专利公开No.US-2004-0132977-A1以及公开的国际申请WO 2004/022593 and WO2004/022747)可以用于制备修饰的GH多肽。适于口服输送的GH的修饰可以包括除去GH中的蛋白酶解消化位点和/或增加GH结构的整体稳定性。与未修饰的和/或野生型天然GH相比,这类修饰的GH展示在口服输送的一或多种条件下的增加的蛋白质半衰期。例如,修饰的GH可以具有在口腔、喉(例如通过粘膜层)、胃肠道或全身性的增加的蛋白质半衰期和/或生物利用度。
在一个实施方案中,当与暴露于口服输送的一或多种条件下的天然GH的半衰期相比时,本发明提供的修饰的GH的体外或体内半衰期(血清稳定性)增加了至少10%、至少20%、至少30%、至少40%、至少50%、至少60%、至少70%、至少80%、至少90%、至少100%、至少150%、至少200%、至少250%、至少300%、至少350%、至少400%、至少450%、至少500%或更多。在其它的实施方案中,当与暴露于口服输送的一或多种条件下的天然GH的半衰期相比时,本发明提供的修饰的GH的体外或体内半衰期(血清稳定性)增加了至少6倍、7倍、8倍、9倍、10倍、20倍、30倍、40倍、50倍、60倍、70倍、80倍、90倍、100倍、200倍、300倍、400倍、500倍、600倍、700倍、800倍、900倍、1000倍或更多。
在一个实施例中,修饰的GH的半衰期由在一或多种蛋白酶存在下增加的半衰期评价。修饰的GH可以与一或多种蛋白酶混和,然后在合适的反应时间后评价生物学活性和/或蛋白质结构。半衰期的评价也可以包括暴露于增加的温度如对象的体温;暴露于胃液和/或模拟的胃液;暴露于特定的pH条件和/或两种或多种条件的组合。在暴露于一或多种条件之后,生物学活性和/或GH结构的评价可用于评价与合适对照(即未修饰的和/或野生型GH蛋白质)比较的修饰的GH的半衰期。
修饰的hGH多肽可以配制为用于口服给药,如配制为片剂、胶囊剂、液体或其它用于口服给药的合适载体。用于口服输送的含有修饰的GH的药物组合物的制备可以包括用本领域已知的和本文描述的口服配方配制修饰的GH。配制的组合物无需添加蛋白酶抑制剂和/或其它在暴露于蛋白酶、pH和其它口服输送条件时稳定未修饰的和野生型GH所需的成分。例如,这类组合物在不存在化合物如放线酰胺素或表放线酰胺素(epiactinonin)及其衍生物、Bowman-Birk抑制剂及其缀合物、抑酶肽和卡莫他特时展示稳定性。
另外,因为本发明提供的修饰的GH展示增加的蛋白质稳定性,所以在给予所述药物组合物时比给予其未修饰的对应物时更具灵活性。典型地,口服摄入的GH在早饭前给药(即在激活消化酶之前)。本文的修饰的GH展示对消化酶的蛋白酶抗性并能提供在每日的其它时间和在消化酶存在或有活性的条件下给予含有修饰的GH的药物组合物的能力。
为了口服给药,药物组合物可以采取例如片剂或胶囊剂的形式,这些形式使用传统手段用药物学可接受的赋形剂如粘合剂(例如预胶化玉米淀粉、聚乙烯吡咯烷酮或羟丙基甲基纤维素);填充剂(例如乳糖、微晶纤维素或磷酸氢钙);润滑剂(例如硬脂酸镁、滑石粉或二氧化硅);崩解剂(例如马铃薯淀粉或淀粉乙醇酸钠);或润湿剂(例如月桂基硫酸钠)制备。片剂可以用本领域熟知的方法包衣。片剂可以作为补剂摄入以增加肌肉质量。用于口服给药的液体制剂可以采用例如溶液、糖浆或悬浮液形式,或者它们可以呈干燥产物形式并且在使用前用水或其它合适的载体配制。这类液体制剂可以用药物学可接受添加剂经传统手段制备,所述添加剂如悬浮剂(例如山梨糖醇糖浆、纤维素衍生物或氢化食用脂肪);乳化剂(例如卵磷脂或阿拉伯胶);非水载体(例如杏仁油、油状酯、乙醇或分级植物油);和防腐剂(例如对羟基苯甲酸甲酯或丙酯或山梨酸)。如果合适,制剂还可含有缓冲盐、香味剂、着色剂和/或甜味剂。
口服给药制剂可以配制成控制释放或缓释或在穿过胃后释放或在小肠中释放活性化合物。为了口服给药,组合物可采取片剂、胶囊剂、液体、锭剂和其它适合口服给药的形式。适合口服给药的配制品包括锭剂和将药物组合物输送至口腔粘膜、喉和/或胃肠道的其它配制品。锭剂可以用合适的成分配制,所述成分包括赋形剂例如无水结晶麦芽糖和硬脂酸镁。如所述,本文的修饰的GH展示对血液或肠道蛋白酶的抗性并可以不另加蛋白酶抑制剂或其它保护性化合物而配制。用于口服给药的制剂也可以包括与一或多种额外成分配制的抗蛋白酶解的修饰的GH,所述一或多种成分也赋予在其它条件如特定pH条件中的蛋白酶抗性或稳定性。
本发明还提供了编码hGH多肽的核酸分子和编码其的表达载体的药物组合物,它们适于基因治疗。代替输送蛋白质,核酸分子可以被体内给药(例如全身性或其它途径给药)或离体(ex vivo)给药如取出细胞如淋巴细胞、在其中导入核酸并重新导入宿主或相容受体。
人类GH多肽可以通过表达核酸分子而被输送至细胞和组织。人类GH多肽可以作为编码hGH多肽的核酸分子给药,包括离体技术和直接体内表达。
核酸可以通过本领域技术人员已知的任何方法被输送至细胞和组织。分离的核酸序列可以掺入载体中用于进一步操作。如本文所用,载体(或质粒)是指用于将异源DNA导入细胞进行表达或复制的独立的元件。这类载体的选择和使用属于技术人员的技能范畴。
通过表达编码核酸分子而给予hGH多肽的方法包括给予重组载体。载体可以被设计为保持成附加型,例如通过加入复制起点实现,或者可以被设计为整合进细胞的染色体中。人类GH多肽也可以用于使用非病毒载体的离体基因表达治疗。例如,可以工程化细胞以表达hGH多肽,如通过将编码hGH多肽的核酸整合进基因组位置,可操纵地与调节序列连接或者其被置于可操纵地连接于基因组位置中的调节序列。这类细胞随后可以局部或全身性地给予对象如需要治疗的患者。
病毒载体包括例如腺病毒、疱疹病毒、逆转录病毒,也可以采用其它设计用于基因治疗的载体。载体可以保持为附加型或可以整合进被治疗对象的染色体中。HGH多肽可以由被给予需要治疗的对象的病毒表达。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、慢病毒和如上所述的其它病毒。例如,腺病毒表达技术是本领域熟知的并且腺病毒生产和给药方法也是熟知的。腺病毒血清型可例如从美国典型培养物保藏中心(ATCC,Rockville,MD)获得。腺病毒可以离体应用。例如,从需要治疗的患者分离细胞,并用表达hGH多肽的腺病毒载体转导。在合适的培养时间喉,将转导的细胞局部和/或全身性给予对象。或者,分离表达hGH多肽的腺病毒颗粒并配制在药物学可接受载体中以输送治疗有效量,从而预防、治疗或改善对象的疾病或状况。典型地,腺病毒颗粒以从每kg对象体重1个颗粒直至1014个颗粒的剂量范围输送,通常在每kg对象体重106或108个颗粒至1012颗粒之间。在某些情况下,希望给核酸源提供一种靶向细胞的物质,如特异于细胞表面膜蛋白或靶细胞的抗体,或针对靶细胞上的受体的配体。
核酸分子可以被导入人工染色体和其它非病毒载体中。人工染色体如ACES(see,Lindenbaum et al.Nucleic Acids Res.32(21):e172(2004))可以被改造以编码和表达同种型。简而言之,哺乳动物人工染色体(MAC)提供了将大有效载荷的遗传信息以自主复制、非整合形式导入细胞中的手段。在MAC中独特的是基于哺乳动物微卫星DNA的人工染色体表达(ACE)可以在不同物种的细胞系中从头可再现地产生,并可以容易地才宿主细胞染色体纯化。纯化的哺乳动物ACE随后可以被重新导入各种受体细胞系中,在此它们使用ACE系统无需选择压力即能稳定保持长时间。使用这一方法,已在LMTK(-)和CHO细胞中实现了一或两个基因靶的特异加载。
另一种方法是在酵母中的两步基因置换技术,这一技术从以下材料出发:克隆在酵母人工染色体(YAC)中的完整腺病毒基因组(Ad2;Ketner et al.Proc.Natl.Acad.Sci.USA 91:6186-6190(1994))和含有靶向YAC克隆中的特异区域的腺病毒序列的质粒、用于感兴趣基因的表达盒和阳性和阴性选择标记。
编码修饰的GH多肽的核酸可以包囊在载体如脂质体中,或者导入细胞如细菌细胞、特别是减毒细菌,或者导入病毒载体中。例如,当采用脂质体时,与胞吞相关的细胞表面膜蛋白结合的蛋白质可以用于靶向和/或用于促进摄入,例如对于特定细胞类型有向性的衣壳蛋白或其片段、针对在循环过程中经历内在化的蛋白质的抗体、以及靶向细胞内位置并增强细胞内半衰期的蛋白质。
对于离体和体内方法,编码hGH多肽的核酸分子被导入来自合适供体或被治疗对象的细胞中。用于治疗目的的可以被导入核酸的细胞包括例如,适于被治疗疾病或状况的任何希望的可获得的细胞类型,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、胎肝和其它来源的干细胞。
为了离体治疗,从与待治疗对象相容的供体或从待治疗的对象取细胞,将核酸导入这些分离的细胞,并将这些修饰的细胞给予对象。治疗包括直接给药,例如包囊在多孔膜中,植入患者中(参见例如美国专利4,892,538和5,283,187)。适于将核酸转移进体外哺乳动物细胞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法。可以使用DNA输送方法体内表达hGH多肽。这类方法包括核酸的脂质体输送和裸DNA输送,包括例如使用电穿孔、超声和磷酸钙输送的局部和系统性输送。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。
hGH多肽的体内表达可以与另外的分子的表达连在一起。例如,hGH多肽的表达可以与细胞毒性产物的表达连在一起,所述细胞毒性产物如在工程病毒中或在细胞毒性病毒中表达。这类病毒可以被定向至作为治疗作用的靶的特定细胞类型。表达的hGH多肽可以用于增强病毒的细胞毒性。
hGH多肽的体内表达可以包括将编码hGH多肽的核酸分子可操作地与特异的调节序列如细胞特异性或组织特异性启动子连接。人类GH多肽也可以从特异地感染靶细胞类型和/或组织和/或在靶细胞类型和/或组织中复制的载体表达。诱导型启动子可被用于选择性调节hGH多肽表达。
裸核酸形式的或在载体、人工染色体、脂质体或其它载体中的核酸分子可以通过全身性给药、局部(topical)、局部(local)和其它给药途径给予对象。当全身性于体内时,核酸分子或含有核酸分子的载体可以被定向至细胞。
给药也可以是直接的,如给予典型地靶向细胞或组织的载体或细胞。例如,肿瘤细胞和增殖可以是用于体内表达hGH多肽的靶细胞。用于体内表达hGH多肽的细胞也包括患者自体细胞。这些细胞可取自患者,导入用于表达hGH多肽的核酸,然后例如通过注射或移植而给予患者。
本文提供的多核苷酸和表达载体可以用任何合适的方法制备。还提供了包含上述核酸分子的核酸载体,所述核酸分子包括包含编码SEQ ID NO:2-69、75、76、85-107、111、112、115、116、119、120、123-154、164-165、176-216、222和223任一序列的多肽或其功能片段的核苷酸序列的核酸分子。还提供了包含上述核酸分子的核酸载体和含有这些载体的细胞。
H.治疗用途
本发明提供的修饰的GH多肽和核酸分子可以用于治疗采用未修饰的GH治疗的任何状况。本节提供了修饰的GH多肽的举例用途和给药方法。这些描述的治疗是举例性的,并不限制GH的应用。
本发明提供了修饰的GH多肽,它们可用于采用GH治疗的各种治疗和诊断方法中。这些方法包括但不限于治疗下述生理性和医学状况的治疗。由于其改善的稳定性,本发明提供的修饰的GH多肽显示出在相应的体内活性和治疗效果中的改善。
特别地,修饰的GH多肽意在用于采用天然蛋白质进行治疗的治疗方法中。疾病的治疗可以包括但不限于生长缺陷疾病(包括但不限于特纳氏综合征、子宫内生长迟缓、特发性身材矮小症、PraderWilli综合征、地中海贫血)、AIDS消瘦、衰老、HIV感染对象的受损的免疫功能、分解代谢疾病(包括与呼吸衰竭和烧伤损伤相关的那些分解代谢疾病)、手术后恢复、充血性心肌病、肝移植、肝切除术后肝再生、慢性肾功能衰竭、肾性骨营养不良、骨质疏松、软骨发育不全/季肋发育不全、骨骼发育异常、慢性炎性或营养性疾病(如克罗恩氏病)、短肠综合征、青少年慢性关节炎、囊性纤维化、男性不育症、X连锁低磷血症性佝偻病、唐氏综合症、脊柱裂、努南综合征、肥胖、肌肉强度受损以及纤维肌痛。修饰的GH多肽和编码修饰的GH多肽的核酸分子也可以与其它治疗包括其它生物制剂和小分子化合物联合给药。
用修饰的GH多肽治疗疾病或病症可以通过使用本文所述的合适的配制品经任何合适的给药途径实现,包括但不限于皮下注射、口服、鼻、肺和透皮给药。如果需要,可以经验性确定或外推特定的剂量和时间以及治疗方案。例如,重组和天然GH多肽的示范剂量可以用作起始点以确定合适的剂量。更稳定的并具有增加的体内半衰期的修饰的GH多肽在降低的剂量或频率可以是有效的。本文提供用GH和重组形式进行治疗的剂量是示范剂量。但是这种示范剂量可以供选择GH变体的剂量方案参考。由于本发明提供的突变GH多肽展示增加的稳定性,所以剂量和给药方案可以与未修饰的生长激素的不同。具体的剂量和方案可以凭经验确定。
剂量水平对于本领域普通技术人员是清楚的并且可以基于各种因素如个体的体重、总的健康状况、年龄、采用的具体化合物的活性、性别、饮食、给药时间、排泄速率、药物组合、疾病或状况的严重性和过程、对象对疾病/状况的倾向以及治疗医生的判断等确定。可以与载体材料组合形成单剂型的活性成分的量根据被治疗的对象以及特定的给药模式而变化。
对象状况改善后,如果需要可以给予维持剂量的本文提供的化合物或组合物,并且剂量、剂型、或给药频率或它们的组合可以变化。在一些情况下,在疾病症状复发时对象可需要长期的间歇治疗。
1.生长缺陷
婴儿生长成成人是一个复杂过程,其中涉及各种基因和激素以及营养、饮食、锻练和休息。生长激素对于生长和发育是关键的并且是控制个体身高的主要激素。生长激素缺陷是一种通常由脑垂体或脑的下丘脑中发生问题而导致的疾病。生长激素缺陷可以在脑垂体中GH存在量不足或者GH存在量足够但是释放其所需的激素(GHRH)缺乏时发生。生长迟缓是一种医学状况,其中由于生长激素(GH)系统的缺陷,儿童的正常生长放慢或停止。
在儿童中有两种类型的GH缺陷:先天性和获得性。
先天性生长激素缺陷由于胎儿在子宫生长时脑垂体或下丘脑的问题导致,而获得性生长激素缺陷是当垂体和下丘脑附近区域以某种方式被损害时发生。在一些情况中,生长激素缺陷致病原因不明(“特发性”)。这类情况包括Prader-Willi综合征(一种特征为GH缺陷和身材矮小的先天性病症)、特纳氏综合征(一种仅在女孩中表现的特征为身材矮小的遗传缺陷)、慢性肾功能不全(肾功能失常(kidney malfunction),其可通常导致儿童生长迟缓)以及地中海贫血(一种遗传状况,特征在于血红蛋白合成失衡,导致严重贫血和畸形(malformed)血红细胞,由此可导致GH分泌降低和身材矮小)。
生长缺陷不仅影响儿童,对成人也是个大问题。成人中的GH缺陷是一种特殊的临床症状,具有无数的生理后果,包括但不限于:身体组成变化,包括向心性肥胖;血脂;肌肉强度;骨组成;运动能力(exercise capacity)和能量;心血管风险;和心理健康(例如,社会隔离和抑郁)。另外,研究表明患有垂体功能减退症的对象的心血管疾病死亡风险增加,可能是由于他们的GH缺陷导致。成人中的GH缺陷可以产生自垂体或垂体周围肿瘤,或是用于治疗这些状况的手术/放射的直接结果。较不常见地,成人中的GH缺陷由儿童时代获得的缺陷导致。
用于治疗生长缺陷治疗的重组GH补和/或替代应由机体正常产生的GH。对成人和儿童的治疗可包括全身性给予GH。例如,GH可以单独给药或者与其它治疗剂联合给药,联合给药可以在其它治疗剂给药之前、同时或之后进行。给药模式包括但不限于GH注射。本文描述的修饰的GH多肽和编码修饰的GH的核酸可用于生长缺陷治疗中。本文的修饰的GH展示增加的蛋白质稳定性和改善的半衰期,由此改善了药物组合物的治疗功效。因此,修饰的GH可以用于输送更持久更稳定的生长缺陷治疗。用修饰的GH治疗改善的例子包括例如但不限于剂量降低、给药更少次和/或频率下降、副作用下降以及治疗效果增加。
本发明提供的修饰的GH的剂量和方案可以经验性确定。例如,因为在性质如血清稳定性中的改善,剂量可以低于未修饰的GH的相应量。未修饰的GH的剂量可以用作指导确定修饰的GH的剂量。修饰的GH与未修饰的GH相比的活性水平和半衰期等因素可用于这类确定。
治疗生长缺陷的GH疗法的目的是长期GH替代至与相同性别和相似年龄的健康人类似的生理水平。GH的剂量方案可取决于多种因素,包括但不限于对象的年龄、青春期状态、对象耐受性和副作用发生率以及GH来源(是重组的还是天然的)。例如,重组GH的效力约为垂体GH效力的三分之一。特定的剂量和剂量方案可以经验性确定。对于患有GHD的儿童,未修饰的垂体GH的示范剂量可以是0.1mg/kg/wk(0.3IU/kg/wk);重组未修饰的GH的示范剂量可以是0.18-0.3mg/kg/wk(MacGillivray et al.Pediatrics 102:527-530(1998))。给予患有生长激素缺陷的儿童的平均GH剂量是0.3mg/kg/周,分成日剂量,皮下注射给药。治疗儿童中的GH缺陷病症的其它示范剂量包括对患有慢性肾功能不全的儿童为0.35 mg/kg/周;对患有特纳氏综合征的儿童为0.375mg/kg/周;对患有先天性身材矮小的儿童为0.3mg/kg/周;以及对患有子宫内生长迟缓的儿童为0.7mg/kg/周(参见,例如,Vance et al.The New England Journal ofMedicine 341(6):1206-1216(1999))。
成人中GH的起始剂量典型地是0.01-0.03mg/kg/wk,皮下注射。对于年龄不超过35岁的对象的最大日剂量典型地是0.18mg/kg/wk,老龄对象为0.09mg/kg/wk。在各种对生长激素替代的治疗研究中,未修饰的生长激素的剂量范围为约0.04至约0.18mg/kg/wk(see,Vance et al.The New England Journal of Medicine 341(6):1206-1216(1999))。其它人推荐(参见,例如the Growth HormoneResearch Society recommendations)起始剂量为105-210mg/周,而无论体重如何(参见,例如Consensus guidelines for the diagnosis andtreatment of adults with growth hormone deficiency:Summary statementof the Growth Hormone Research Society Workshop on Adults GrowthHormone Deficiency.J.Clin.Endocrinol.Metab.83:379-381(1998))。
2.恶病质
AIDS消瘦和其它形式的恶病质是指导致机体消耗生命肌肉和器官组织(瘦体重)而非主要依赖于机体脂肪供应来获得能量的代谢病症。患有AIDS消瘦的人典型地经历丧失5-10%或更多的瘦体重,这包括肌肉组织、身体器官、血细胞和淋巴液。尽管抗逆转录病毒治疗延长了HIV携带者的生命,但是AIDS消瘦仍是HIV/AIDS携带者不健康的一个主要原因。估计AIDS消瘦在4-30%的HIV感染个体中发生。研究证实当不治疗AIDS消瘦时,其直接与死亡率相关。在其它疾病例如癌症中观察到类似的消瘦表现。
本发明的修饰的GH的剂量和剂量方案可以经验性确定。未修饰的GH的剂量可以用作确定修饰的GH的剂量的指导。修饰的GH与未修饰的GH相比的活性水平和半衰期等因素可用于这类确定。本发明提供的修饰的GH具有增加的蛋白质稳定性和改善的半衰期,因此输送了治疗AIDS消瘦更持久更稳定的治疗效果。重组GH在1996年被美国FDA批准用于治疗AIDS消瘦。用于治疗恶病质的未修饰的生长激素治疗的推荐最初成人剂量不超过0.04mg/kg/周,分为6或7次皮下注射。根据个体对象的需要,剂量可以以4至8周间隔增加直至最大0.08mg/kg/周,这取决于对象对治疗的耐受性(参见Serostim_包装插页)。这些示范剂量可用作指导与修饰的GH与未修饰形式相比的性质和活性的确定一道确定修饰的GH多肽的剂量方案。
3.抗衰老
衰老与促性腺激素、促甲状腺素和垂体功能下降相关;通常被称为“生长停滞(somatopause)”。因此,作为一种垂体激素,GH也已经被报道在三十岁开始下降(参见例如Corpas et al. Endocr Rev 14:20-39(1993))。病理学降低的GH与伴随衰老可见的许多变化相关,包括脂肪增加、肌肉质量下降和骨质量下降。在生长激素缺陷的老龄个体中的生长激素替代能改善生活质量、增强骨和肌肉质量并且降低心血管风险。
如本文所述,本发明的修饰的GH的剂量和剂量方案可以经验性确定。本发明提供的修饰的GH的最初治疗剂量可以用未修饰的生长激素的批准的推荐最初成人剂量指导确定,之后根据修饰的GH的改善的治疗效果而调整确定。在61至81岁的老龄男性对象中重组hGH治疗的举例性皮下剂量可以是0.03mg/kg体重,每周注射三次,在早晨注射;注射之间间隔为一或两天(2.6IU/mg激素)(参见例如,Rudman et al.The New England Journal of Medicine 323(1):1-6(1990))。在老龄女性对象中重组hGH治疗的举例性皮下剂量可以是0.025mg/kg体重/天(Bonello et al.J.Am.Geriatr.Soc.44(9):1038-42(1996))。这些剂量可以伴随比较修饰的和未修饰的GH多肽的性质而确定修饰的GH的剂量来使用。例如,本发明提供的修饰的GH具有增加的蛋白质稳定性和改善的半衰期,由此输送更持久、更稳定的抗衰老治疗效果。
4.肾性骨营养不良
肾性骨营养不良包括各种骨骼病症,从高周转至低周转损害,两者均导致骨矿物质密度降低和骨折发生率增高,这是在患有慢性肾功能衰竭的对象中常见的。临床试验显示重组人GH治疗作为改善生长激素缺陷患者以及血透析的慢性肾病患者中的骨周转和骨矿物质密度的治疗的阳性效果(参见例如,Kotzmann et al.Journal ofNephrology 17(1):87-94(2004))。GH以及IGF-1对骨代谢和骨矿物质密度有显著效果。GH可刺激软骨细胞生长和功能并且通过刺激成骨细胞和破骨细胞和诱导胶原合成而直接或间接增加骨周转,由此增强长骨生长(long bone growth)。
本发明提供的修饰的GH的剂量和剂量方案可以经验性确定。本发明提供的修饰的GH的最初治疗剂量可以用未修饰的生长激素的批准的推荐最初成人剂量指导确定,之后根据修饰的GH的改善的治疗效果而调整确定。进行血透析的患有慢性肾功能衰竭的成人患者的举例性给药可以是在治疗的头4周在每次透析后每周皮下注射0.125IU/kg(40.5μg/kg)GH 3次,之后为0.25IU/kg(81μg/kg)。GH治疗的长度和剂量可以根据对象耐受性而变化(参见Kotzmann et al.Journal of Nephrology 17(1):87-94(2004))。修饰的GH与未修饰的GH的性质和活性的比较可用于确定交替的剂量和剂量方案。
5.囊性纤维化
患有囊性纤维化的对象、特别是儿童具有线性生长不良、增重不足和蛋白质分解代谢等问题。多项研究已经证实用GH治疗的儿童中身高和体重得以改善。其它研究显示GH治疗导致改善的用力肺活量、改善的运动耐量和骨积累。还有一些研究发现GH治疗改善了临床状态,这由住院治疗和静脉抗生素疗程降低而测得(参见例如,Hardin D.S.,Eur.J.Endocrinol.151(Suppl 1):S81-85(2004))。
本发明提供的修饰的GH的剂量和剂量方案可以经验性确定。例如,来自未修饰的GH的剂量和给药的指导以及修饰的GH与未修饰的GH的性质和活性的比较可以用于这种确定。患有囊性纤维化的儿科患者的举例性给药包括但不限于每日皮下GH注射,至0.3mg/kg/wk(see Hardin D.S.,Eur.J.Endocrinol.151(Suppl 1):S81-85(2004))。本发明提供的修饰的GH的最初治疗剂量可以是用未修饰的生长激素治疗的推荐最初成人剂量,之后根据本发明提供的修饰的GH的更长保持时间以及改善的治疗效果而调整确定。本发明提供的修饰的GH具有增加的蛋白质稳定性和改善的半衰期。这种修饰的GH可以在治疗患有囊性纤维化的对象中产生更持久的更稳定的治疗效果,并可以实现更低的剂量和降低的剂量频率。
6.其它状况
许多其它生理学或病理学状态也是GH治疗的潜在靶。这些生理学或病理学状态包括但不限于、应激、能量降低、体能下降、分解代谢疾病包括例如与呼吸衰竭和烧伤损伤相关的那些分解代谢疾病(参见例如,Hart et al.Ann.Surg.233(6):827-34(2001))、手术后恢复(参见例如,Yeo et al.Growth Horm.IGF Res.13(6):361-70(2003))、充血性心肌病(参见例如,Adamapolous et al.Eur.Heart J.24(24):2186-96(2003))、肝移植或肝切除术后的肝再生(参见例如Luo et al.World J Gastroenterol.10(9):1292-6(2004)),慢性炎性或营养性疾病如短肠综合征、克罗恩氏病(参见例如,Slonim et al.N.Engl.J.Med.342(22):1633-7(2000)),少年慢性关节炎(参见例如,Saha etal.J Rheumatol.1(7):1413-7(2004))、男性不育症(参见例如Ovesen etal.Fertil Steril.66(2):292-8(1996))和其它疾病如HIV感染对象的受损的免疫功能、骨质疏松、软骨发育不全/季肋发育不全、骨骼发育异常、X连锁低磷血症性佝偻病(参见例如,Seikaly et al.Pediatrics.100(5):879-84(1997))、努南综合征(参见例如Noordam et al.ActaPaediatr.90(8):889-94(2001))、肥胖症、唐氏综合症、脊柱裂、以及纤维肌痛(参见例如,Bennet et al.Am.J.Med.104(3):227-31(1998))。
I.制品和药盒
修饰的生长激素多肽和核酸可以被包装为制品,其含有包装材料、本文提供的有效用于治疗生长激素疾病或病症的修饰的GH多肽或编码修饰的GH或其衍生物或生物学活性部分的核酸分子、和指示修饰的GH或核酸分子用于治疗生长激素疾病或病症的标签。
本文提供的制品包含包装材料。用于包装药品的包装材料是本领域技术人员熟知的。参见例如U.S.Patent No.5,323,907,5,052,558和5,033,352。药物包装材料的例子包括但不限于泡罩包装(blisterpacks)、瓶、管、吸入器、泵、袋、小瓶、容器、注射器、瓶和适于选定的配方和给药和治疗模式的任何包装材料。本发明提供了许多种化合物和组合物的配方以及用于任何GH疾病或病症的各种治疗。
修饰的GH多肽和核酸分子也可以作为药盒(kit)提供。药盒可以包括修饰的GH和用于给药的项目。例如,修饰的GH可以用给药装置例如注射器、吸入器和施用器供应。药盒可以包括应用说明,包括剂量、剂量方案和给药模式说明。药盒还可包括修饰的GH和用于诊断的项目。例如,这类药盒包括测量浓度、生长激素的量或活性或对象的生长激素调节的系统的项目。
下述实施例仅为描述目的而包括进来,它们不限制本发明范围。
J.实施例
实施例1
在哺乳动物细胞表达质粒pNAUT中克隆编码GH的cDNA
在产生选择的突变之前,将编码GH蛋白的核酸分子(参见SEQID NO:1)克隆进哺乳动物表达载体。然后产生预先设计的定向突变体集合,每个突变体单独产生和加工,彼此物理分离地处于可寻址阵列中。
hGH-cDNA是通过用SMART试剂盒(Clontech)从人脑垂体mRNA(Clontech)体外合成而获得的。首先,用引物forward GH1(SEQ ID NO:702)和reverse GH1 REV(SEQ ID NO:703)经PCR扩增克隆编码hGH的cDNA。将PCR扩增产物克隆进大肠杆菌载体(pTOPO-TA)以产生命名为pTOPO-hGH4的质粒(SEQ ID NO:717)。
测序证实了编码hGH的cDNA的序列。用在克隆的每一端产生HindIII和XbaI限制位点的引物HGHFORHIND(SEQ ID NO:704)和HGHREV(SEQ ID NO:705)扩增测序的cDNA。用HindIII和XbaI消化后,将含有编码hGH的cDNA的PCR片段亚克隆进pUC-CMVhGHpA中的相应位点,产生pNAUT-hGH(SEQ ID NO:716)。
为表达hGH,用Herculase_(Invitrogen)DNA聚合酶,使用引物hGHFORPET(SEQ ID NO:706)和hGHREVPET(SEQ ID NO:707)经PCR扩增编码hGH的cDNA片段。引物分别产生正向和反向限制位点NdeI和BamHI。正向引物设计用于将脯氨酸2和脯氨酸5密码子从人密码子变为大肠杆菌密码子(CCA变为CCG以及CCC变为CCG),反向引物设计用于改变终止密码子(TAG变为TAA)以优化大肠杆菌产率。将PCR片段亚克隆进pET-24(Invitrogen;SEQ ID NO:718)中。为了优化大肠杆菌中的hGH产率,在hGH序列中进行额外的变化。使用诱变循环以将Arg 8和Arg 16密码子从人密码子变为细菌密码子,分别使用如下引物:hGHARG8FOR(SEQ ID NO:708)和hGHARG8REV(SEQ ID NO:709)以及hGHARG16FOR(SEQ IDNO:710)和hGHARG16REV(SEQ ID NO:711)。测序验证得到的构建体。最终质粒命名为pET-24Naut-hGH(参见SEQ ID No.718)。
实施例2
通过2D-扫描设计GH变体
使用本文描述的并且也在公开的申请US-2004-0132977-A1和U.S.申请系列号10/658,355中描述的2D-扫描被用于设计和获得具有改善的对蛋白酶解的抗性和/或改善的热耐受性的hGH突变体。is-HIT是基于如下鉴别:(1)待进化的蛋白质性质(即,对蛋白酶解的抗性或热耐受性);(2)氨基酸序列;和(3)各氨基酸的性质。
a.产生的用于hGH更高的对蛋白酶解的抗性的LEAD
使用2D-扫描设计变体。垂体hGH(SEQ ID NO:1)上的所选择的位置(is-HIT)是(编号相应于成熟蛋白质中的氨基酸位置):F1、P2、P5、L6、R8、L9、F10、D11、L15、R16、R19、L20、L23、F25、D26、Y28、E30、F31、E32、E33、Y35、P37、K38、E39、K41、Y42、F44、L45、P48、L52、F54、E56、P59、P61、R64、E65、E66、K70、L73、E74、L75、L76、R77、L80、L81、L82、W86、L87、E88、P89、F92、L93、R94、F97、L101、Y103、D107、Y111、D112、L113、L114、K115、D116、L117、E118、E119、L124、M125、R127、L128、E129、D130、P133、R134、F139、K140、Y143、K145、F146、D147、D153、D154、L156、K158、Y160、L162、L163、Y164、F166、R167、K168、D169、M170、D171、K172、E174、F176、L177、R178、R183、E186和F191。在每一个上述所列的并示于图1的is-HIT位置上的天然氨基酸用取代矩阵PAM250(图2)中定义的残基置换。所进行的实际残基取代见表3.总共产生了222个hGH变体。见表4和SEQ ID NO:2-223。
表3
is-HIT上的氨基酸 | 置换氨基酸 |
R | H,Q |
E | H,Q,N |
K | Q,N |
D | N,Q |
M | I,V |
P | A,S |
Y | I,H |
F | I,V |
W | H,S |
L | I,V |
is-HIT上的氨基酸(表3左栏)用选择的置换氨基酸(表3右栏)置换以产生具有增加的对蛋白酶解的抗性的GH变体。
表4
具有增加的对蛋白酶解的抗性hGH变体列表
F1I/P2A | F1I/P2S | F1V/P2A | F1V/P2S | P5A | P5S | L6I | L6V |
R8H | R8Q | L9I | L9V | F10I | F10V | D11N | D11Q |
M14I | M14V | L15I | L15V | R16H | R16Q | R19H | R19Q |
L20I | L20V | L23I | L23V | F25I | F25V | D26N | D26Q |
Y28H | Y28I | E30Q | E30H | E30N | F31I | F31V | E32Q |
E32H | E32N | E33Q | E33H | E33N | Y35H | Y35I | P37A |
P37S | K38N | K38Q | E39Q | E39H | E39N | K41N | K41Q |
Y42H | Y42I | F44I | F44V | L45I | L45V | P48A | P48S |
L52I | L52V | F54I | F54V | E56Q | E56H | E56N | P59A |
P59S | P61A | P61S | R64H | R64Q | E65Q | E65H | E65N |
E66Q | E66H | E66N | K70N | K70Q | L73I | L73V | E74Q |
E74H | E74N | L75I | L75V | L76I | L76V | R77H | R77Q |
L80I | L80V | L81I | L81V | L82I | L82V | W86H | W86S |
L87I | L87V | E88Q | E88H | E88N | P89A | P89S | F92I |
F92V | L93I | L93V | R94H | R94Q | F97I | F97V | L101I |
L101V | Y103H | Y103I | D107N | D107Q | Y111H | Y111I | D112N |
D112Q | L113I | L113V | L114I | L114V | K115N | K115Q | D116N |
D116Q | L117I | L117V | E118Q | E118H | E118N | E119Q | E119H |
E119N | L124I | L124V | M125I | M125V | R127H | R127Q | L128I |
L128V | E129Q | E129H | E129N | D130N | D130Q | P133A | P133S |
R134H | R134Q | F139I | F139V | K140N | K140Q | Y143H | Y143I |
K145N | K145Q | F146I | F146V | D147N | D147Q | D153N | D153Q |
D154N | D154Q | L156I | L156V | L157I | L157V | K158N | K158Q |
Y160H | Y160I | L162I | L162V | L163I | L163V | Y164H | Y164I |
F166I | F166V | R167H | R167Q | K168N | K168Q | D169N | D169Q |
M170I | M170V | D171N | D171Q | K172N | K172Q | E174Q | E174H |
E174N | F176I | F176V | L177I | L177V | R178H | R178Q | R183H |
R183Q | E186Q | E186H | E186N | F191I | F191V |
b.产生的用于hGH的更高的热耐受性的LEAD
表5示出了使用2D-扫描在hGH上鉴别的用于增加的热耐受性的is-HIT列表。鉴别了16个位置(is-HIT),它们的编号相应于成熟蛋白质中的编号。选择了is-HIT靶位置后,鉴别用于每个is-HIT靶位置的置换氨基酸。产生了特异于每个is-HIT的合适的置换氨基酸,它们保持或改善所需的蛋白质生物学活性(即GH活性)并且增加蛋白质稳定性。
通过一个接一个置换在特异的is-HIT靶位置的单氨基酸残基而进行诱变。产生的每个突变体是一个诱变反应的单一产物。取代的氨基酸与蛋白质结构和功能相容。为了选择针对每一is-HIT位置的候选置换氨基酸,使用了氨基酸取代矩阵。表5所示的每一is-HIT位置的天然氨基酸用增加与其它氨基酸的极性相互作用的残基E、D、K、R、N、Q、S和T置换。产生了总共128个hGH变体(见表6和SEQ ID NO:224-351)。
在hGH(SEQ ID NO:1)的螺旋A和C中鉴别的用以增加热耐受性的is-HIT位置是L6、L9、A13、L15、A17、L20、L23、A24、A105、V110、L113、L114、L117、I121、L124、L128(见表5)。选择特异的氨基酸(包括例如谷氨酸(E)、天冬氨酸(D)、赖氨酸(K)和精氨酸(R))作为置换每一is-HIT位置的氨基酸的氨基酸,由此在螺旋A和C的区域中导入额外的螺旋间键(见表6)。
表5
hGH增加的热耐受性的is-HIT列表
L6 | L9 |
A13 | L15 |
A17 | L20 |
L23 | A24 |
A105 | V110 |
L113 | L114 |
L117 | I121 |
L124 | L128 |
表6
增加的热耐受性的hGH突变体列表
L6E | L6D | L6K | L6R | L6N | L6Q | L6S | L6T |
L9E | L9D | L9K | L9R | L9N | L9Q | L9S | L9T |
A13E | A13D | A13K | A13R | A13N | A13Q | A13S | A13T |
L15E | L15D | L15K | L15R | L15N | L15Q | L15S | L15T |
A17E | A17D | A17K | A17R | A17N | A17Q | A17S | A17T |
L20E | L20D | L20K | L20R | L20N | L20Q | L20S | L20T |
L23E | L23D | L23K | L23R | L23N | L23Q | L23S | L23T |
A24E | A24D | A24K | A24R | A24N | A24Q | A24S | A24T |
A105E | A105D | A105K | A105R | A105N | A105Q | A105S | A105T |
V110E | V110D | V110K | V110R | V110N | V110Q | V110S | V110T |
L113E | L113D | L113K | L113R | L113N | L113Q | L113S | L113T |
L114E | L114D | L114K | L114R | L114N | L114Q | L114S | L114T |
L117E | L117D | L117K | L117R | L117N | L117Q | L117S | L117T |
I121E | I121D | I121K | I121R | I121N | I121Q | I121S | I121T |
L124E | L124D | L124K | L124R | L124N | L124Q | L124S | L124T |
L128E | L128D | L128K | L128R | L128N | L128Q | L128S | L128T |
实施例3
诱变
设计了一系列诱变引物以如下所述在hGH cDNA中产生合适的位点特异性突变。诱变反应使用Quickchange_试剂盒(Invitrogen)用pNaut-hGH(SEQ ID NO:716)作为模板进行。每个诱变反应含有一对诱变引物(有义和反义)(SEQ ID NO:352-701)。对于每个反应,将10皮摩尔的每个有义诱变引物与30ng模板、10皮摩尔反义诱变引物和1.25U Pfu turbo聚合酶(Invitrogen)混和。为使DNA退火,将PCR平板在95℃保温30秒。在14个循环的95℃30秒、50℃1分钟和68℃8分钟中进行延伸和连接反应。为了消除亲代质粒,用5U DpnI(BioLabs)进行限制消化。感受态细胞的转化用20μl感受态细胞(HT-96,Novagen)和1ng DNA完成。质粒DNA分离用Nucleobond_96turbo Minipreps试剂盒(Macherey-Nagel)进行。突变质粒的选择用100μg/ml氨苄青霉素进行。真核表达质粒的转化体用氨苄青霉素选择,原核表达质粒的转化体用卡那霉素进行选择。测序证实被导入而产生这一编码蛋白质的候选LEAD hGH突变质粒集合的每一突变。hGH的LEAD突变体首先在pNaut-hGH质粒中产生。使用相应的引物(SEQ ID NO:352-701)在原核表达质粒(pET-hGH)中再次产生突变体。突变的存在用测序验证。
表7
产生的突变体 | |||||||
F1I/P2A | F25I | Y42H | K70Q | F92V | E118H | K145N | K168N |
F1I/P2S | F25V | Y42I | L73I | L93I | E118N | K145Q | K168Q |
FIV/P2A | D26N | F44I | L73V | L93V | E119Q | F146I | D169N |
F1V/P2S | D26Q | F44V | E74Q | R94H | E119H | F146V | D169Q |
P5A | Y28H | L45I | E74H | R94Q | E119N | D147N | M170I |
P5S | 728I | L45V | E74N | F97I | L124I | D147Q | M170V |
L6I | E30Q | P48A | L75I | F97V | L124V | D153N | D171N |
L6V | E30H | P48S | L75V | L101I | M125I | D153Q | D171Q |
R8H | E30N | L52I | L76I | L101V | M125V | D154N | K172N |
R8Q | F31I | L52V | L76V | Y103H | R127H | D154Q | K172Q |
L9I | F31V | F54I | R77H | Y103I | R127Q | L156I | E174Q |
L9V | E32Q | F54V | R77Q | D107N | L128I | L156V | E174H |
F10I | E32H | E56Q | L80I | D107Q | L128V | L157I | E174N |
F10V | E32N | E56H | L80V | Y111H | E129Q | L157V | F176I |
D11N | E33Q | E56N | L81I | Y111I | E129H | K158N | F176V |
D11Q | E33H | P59A | L81V | D112N | E129N | K158Q | L177I |
M14I | E33N | P59S | L82I | D112Q | D130N | Y160H | L177V |
M14V | Y35H | P61A | L82V | L113I | D130Q | Y160I | R178H |
L15I | Y35I | P61S | W86H | L113V | P133A | L162I | R178Q |
L15V | P37A | R64H | W86S | L114I | P133S | L162V | R183H |
R16H | P37S | R64Q | L87I | L114V | R134H | L163I | R183Q |
R16Q | K38N | E65Q | L87V | K115N | R134Q | L163V | E186Q |
R19H | K38Q | E65H | E88Q | K115Q | F139I | Y164H | E186H |
R19Q | E39Q | E65N | E88H | D116N | F139V | Y164I | E186N |
L20I | E39H | E66Q | E88N | D116Q | K140N | F166I | F191I |
L20V | E39N | E66H | P89A | L117I | K140Q | F166V | F191V |
L23I | K41N | E66N | P89S | L117V | Y143H | R167H | |
L23V | K41Q | K70N | F92I | E118Q | Y143I | R167Q |
实施例4
在哺乳动物细胞中产生天然和修饰的hGH多肽
将HEK 293 EBNA细胞(例如可得自Roche;也参见Kruyt et al.Blood 90:3288-3295(1997))在补加10%SVF和遗传霉素的Dulbecco’s MEM-GlutamaxI-丙酮酸钠培养基中培养。细胞在37℃于7%CO2气氛下生长。通过瞬时转染产生天然或突变hGH。细胞以5×105细胞/孔接种在6孔板中的补加1%SVF的DMEM中;40小时后,每种蛋白质用PEI(25 KDa;Sigma-Aldrich)转染2个孔中的细胞;2个孔模拟转染以产生用于ELISA和用于确定生物学活性的阴性对照。24小时后,收获上清,样品等分在96孔板中并储存在-20℃用于经ELISA标准化(hGH ELISA,Roche)或用于筛选。
实施例5
分析修饰的hGH突变体多肽的生物学活性
为进行初步筛选,以平行的至少两个标准逐个测试每个突变体:(i)hGH生物学活性(基于细胞的体外测定,参见下述);(ii)对蛋白酶解的抗性;(iii)热耐受性。对于所有测试,产生系列稀释曲线,并获得针对每一个突变体的“EC50”值。产生天然hGH(SEQ ID NO:1)并用与hGH突变体相同的方案处理,并在整个过程中用作参考标准。得自英国的NIBSC的国际hGH标准也用作另一个参考标准。所有处理和测试均一式三份进行。
a.细胞增殖测定
测量每一突变体的体外标准GH生物学活性并与天然hGH(SEQ ID NO:1)比较在Nb2-11C细胞(大鼠淋巴瘤细胞系)上的细胞增殖活性。测定细胞增殖活性的剂量(浓度)-应答(活性)实验使得能计算每一突变体的生物学活性“效力(potency)”或EC50。在与蛋白酶解样品如特异的蛋白酶、选定的蛋白酶的混合物、人血清或人血液保温后测量同一系统中的细胞增殖活性。与蛋白酶解样品保温后活性的评价使得可以确定在暴露于蛋白酶后的残留细胞增殖活性和相应的半衰期动力学。
用Nb2-11细胞(大鼠淋巴母细胞)经增殖细胞测定测量暴露于蛋白酶后的剩余生物学活性。将Nb2-11细胞培养在补加10%SVF和10%马血清(ES)的Fisher培养基中。增殖测定之前24小时,离心细胞并用PBS洗涤。细胞然后在仅补加10%ES的Fisher培养基中以0.5-0.8×106细胞/ml的密度培养。培养24小时之后,将细胞以4×104细胞/孔接种在96孔板中并用3倍系列稀释(12个点)的在6000pg/ml和0.3 pg/ml之间的处理前的天然hGH或突变体处理,一式三份进行。NIBSC hGH用作每一增殖测定的内部对照。
用天然或突变hGH(用蛋白酶处理前的)处理48小时后,测量Nb2-11细胞的增殖。每孔加入20μl Cell Titer 96 AQ(Promega)并在37℃保温细胞1小时。所述测定测量四氮唑MTS向可溶的甲_的转变。样品在Spectramax reader(Molecular Device)中于490nm读数。
b.蛋白酶解抗性测定
在经ELISA确定了产生的hGH(天然和每个突变hGH)的量之后,含有15ng hGH或变体的高至150μl的上清用1%w/w上清中的总蛋白的量的蛋白酶混合物处理(1%血清,因为1%血清中的总蛋白浓度是600μg/ml,所有1%蛋白酶是基于蛋白质的总量而不仅是hGH的量)。用蛋白酶处理hGH突变体以鉴别抗性分子。突变hGH分子与天然hGH相比的相对抗性经暴露于(120min,25℃)蛋白酶混合物(含有1.5pg每种下述蛋白酶(1%wt/wt,Sigma):α-胰凝乳蛋白酶、羧肽酶、内切蛋白酶Arg-C、内切蛋白酶Asp-N、内切蛋白酶Glu-C、内切蛋白酶Lys-C和胰蛋白酶)而确定。在保温时间结束时,向每一反应中加入10μl抗蛋白酶完整培养基以抑制蛋白酶活性,所述培养基含有在10ml DMEM中1/1000稀释的mini EDTA free tablets(Roche)。处理的样品随后用于确定残留活性。
随暴露于蛋白酶的时间的残留hGH活性百分比用1.5pg蛋白酶混合物经动力学研究评价。对于每一新的测定,蛋白酶混合物从内切蛋白酶Glu-C(SIGMA)200μg/ml、胰蛋白酶(SIGMA)400μg/ml和α-胰凝乳蛋白酶(SIGMA)400μg/ml的原液新鲜制备。保温时间是(小时):0、0.08、0.25、0.5、0.75、1、2、4、6和8。简而言之,将20μl每种蛋白酶解样品(蛋白酶、血清、血液)加入到100μl 400pg/ml和800pg/ml的hGH中,保温如所指出的不同时间。在合适的时间点,向每孔中加入10μl抗蛋白酶混合物、完整mini EDTA-freeprotease inhibitor cocktail(Roche;1片溶解于10ml DMEM中然后稀释1/500)以终止蛋白酶解反应。然后对每一样品进行所述的生物学活性测定以确定每一时间点的残余活性。
c.热耐受性测定
为了评价热耐受性的动力学,增加温度保温增加时间点而测试各个hGH变体。经ELISA确定所产生的蛋白质(针对各hGH变体和天然hGH)的量之后,将0.4ng天然hGH或修饰的hGH加入到250μl补加1X抗蛋白酶鸡尾酒混合物(mini EDTA free,Roche)的DMEM无血清培养基中并在深孔板中于37℃保温。
在增加的时间点(0,2,4,6,8,12,24,36,48小时),将380μl补加5%SVF的DMEM培养基加入到20μl等份中(hGH终浓度为12000pg/ml)。立即冷冻样品并储存在-20℃。
对于每个分子(天然hGH或突变hGH),确定在37℃的每一保温时间点的残余生物学活性(在NB2-11C细胞上的细胞增殖测定;见上述)。
实施例6
GH变体的评价
用一种数学模型和算法(NautScan;法国专利9915884;还参见公开的国际PCT申请WO 01/44809,其基于PCT No.PCT/FR00/03503,见上述)分析各种生物学活性,包括蛋白酶抗性和每个突变体的效力(potency)。
用基于Hill方程的模型处理数据,该模型使用每个突变体的性能的关键特征指示符。简而言之,Hill方程是一种将药物(即测试化合物或物质)浓度与测量的应答联系起来的数学模型。
其中y是测量的变量,如应答、信号,ymax是可实现的最大应答,[D]是药物(例如GH或修饰的GH)的摩尔浓度,[D50]是产生对药物的50%最大应答的浓度,n是斜率参数,如果药物结合单位点并且位点之间或之中无协同性则n是1。Hill图是配体占据的受体与游离受体之比的log10 vs.log[D](M)。斜率是n,其中斜率大于1表示结合位点中的协同性,斜率小于1可以表示结合的异质性。这一方程已被用于评价复杂生物学系统中的相互作用的方法中,参数π,κ,τ,ε,η,θ如下:
π 是作用于测定(基于细胞的)系统上的生物制剂的效力;
κ 是测定系统引起对生物制剂的应答的抗性常数;
ε 是在Hill曲线(或者通常任何其它S形或线性逼近)的拐点的斜率,用于估算引起生物学或药理学应答的整体反应(生物制剂和测定系统一起)的效率
τ 用于测量生物制剂的有限稀释或表观效价;
θ 用于测量生物制剂的绝对有限稀释或效价;以及
η 是生物学过程或反应的异质性。η测量的是整体反应中存在的非连续相,其由Hill系数值或抗性常数中的陡变反映。
修饰的GH多肽基于它们的由EC50评价的性能的值而排列。用突变体的系列稀释度在基于细胞的增殖测定(见实施例5)中确定每一突变体的生物学比活性(即每单位蛋白质质量的生物学活性;单位/mg蛋白质)。对每一曲线制备12个稀释度并一式三份测定每一稀释度。使用来自系列稀释测定的数据,获得了达到50%活性所需的浓度(EC50)。如下所述使用集成在NEMO(Newly evolvedMolecules)软件(Nautilus Biotech)中的Gnuplot 5.0(绘制数据曲线的软件;在线得自gnuplot.info)将实验点拟合成S形曲线。用于S形曲线拟合的方程是:
Sig(x)=base+pmax*(x exp.nu)/X exp.nu)+kappa
其中base,pmax,nu和kappa是针对每一曲线的参数,如下:base=0.1,pmax=1,kappa=5000,nu=2.5,n为5-150之间。Gnuplot重复拟合“n”次直至其发现适合实验数据最佳同时每个实验点和拟合曲线上的理论点之间的距离的平方之和最小的曲线。一旦获得针对每一突变体的拟合曲线,则从曲线计算相应的EC50和比活性。举例性的曲线如图3所示。
处于列表中的前列的那些被选择作为LEAD。表8提供了所述的所有候选LEAD突变体的与天然hGH(SEQ ID NO:1以及本文所述的NIBSC GH标准)相比的生物学活性(EC50)和对蛋白酶解的抗性(标为“无变化”、“增加”或“降低”)数据。EC50计算是基于两次测定的平均值。修饰的hGH蛋白质的EC50与野生型hGH多肽相比整体上是降低的。
表8
3个测定的平均值 | 3个测定的平均值 | ||||
突变 | EC50 | 对蛋白酶的抗性 | 突变 | EC50 | 对蛋白酶解的抗性 |
F1I/P2A | 90.56 | 增加 | F92V | 97.72 | 无变化 |
F1I/P2S | 120.67 | 无变化 | L93I | 76.97 | 无变化 |
F1V/P2A | 75.28 | 无变化 | L93V | 96.40 | 无变化 |
F1V/P2S | 60.39 | 无变化 | R94H | 88.74 | 无变化 |
P5A | 83.66 | 无变化 | R94Q | 94.03 | 无变化 |
P5S | 99.51 | 增加 | F97I | 93.38 | 无变化 |
L6I | 92.29 | 无变化 | F97V | 68.32 | 无变化 |
L6V | 111.32 | 无变化 | L101I | 94.59 | 无变化 |
R8H | 85.11 | 无变化 | L101V | 134.99 | 增加 |
R8Q | 108.30 | 无变化 | Y103H | 83.33 | 无变化 |
L9I | 93.41 | 无变化 | Y103I | 88.96 | 无变化 |
L9V | 119.78 | 增加 | D107N | 87.43 | 无变化 |
F10I | 119.48 | 无变化 | D107Q | 70.59 | 无变化 |
F10V | 100.49 | 无变化 | Y111H | 58.16 | 无变化 |
D11N | 100.47 | 增加 | Y111I | 56.19 | 增加 |
D11Q | 72.23 | 无变化 | D112N | 73.94 | 增加 |
M14I | 103.28 | 无变化 | D112Q | 72.75 | 无变化 |
M14V | 81.47 | 增加 | L113I | 83.87 | 无变化 |
L15I | 76.31 | 无变化 | L113V | 82.49 | 无变化 |
L15V | 2,701.08 | 无变化 | L114I | 60.48 | 无变化 |
R16H | 96.75 | 增加 | L114V | 81.81 | 无变化 |
R16Q | 104.24 | 无变化 | K115N | 97.38 | 无变化 |
R19H | 59.89 | 无变化 | K115Q | 89.44 | 无变化 |
R19Q | 129.68 | 无变化 | D1 16N | 75.90 | 无变化 |
L20I | 91.80 | 无变化 | D116Q | 99.39 | 增加 |
L20V | 105.58 | 无变化 | L117I | 126.24 | 无变化 |
3个测定的平均值 | 3个测定的平均值 | ||||
突变 | EC50 | 对蛋白酶的抗性 | 突变 | EC50 | 对蛋白酶解的抗性 |
L23I | 95.68 | 增加 | L117V | 111.76 | 无变化 |
L23V | 95.33 | 增加 | E118Q | 93.67 | 无变化 |
F25I | 75.71 | 无变化 | E118H | 101.91 | 无变化 |
F25V | 61.20 | 无变化 | E118N | 83.61 | 无变化 |
D26N | 77.15 | 增加 | E119Q | 110.30 | 增加 |
D26Q | 84.56 | 无变化 | E119H | 79.50 | 无变化 |
Y28H | 125.40 | 无变化 | E119N | 73.04 | 无变化 |
Y28I | 100.63 | 无变化 | L124I | 99.38 | 无变化 |
E30Q | 67.36 | 无变化 | L124V | 103.17 | 增加 |
E30H | 64.49 | 无变化 | M125I | 97.04 | 增加 |
E30N | 61.53 | 无变化 | M125V | 112.09 | 增加 |
F31I | 146.71 | 无变化 | R127H | 88.81 | 无变化 |
F31V | 103.13 | 无变化 | R127Q | 72.34 | 无变化 |
E32Q | 71.39 | 无变化 | L128I | 137.41 | 无变化 |
E32H | 50.23 | 无变化 | L128V | 86.77 | 无变化 |
E32N | 70.15 | 无变化 | E129Q | 96.71 | 无变化 |
E33Q | 77.59 | 无变化 | E129H | 114.09 | 无变化 |
E33H | 83.78 | 无变化 | E129N | 77.15 | 无变化 |
E33N | 75.98 | 无变化 | D130N | 107.61 | 无变化 |
Y35H | 104.16 | 无变化 | D130Q | 176.58 | 无变化 |
Y35I | 93.11 | 无变化 | P133A | 102.73 | 增加 |
P37A | 113.48 | 无变化 | P133S | 123.01 | 无变化 |
P37S | 71.11 | 无变化 | R134H | 93.22 | 增加 |
K38N | 121.66 | 增加 | R134Q | 76.53 | 无变化 |
K38Q | 127.34 | 无变化 | F139I | 89.02 | 无变化 |
E39Q | 78.13 | 无变化 | F139V | 95.71 | 无变化 |
E39H | 73.37 | 无变化 | K140N | 92.18 | 增加 |
3个测定的平均值 | 3个测定的平均值 | ||||
突变 | EC50 | 对蛋白酶的抗性 | 突变 | EC50 | 对蛋白酶解的抗性 |
E39N | 94.38 | 无变化 | K140Q | 69.56 | 无变化 |
K41N | 177.74 | 无变化 | Y143H | 68.,24 | 无变化 |
K41Q | 237.04 | 增加 | Y143I | 71.15 | 无变化 |
Y42H | 146.32 | 增加 | K145N | 81.32 | 无变化 |
Y42I | 244.83 | 增加 | K145Q | 109.72 | 无变化 |
F44I | 140.34 | 无变化 | F146I | 90.18 | 无变化 |
F44V | 221.00 | 无变化 | F146V | 105.26 | 无变化 |
L45I | 110.10 | 无变化 | D147N | 143.77 | 增加 |
L45V | 164.03 | 无变化 | D147Q | 61.81 | 增加 |
P48A | 110.11 | 无变化 | D153N | 62.48 | 增加 |
P48S | 90.70 | 无变化 | D153Q | 101.35 | 无变化 |
L52I | 78.15 | 无变化 | D154N | 89.30 | 无变化 |
L52V | 78.87 | 无变化 | D154Q | 74.20 | 无变化 |
F54I | 93.14 | 无变化 | L156I | 116.04 | 增加 |
F54V | 87.81 | 无变化 | L156V | 116.93 | 无变化 |
E56Q | 87.23 | 增加 | L157I | 83.02 | 增加 |
E56H | 100.61 | 无变化 | L157V | 79.94 | 无变化 |
E56N | 70.11 | 增加 | K158N | 186.86 | 增加 |
P59A | 101.11 | 无变化 | K158Q | 100.86 | 无变化 |
P59S | 94.22 | 无变化 | Y160H | 111.76 | 无变化 |
P61A | 99.87 | 无变化 | Y160I | 111.35 | 无变化 |
P61S | 134.30 | 无变化 | L162I | 80.80 | 增加 |
R64H | 98.68 | 无变化 | L162V | 94.70 | 无变化 |
R64Q | 116.96 | 无变化 | L163I | 96.11 | 无变化 |
E65Q | 61.83 | 增加 | L163V | 75.72 | 无变化 |
E65H | 106.92 | 无变化 | Y164H | 94.71 | 无变化 |
E65N | 142.94 | 无变化 | Y164I | 95.73 | 无变化 |
3个测定的平均值 | 3个测定的平均值 | ||||
突变 | EC50 | 对蛋白酶的抗性 | 突变 | EC50 | 对蛋白酶解的抗性 |
E66Q | 81.38 | 增加 | F166I | 105.21 | 增加 |
E66H | 72.06 | 无变化 | F166V | 72.90 | 无变化 |
E66N | 142.65 | 无变化 | R167H | 189.52 | 增加 |
K70N | 107.42 | 无变化 | R167Q | 283.50 | 增加 |
K70Q | 94.00 | 无变化 | K168N | 139.82 | 增加 |
L73I | 80.51 | 无变化 | K168Q | 126.42 | 增加 |
L73V | 110.40 | 增加 | D169N | 48.74 | 无变化 |
E74Q | 112.51 | 无变化 | D169Q | 105.62 | 增加 |
E74H | 120.05 | 无变化 | M170I | 89.01 | 无变化 |
E74N | 143.00 | 增加 | M170V | 78.96 | 无变化 |
L75I | 114.32 | 无变化 | D171N | 80.78 | 增加 |
L75V | 114.22 | 无变化 | D171Q | 84.85 | 增加 |
L76I | 92.75 | 无变化 | K172N | 712.32 | 无变化 |
L76V | 128.45 | 无变化 | K172Q | 127.47 | 增加 |
R77H | 199.12 | 无变化 | E174Q | 62.40 | 增加 |
R77Q | 77.86 | 无变化 | E174H | 75.14 | 增加 |
L80I | 110.78 | 无变化 | E174N | 78.32 | 增加 |
L80V | 80.57 | 无变化 | F176I | 146.43 | 无变化 |
L81I | 175.02 | 无变化 | F176V | 110.75 | 无变化 |
L81V | 132.13 | 增加 | L177I | 126.12 | 增加 |
L82I | 111.14 | 无变化 | L177V | 102.88 | 增加 |
L82V | 80.58 | 无变化 | R178H | 77.96 | 无变化 |
W86H | 70.90 | 无变化 | R178Q | 119.45 | 增加 |
W86S | 100.31 | 无变化 | R183H | 134.43 | 无变化 |
L87I | 226.80 | 无变化 | R183Q | 82.50 | 无变化 |
L87V | 89.48 | 增加 | E186Q | 76.54 | 无变化 |
E88Q | 61.89 | 无变化 | E186H | 88.28 | 无变化 |
3个测定的平均值 | 3个测定的平均值 | ||||
突变 | EC50 | 对蛋白酶的抗性 | 突变 | EC50 | 对蛋白酶解的抗性 |
E88H | 401.37 | 无变化 | E186N | 110.73 | 无变化 |
E88N | 100.23 | 无变化 | F191I | 125.72 | 增加 |
P89A | 113.76 | 无变化 | F191V | 115.73 | 无变化 |
P89S | 89.65 | 无变化 | hGH WT | 125.69 | |
F92I | 101.81 | 无变化 | hGH NIBSC | 135.20 |
实施例7
人类生长激素在大鼠中的药动学研究、
a.hGH多肽的制备
a.1细胞培养
Escherichia coli菌株BLR是BL21的recA-衍生菌株,其改善质粒单体的产率并能帮助稳定含有重复序列的靶质粒(Novagen)。
a.2 Vector:Modified pET-24a
pET-24a(+)载体(Novagen)携带N-末端T7-Tag_序列、任选的C-末端His-Tag_序列和选择标记卡那霉素。hGH cDNA序列被导入位点NdeI/BamHI。His-Tag_和F1复制起点已从这一载体被除去。将E.coli菌株BLR(Novagen)用pET24Naut-hGH载体(SEQ ID NO:719)转化并在加有30μg/ml卡那霉素(Sigma)的4%LB培养液(LB-BrothPowder,Gibco)中于37℃培养。
a.3 过表达诱导
过夜培养后,将细菌稀释至OD600 0.07并培养至OD600 0.6(约8.7×106细胞/mL)。hGH cDNA序列的过表达通过加入0.5M IPTG(Amersham)3小时而诱导(估测的倍增时间为45分钟)。最终OD600在1.8至2.2之间(SOP-PRD-001/02),产量估测为约30-40mg/L。
a.4 取细胞、分离和破碎
由于在大肠杆菌中过表达以及该蛋白质的性质(大的疏水面积),发现hGH蛋白在包涵体中。通过用溶菌酶(BoehringerMannheim,Ref.837059)、10mM CaCl2(Calcium Chloride DihydrateSigma Ultra)和1U/ml DNase I(DNaseI,RNase free,10U/μl,Roche)裂解细菌而从细胞中释放包涵体。包涵体通过在10,000g离心15min而收获,本步骤收率(step yield)为90%,估测的纯度为>60%。纯度是通过组合考马斯蓝染色的SDS-PAGE和hGH ELISA试剂盒(hGH Elisa kit,Roche)而估测的。包涵体的脂质污染通过使用TritonX-100 0.5%(Carlo Erba)洗两次随后用甘油5%(Sigma Ref,G-5150)洗2次的标准程序而解决。本步骤收率是90%,纯度约为>70%。
a.5 产物溶解和再折叠
用6M盐酸胍以5∶1体积进行变性过程30分钟,随后通过用Tris 50mM pH 8,EDTA 10mM,L-Arg 0.2M(Sigma)和5%甘油稀释至50∶1体积当量而再折叠。通过首先在20x体积比的相同缓冲液中透析过夜随后在Tris pH 8 10mM缓冲液中以80X体积比第二次透析(重复两次)而去除过量的盐酸胍。本步骤收率为80%,估测的纯度为95%。
a.6 层析
用阴离子交换柱(IEC)Q-sepharose FF 20mL(Amersham)以流速120cm/hr在50mM Tris(pH 8)平衡/加样条件于4℃进行层析,随后在50mM Tris(pH 8)和200mM NaCl中洗脱。本步骤收率为80%,估测的纯度为>90%。用Affinity Pak Detoxy-Gel EndotoxinRemoving Gel(Pierce,Prod#20344)实现可容忍的内毒素水平。
b. 体内研究
在雄性Sprague-Dawley大鼠(6只大鼠/分子)中测试hGH突变体和天然hGH蛋白质的药动学谱。测试的LEAD是:F1I/P2A、D11N、M14V、R16H、L23I、L23V、K41Q、Y42H、Y42I、L81V、L101V、D116Q、E119Q、M125I、M125V、K140N、E174N、L177I、R178Q和F191I,与野生型相比。给药为皮下(SC)注射2mg hGH/kg大鼠体重。在15分钟(min)和96小时(hrs)之间增加的时间点取血样,并以斑点印迹集合在Whatman No.2滤纸上。在48小时期间于4℃从5mm2滤纸上制备在PBS中的hGH蛋白质洗脱液。洗脱的hGH量用特异于hGH的商购ELISA(Roche)确定。
野生型hGH多肽在注射后3小时被降解,而修饰的hGH在注射后24小时和30小时仍在血清中被发现。因此本发明提供的修饰的hGH多肽比野生型hGH多肽在体内是更蛋白酶解稳定的。
由于修饰对于本领域技术人员而言是明显的,因此本发明仅由所附权利要求书的范围而限制。
序列表
<110>诺逖勒斯生物技术公司
<120>修饰的生长激素
<130>17109-015WO1(925PC)
<140>Unassigned
<141>Herewith
<150>60/706,697
<151>2005-08-08
<150>60/625,652
<151>2004-11-04
<160>719
<170>FastSEQ for Windows Version 4.0
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<211>191
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<213>Homo Sapiens
<220>
<223>Wild type human Growth Hormone
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Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 11
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 11
Phe Pro Thr Ile Pro Leu Ser Gln Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 12
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 12
Phe Pro Thr Ile Pro Leu Ser Arg Ile Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 13
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 13
Phe Pro Thr Ile Pro Leu Ser Arg Val Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 14
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 14
Phe Pro Thr Ile Pro Leu Ser Arg Leu Ile Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 15
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 15
Phe Pro Thr Ile Pro Leu Ser Arg Leu Val Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 16
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 16
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asn Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 17
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 17
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Gln Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 18
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 18
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Ile Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 19
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 19
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Val Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
80 185 190
<210> 20
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 20
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Ile Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 21
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 21
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Val Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 22
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 22
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu His
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 23
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 23
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Gln
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 24
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 24
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His His Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 25
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 25
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Gln Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 26
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 26
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Ile His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 27
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 27
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Val His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 28
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 28
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Ile Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 29
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 29
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Val Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 30
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 30
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Ile Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 31
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 31
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Val Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 32
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 32
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asn Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 33
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 33
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Gln Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 34
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 34
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr His Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 35
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 35
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Ile Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 36
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 36
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Gln Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 37
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 37
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln His Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 38
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 38
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Asn Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 39
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 39
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Ile Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala ASn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 40
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 40
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Val Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 41
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 41
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Gln
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 42
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 42
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe His
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 43
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 43
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Asn
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 44
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 44
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Gln Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 45
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 45
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
His Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 46
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 46
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Asn Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 47
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 47
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala His Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 48
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 48
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Ile Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 49
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 49
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Ala Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 50
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 50
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Ser Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 51
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 51
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Asn Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 52
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 52
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Gln Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 53
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 53
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Gln Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 54
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 54
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys His Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 55
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 55
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Asn Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 56
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 56
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Asn Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 57
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 57
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Gln Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 58
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 58
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys His Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 59
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 59
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Ile Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 60
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 60
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Ile Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 61
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 61
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Val Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 62
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 62
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Ile Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 63
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 63
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Val Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 64
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 64
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Ala
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 65
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 65
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Ser
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 66
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 66
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Ile Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Ash Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 67
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 67
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Val Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 68
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 68
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Ile Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 69
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 69
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Val Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 70
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 70
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Gln Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 71
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 71
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser His Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 72
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 72
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Asn Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 73
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 73
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Ala Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 74
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 74
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Ser Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 75
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 75
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Ala Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 76
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 76
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Ser Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 77
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 77
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn His
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 78
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 78
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Gln
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 79
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 79
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Gln Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 80
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 80
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
His Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 81
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 81
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Asn Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 82
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 82
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Gln Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 83
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 83
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu His Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 84
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 84
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Asn Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 85
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 85
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Asn Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 86
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 86
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Gln Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 87
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 87
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Ile Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 88
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 88
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Val Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 89
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 89
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Gln Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 90
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 90
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu His Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 91
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 91
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Asn Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 92
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 92
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Ile Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 93
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 93
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Val Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 94
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 94
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Ile Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 95
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 95
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Val Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 96
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 96
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu His Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 97
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 97
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Gln Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 98
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 98
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Ile
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 99
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 99
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Val
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 100
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 100
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Ile Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 101
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 101
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Val Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 102
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 102
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Ile Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 103
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 103
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Val Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 104
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 104
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser His Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 105
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 105
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Ser Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 106
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 106
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Ile Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 107
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 107
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Val Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 108
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 108
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Gln Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 109
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 109
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu His Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 110
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 110
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Asn Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 111
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 111
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Ala Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 112
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 112
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Ser Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 113
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 113
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Ile Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 114
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 114
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Val Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 115
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 115
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Ile Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 116
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 116
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Val Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 117
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 117
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu His Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 118
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 118
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Gln Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 119
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 119
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Ile Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 120
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 120
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Val Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 121
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 121
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Ile Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 122
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 122
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Val Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 123
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 123
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val His Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 124
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 124
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Ile Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 125
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 125
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asn Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210> 126
<211> 191
<212> PRT
<213> Homo Sapiens
<400> 126
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Gln Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>127
<211>191
<212>PRT
<213>Homo Sapiens
<400>127
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val His Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>128
<211>191
<212>PRT
<213>Homo Sapiens
<400>128
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Ile Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>129
<211>191
<212>PRT
<213>Homo Sapiens
<400>129
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asn
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>130
<211>191
<212>PRT
<213>Homo Sapiens
<400>130
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Gln
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>131
<211>191
<212>PRT
<213>Homo Sapiens
<400>131
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Ile Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>132
<211>191
<212>PRT
<213>Homo Sapiens
<400>132
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Val Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>133
<211>191
<212>PRT
<213>Homo Sapiens
<400>133
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Ile Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>134
<211>191
<212>PRT
<213>Homo Sapiens
<400>134
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Val Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>135
<211>191
<212>PRT
<213>Homo Sapiens
<400>135
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Ash Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>136
<211>191
<212>PRT
<213>Homo Sapiens
<400>136
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Ash Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Gln Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>137
<211>191
<212>PRT
<213>Homo Sapiens
<400>137
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asn Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>138
<211>191
<212>PRT
<213>Homo Sapiens
<400>138
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Gln Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>139
<211>191
<212>PRT
<213>Homo Sapiens
<400>139
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Ile Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>140
<211>191
<212>PRT
<213>Homo Sapiens
<400>140
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Val Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>141
<211>191
<212>PRT
<213>Homo Sapiens
<400>141
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Gln Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>142
<211>191
<212>PRT
<213>Homo Sapiens
<400>142
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu His Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>143
<211>191
<212>PRT
<213>Homo Sapiens
<400>143
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Ash Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>144
<211>191
<212>PRT
<213>Homo Sapiens
<400>144
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Gln Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>145
<211>191
<212>PRT
<213>Homo Sapiens
<400>145
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu His Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>146
<211>191
<212>PRT
<213>Homo Sapiens
<400>146
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Asn Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>147
<211>191
<212>PRT
<213>Homo Sapiens
<400>147
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Ile Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>148
<211>191
<212>PRT
<213>Homo Sapiens
<400>148
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Val Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>149
<211>191
<212>PRT
<213>Homo Sapiens
<400>149
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Ile Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>150
<211>191
<212>PRT
<213>Homo Sapiens
<400>150
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Val Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>151
<211>191
<212>PRT
<213>Homo Sapiens
<400>151
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly His Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Ar9 Lys Asp Mer Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>152
<211>191
<212>PRT
<213>Homo Sapiens
<400>152
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Gln Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>153
<211>191
<212>PRT
<213>Homo Sapiens
<400>153
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Ile
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>154
<211>191
<212>PRT
<213>Homo Sapiens
<400>154
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Val
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>155
<211>191
<212>PRT
<213>Homo Sapiens
<400>155
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Gln Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>156
<211>191
<212>PRT
<213>Homo Sapiens
<400>156
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
His Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>157
<211>191
<212>PRT
<213>Homo Sapiens
<400>157
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Asn Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>158
<211>191
<212>PRT
<213>Homo Sapiens
<400>158
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asn Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>159
<211>191
<212>PRT
<213>Homo Sapiens
<400>159
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Gln Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>160
<211>191
<212>PRT
<213>Homo Sapiens
<400>160
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Ala Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>161
<211>191
<212>PRT
<213>Homo Sapiens
<400>161
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Ser Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>162
<211>191
<212>PRT
<213>Homo Sapiens
<400>162
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro His Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>163
<211>191
<212>PRT
<213>Homo Sapiens
<400>163
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Gln Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>164
<211>191
<212>PRT
<213>Homo Sapiens
<400>164
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Ile Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>165
<211>191
<212>PRT
<213>Homo Sapiens
<400>165
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Val Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>166
<211>191
<212>PRT
<213>Homo Sapiens
<400>166
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Asn Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>167
<211>191
<212>PRT
<213>Homo Sapiens
<400>167
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Gln Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>168
<211>191
<212>PRT
<213>Homo Sapiens
<400>168
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr His Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>169
<211>191
<212>PRT
<213>Homo Sapiens
<400>169
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Ile Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>170
<211>191
<212>PRT
<213>Homo Sapiens
<400>170
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Asn Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>171
<211>191
<212>PRT
<213>Homo Sapiens
<400>171
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Gln Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>172
<211>191
<212>PRT
<213>Homo Sapiens
<400>172
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Ile Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>173
<211>191
<212>PRT
<213>Homo Sapiens
<400>173
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Val Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>174
<211>191
<212>PRT
<213>Homo Sapiens
<400>174
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asn Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>175
<211>191
<212>PRT
<213>Homo Sapiens
<400>175
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Ash Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Gln Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>176
<211>191
<212>PRT
<213>Homo Sapiens
<400>176
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asn Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>177
<211>191
<212>PRT
<213>Homo Sapiens
<400>177
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Gln Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>178
<211>191
<212>PRT
<213>Homo Sapiens
<400>178
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asn Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>179
<211>191
<212>PRT
<213>Homo Sapiens
<400>179
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Gln Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>180
<211>191
<212>PRT
<213>Homo Sapiens
<400>180
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Ile Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>181
<211>191
<212>PRT
<213>Homo Sapiens
<400>181
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Val Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>182
<211>191
<212>PRT
<213>Homo Sapiens
<400>182
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala ASh Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Ile Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>183
<211>191
<212>PRT
<213>Homo Sapiens
<400>183
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Val Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>184
<211>191
<212>PRT
<213>Homo Sapiens
<400>184
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Asn Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>185
<211>191
<212>PRT
<213>Homo Sapiens
<400>185
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Gln Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>186
<211>191
<212>PRT
<213>Homo Sapiens
<400>186
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn His
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>187
<211>191
<212>PRT
<213>Homo Sapiens
<400>187
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Ash Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Ile
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>188
<211>191
<212>PRT
<213>Homo Sapiens
<400>188
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Ile Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>189
<211>191
<212>PRT
<213>Homo Sapiens
<400>189
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Val Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>190
<211>191
<212>PRT
<213>Homo Sapiens
<400>190
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Mer Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Ile Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>191
<211>191
<212>PRT
<213>Homo Sapiens
<400>191
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Val Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>192
<211>191
<212>PRT
<213>Homo Sapiens
<400>192
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu His Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>193
<211>191
<212>PRT
<213>Homo Sapiens
<400>193
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Ile Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>194
<211>191
<212>PRT
<213>Homo Sapiens
<400>194
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Ile Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>195
<211>191
<212>PRT
<213>Homo Sapiens
<400>195
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Val Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>196
<211>191
<212>PRT
<213>Homo Sapiens
<400>196
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe His Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>197
<211>191
<212>PRT
<213>Homo Sapiens
<400>197
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Gln Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>198
<211>191
<212>PRT
<213>Homo Sapiens
<400>198
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Asn Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>199
<211>191
<212>PRT
<213>Homo Sapiens
<400>199
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Gln Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>200
<211>191
<212>PRT
<213>Homo Sapiens
<400>200
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asn Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>201
<211>191
<212>PRT
<213>Homo Sapiens
<400>201
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Gln Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>202
<211>191
<212>PRT
<213>Homo Sapiens
<400>202
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Ile Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>203
<211>191
<212>PRT
<213>Homo Sapiens
<400>203
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Val Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>204
<211>191
<212>PRT
<213>Homo Sapiens
<400>204
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gin Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asn Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>205
<211>191
<212>PRT
<213>Homo Sapiens
<400>205
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Gln Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>206
<211>191
<212>PRT
<213>Homo Sapiens
<400>206
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Asn Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>207
<211>191
<212>PRT
<213>Homo Sapiens
<400>207
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Gln Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>208
<211>191
<212>PRT
<213>Homo Sapiens
<400>208
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Gln Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>209
<211>191
<212>PRT
<213>Homo Sapiens
<400>209
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val His Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>210
<211>191
<212>PRT
<213>Homo Sapiens
<400>210
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Asn Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>211
<211>191
<212>PRT
<213>Homo Sapiens
<400>211
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Ile
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>212
<211>191
<212>PRT
<213>Homo Sapiens
<400>212
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Val
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>213
<211>191
<212>PRT
<213>Homo Sapiens
<400>213
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Ile Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>214
<211>191
<212>PRT
<213>Homo Sapiens
<400>214
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Val Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>215
<211>191
<212>PRT
<213>Homo Sapiens
<400>215
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu His Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>216
<211>191
<212>PRT
<213>Homo Sapiens
<400>216
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Gln Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>217
<211>191
<212>PRT
<213>Homo Sapiens
<400>217
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys His Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>218
<211>191
<212>PRT
<213>Homo Sapiens
<400>218
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Gln Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>219
<211>191
<212>PRT
<213>Homo Sapiens
<400>219
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Gln Gly Ser Cys Gly Phe
180 185 190
<210>220
<211>191
<212>PRT
<213>Homo Sapiens
<400>220
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val His Gly Ser Cys Gly Phe
180 185 190
<210>221
<211>191
<212>PRT
<213>Homo Sapiens
<400>221
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Asn Gly Ser Cys Gly Phe
180 185 190
<210>222
<211>191
<212>PRT
<213>Homo Sapiens
<400>222
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Ile
180 185 190
<210>223
<211>191
<212>PRT
<213>Homo Sapiens
<400>223
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Val
180 185 190
<210>224
<211>191
<212>PRT
<213>Homo Sapiens
<400>224
Phe Pro Thr Ile Pro Glu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>225
<211>191
<212>PRT
<213>Homo Sapiens
<400>225
Phe Pro Thr Ile Pro Asp Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>226
<211>191
<212>PRT
<213>Homo Sapiens
<400>226
Phe Pro Thr Ile Pro Lys Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>227
<211>191
<212>PRT
<213>Homo Sapiens
<400>227
Phe Pro Thr Ile Pro Arg Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>228
<211>191
<212>PRT
<213>Homo Sapiens
<400>228
Phe Pro Thr Ile Pro Asn Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>229
<211>191
<212>PRT
<213>Homo Sapiens
<400>229
Phe Pro Thr Ile Pro Gln Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>230
<211>191
<212>PRT
<213>Homo Sapiens
<400>230
Phe Pro Thr Ile Pro Ser Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>231
<211>191
<212>PRT
<213>Homo Sapiens
<400>231
Phe Pro Thr Ile Pro Thr Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Cys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>232
<211>191
<212>PRT
<213>Homo Sapiens
<400>232
Phe Pro Thr Ile Pro Leu Ser Arg Glu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>233
<211>191
<212>PRT
<213>Homo Sapiens
<400>233
Phe Pro Thr Ile Pro Leu Ser Arg Asp Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>234
<211>191
<212>PRT
<213>Homo Sapiens
<400>234
Phe Pro Thr Ile Pro Leu Ser Arg Lys Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>235
<211>191
<212>PRT
<213>Homo Sapiens
<400>235
Phe Pro Thr Ile Pro Leu Ser Arg Arg Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>236
<211>191
<212>PRT
<213>Homo Sapiens
<400>236
Phe Pro Thr Ile Pro Leu Ser Arg Asn Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>237
<211>191
<212>PRT
<213>Homo Sapiens
<400>237
Phe Pro Thr Ile Pro Leu Ser Arg Gln Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>238
<211>191
<212>PRT
<213>Homo Sapiens
<400>238
Phe Pro Thr Ile Pro Leu Ser Arg Ser Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>239
<211>191
<212>PRT
<213>Homo Sapiens
<400>239
Phe Pro Thr Ile Pro Leu Ser Arg Thr Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>240
<211>191
<212>PRT
<213>Homo Sapiens
<400>240
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Glu Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>241
<211>191
<212>PRT
<213>Homo Sapiens
<400>241
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Asp Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>242
<211>191
<212>PRT
<213>Homo Sapiens
<400>242
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Lys Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Ar9 Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>243
<211>191
<212>PRT
<213>Homo Sapiens
<400>243
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Arg Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser ASn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>244
<211>191
<212>PRT
<213>Homo Sapiens
<400>244
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Asn Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>245
<211>191
<212>PRT
<213>Homo Sapiens
<400>245
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Gln Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>246
<211>191
<212>PRT
<213>Homo Sapiens
<400>246
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ser Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>247
<211>191
<212>PRT
<213>Homo Sapiens
<400>247
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Thr Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>248
<211>191
<212>PRT
<213>Homo Sapiens
<400>248
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Glu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>249
<211>191
<212>PRT
<213>Homo Sapiens
<400>249
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Asp Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>250
<211>191
<212>PRT
<213>Homo Sapiens
<400>250
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Lys Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>251
<211>191
<212>PRT
<213>Homo Sapiens
<400>251
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Arg Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>252
<211>191
<212>PRT
<213>Homo Sapiens
<400>252
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Asn Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>253
<211>191
<212>PRT
<213>Homo Sapiens
<400>253
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Gln Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>254
<211>191
<212>PRT
<213>Homo Sapiens
<400>254
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Ser Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>255
<211>191
<212>PRT
<213>Homo Sapiens
<400>255
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Thr Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>256
<211>191
<212>PRT
<213>Homo Sapiens
<400>256
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Glu His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>257
<211>191
<212>PRT
<213>Homo Sapiens
<400>257
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Asp His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>258
<211>191
<212>PRT
<213>Homo Sapiens
<400>258
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Lys His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>259
<211>191
<212>PRT
<213>Homo Sapiens
<400>259
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Arg His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>260
<211>191
<212>PRT
<213>Homo Sapiens
<400>260
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Asn His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>261
<211>191
<212>PRT
<213>Homo Sapiens
<400>261
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Gln His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>262
<211>191
<212>PRT
<213>Homo Sapiens
<400>262
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ser His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>263
<211>191
<212>PRT
<213>Homo Sapiens
<400>263
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Thr His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>264
<211>191
<212>PRT
<213>Homo Sapiens
<400>264
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Glu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>265
<211>191
<212>PRT
<213>Homo Sapiens
<400>265
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Asp His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>266
<211>191
<212>PRT
<213>Homo Sapiens
<400>266
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Lys His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>267
<211>191
<212>PRT
<213>Homo Sapiens
<400>267
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Arg His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>268
<211>191
<212>PRT
<213>Homo Sapiens
<400>268
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Asn His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>269
<211>191
<212>PRT
<213>Homo Sapiens
<400>269
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Gln His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>270
<211>191
<212>PRT
<213>Homo Sapiens
<400>270
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Ser His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>271
<211>191
<212>PRT
<213>Homo Sapiens
<400>271
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Thr His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>272
<211>191
<212>PRT
<213>Homo Sapiens
<400>272
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Glu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>273
<211>191
<212>PRT
<213>Homo Sapiens
<400>273
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Asp Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>274
<211>191
<212>PRT
<213>Homo Sapiens
<400>274
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Lys Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>275
<211>191
<212>PRT
<213>Homo Sapiens
<400>275
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Arg Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>276
<211>191
<212>PRT
<213>Homo Sapiens
<400>276
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Asn Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>277
<211>191
<212>PRT
<213>Homo Sapiens
<400>277
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Gln Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>278
<211>191
<212>PRT
<213>Homo Sapiens
<400>278
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Ser Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>279
<211>191
<212>PRT
<213>Homo Sapiens
<400>279
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Thr Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>280
<211>191
<212>PRT
<213>Homo Sapiens
<400>280
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Glu Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>281
<211>191
<212>PRT
<213>Homo Sapiens
<400>281
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Asp Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>282
<211>191
<212>PRT
<213>Homo Sapiens
<400>282
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Lys Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Mer Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>283
<211>191
<212>PRT
<213>Homo Sapiens
<400>283
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Arg Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>284
<211>191
<212>PRT
<213>Homo Sapiens
<400>284
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Asn Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>285
<211>191
<212>PRT
<213>Homo Sapiens
<400>285
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Gln Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>286
<211>191
<212>PRT
<213>Homo Sapiens
<400>286
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ser Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>287
<211>191
<212>PRT
<213>Homo Sapiens
<400>287
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Thr Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>288
<211>191
<212>PRT
<213>Homo Sapiens
<400>288
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Glu Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>289
<211>191
<212>PRT
<213>Homo Sapiens
<400>289
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Ash Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Ash Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Asp Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>290
<211>191
<212>PRT
<213>Homo Sapiens
<400>290
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Ash Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Lys Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>291
<211>191
<212>PRT
<213>Homo Sapiens
<400>291
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Arg Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>292
<211>191
<212>PRT
<213>Homo Sapiens
<400>292
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Asn Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>293
<211>191
<212>PRT
<213>Homo Sapiens
<400>293
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Gln Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>294
<211>191
<212>PRT
<213>Homo Sapiens
<400>294
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ser Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>295
<211>191
<212>PRT
<213>Homo Sapiens
<400>295
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Thr Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>296
<211>191
<212>PRT
<213>Homo Sapiens
<400>296
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Glu Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>297
<211>191
<212>PRT
<213>Homo Sapiens
<400>297
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Asp Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>298
<211>191
<212>PRT
<213>Homo Sapiens
<400>298
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Lys Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>299
<211>191
<212>PRT
<213>Homo Sapiens
<400>299
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Ash Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Arg Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>300
<211>191
<212>PRT
<213>Homo Sapiens
<400>300
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Asn Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>301
<211>191
<212>PRT
<213>Homo Sapiens
<400>301
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Gln Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>302
<211>191
<212>PRT
<213>Homo Sapiens
<400>302
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Ser Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>303
<211>191
<212>PRT
<213>Homo Sapiens
<400>303
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Thr Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>304
<211>191
<212>PRT
<213>Homo Sapiens
<400>304
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Glu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>305
<211>191
<212>PRT
<213>Homo Sapiens
<400>305
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Asp Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>306
<211>191
<212>PRT
<213>Homo Sapiens
<400>306
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Lys Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>307
<211>191
<212>PRT
<213>Homo Sapiens
<400>307
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Arg Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>308
<211>191
<212>PRT
<213>Homo Sapiens
<400>308
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Asn Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>309
<211>191
<212>PRT
<213>Homo Sapiens
<400>309
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Gln Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>310
<211>191
<212>PRT
<213>Homo Sapiens
<400>310
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Ser Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>311
<211>191
<212>PRT
<213>Homo Sapiens
<400>311
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Thr Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>312
<211>191
<212>PRT
<213>Homo Sapiens
<400>312
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Glu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>313
<211>191
<212>PRT
<213>Homo Sapiens
<400>313
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Asp Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>314
<211>191
<212>PRT
<213>Homo Sapiens
<400>314
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Lys Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>315
<211>191
<212>PRT
<213>Homo Sapiens
<400>315
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Arg Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>316
<211>191
<212>PRT
<213>Homo Sapiens
<400>316
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Asn Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>317
<211>191
<212>PRT
<213>Homo Sapiens
<400>317
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Gln Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>318
<211>191
<212>PRT
<213>Homo Sapiens
<400>318
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Ser Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>319
<211>191
<212>PRT
<213>Homo Sapiens
<400>319
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Thr Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>320
<211>191
<212>PRT
<213>Homo Sapiens
<400>320
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Glu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>321
<211>191
<212>PRT
<213>Homo Sapiens
<400>321
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Asp Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>322
<211>191
<212>PRT
<213>Homo Sapiens
<400>322
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Lys Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>323
<211>191
<212>PRT
<213>Homo Sapiens
<400>323
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Arg Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>324
<211>191
<212>PRT
<213>Homo Sapiens
<400>324
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Asn Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>325
<211>191
<212>PRT
<213>Homo Sapiens
<400>325
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Gln Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>326
<211>191
<212>PRT
<213>Homo Sapiens
<400>326
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Ser Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>327
<211>191
<212>PRT
<213>Homo Sapiens
<400>327
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Thr Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>328
<211>191
<212>PRT
<213>Homo Sapiens
<400>328
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Glu Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>329
<211>191
<212>PRT
<213>Homo Sapiens
<400>329
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Asp Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>330
<211>191
<212>PRT
<213>Homo Sapiens
<400>330
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Lys Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>331
<211>191
<212>PRT
<213>Homo Sapiens
<400>331
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Arg Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>332
<211>191
<212>PRT
<213>Homo Sapiens
<400>332
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Asn Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>333
<211>191
<212>PRT
<213>Homo Sapiens
<400>333
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Gln Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>334
<211>191
<212>PRT
<213>Homo Sapiens
<400>334
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ser Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>335
<211>191
<212>PRT
<213>Homo Sapiens
<400>335
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Thr Gln Thr Leu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>336
<211>191
<212>PRT
<213>Homo Sapiens
<400>336
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Glu Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>337
<211>191
<212>PRT
<213>Homo Sapiens
<400>337
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Asp Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>338
<211>191
<212>PRT
<213>Homo Sapiens
<400>338
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Lys Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>339
<211>191
<212>PRT
<213>Homo Sapiens
<400>339
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Arg Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>340
<211>191
<212>PRT
<213>Homo Sapiens
<400>340
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Asn Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>341
<211>191
<212>PRT
<213>Homo Sapiens
<400>341
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Gln Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>342
<211>191
<212>PRT
<213>Homo Sapiens
<400>342
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Ser Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>343
<211>191
<212>PRT
<213>Homo Sapiens
<400>343
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Thr Met Gly Arg Leu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>344
<211>191
<212>PRT
<213>Homo Sapiens
<400>344
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Glu
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>345
<211>191
<212>PRT
<213>Homo Sapiens
<400>345
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Asp
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>346
<211>191
<212>PRT
<213>Homo Sapiens
<400>346
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Lys
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>347
<211>191
<212>PRT
<213>Homo Sapiens
<400>347
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Arg
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>348
<211>191
<212>PRT
<213>Homo Sapiens
<400>348
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Asn
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>349
<211>191
<212>PRT
<213>Homo Sapiens
<400>349
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Gln
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>350
<211>191
<212>PRT
<213>Homo Sapiens
<400>350
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Ser
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>351
<211>191
<212>PRT
<213>Homo Sapiens
<400>351
Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg
1 5 10 15
Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu
20 25 30
Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro
35 40 45
Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg
50 55 60
Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu
65 70 75 80
Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val
85 90 95
Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp
100 105 110
Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Thr
115 120 125
Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser
130 135 140
Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr
145 150 155 160
Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe
165 170 175
Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
180 185 190
<210>352
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>352
gggcagtgcc atcgccacca ttcccttatc caggcgcctg gataagggaa tggtggcgat 60
ggcactgccc 70
<210>353
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>353
gggcagtgcc atcagcacca ttcccttatc caggcgcctg gataagggaa tggtgctgat 60
ggcactgccc 70
<210>354
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>354
gggcagtgcc gtggccacca ttcccttatc caggcgcctg gataagggaa tggtggccac 60
ggcactgccc 70
<210>355
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>355
gggcagtgcc gtgagcacca ttcccttatc caggcgcctg gataagggaa tggtgctcac 60
ggcactgccc 70
<210>356
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>356
gggcagtgcc ttcccaacca ttgccttatc caggcgcctg gataaggcaa tggttgggaa 60
ggcactgccc 70
<210>357
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>357
gggcagtgcc ttcccaacca ttagcttatc caggcgcctg gataagctaa tggttgggaa 60
ggcactgccc 70
<210>358
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>358
ggcagtgcct tcccaaccat tcccatctcc aggctttttg caaaaagcct ggagatggga 60
atggttggga aggcactgcc 80
<210>359
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>359
ggcagtgcct tcccaaccat tcccgtgtcc aggctttttg caaaaagcct ggacacggga 60
atggttggga aggcactgcc 80
<210>360
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>360
cattccctta tcccaccttt ttgacaacgc tatgctccgc gcggagcata gcgttgtcaa 60
aaaggtggga taagggaatg 80
<210>361
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>361
cattccctta tcccagcttt ttgacaacgc tatgctccgc gcggagcata gcgttgtcaa 60
aaagctggga taagggaatg 80
<210>362
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>362
cccttatcca ggatctttga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tcaaagatcc tggataaggg 80
<210>363
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>363
cccttatcca gggtgtttga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tcaaacaccc tggataaggg 80
<210>364
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>364
cccttatcca ggcttatcga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tcgataagcc tggataaggg 80
<210>365
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>365
cccttatcca ggcttgtgga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tccacaagcc tggataaggg 80
<210>366
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>366
ccaggctttt taacaacgct atgctccgcg cccgggcgcg gagcatagcg ttgttaaaaa 60
gcctgg 66
<210>367
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>367
ccaggctttt tcagaacgct atgctccgcg cccgggcgcg gagcatagcg ttctgaaaaa 60
gcctgg 66
<210>368
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>368
ccaggctttt tgacaacgct atcctccgcg cccgggcgcg gaggatagcg ttgtcaaaaa 60
gcctgg 66
<210>369
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>369
ccaggctttt tgacaacgct gtgctccgcg cccgggcgcg gagcacagcg ttgtcaaaaa 60
gcctgg 66
<210>370
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>370
caacgctatg atccgcgccc atcgtctgca ccagctggcg ccagctggtg cagacgatgg 60
gcgcggatca tagcgttg 78
<210>371
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>371
caacgctatg gtgcgcgccc atcgtctgca ccagctggcg ccagctggtg cagacgatgg 60
gcgcgcacca tagcgttg 78
<210>372
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>372
cgctatgctc cacgcccatc gtctgcacca gctggccggc cagctggtgc agacgatggg 60
cgtggagcat agcg 74
<210>373
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>373
cgctatgctc caggcccatc gtctgcacca gctggccggc cagctggtgc agacgatggg 60
cctggagcat agcg 74
<210>374
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>374
gctccgcgcc catcacctgc accagctggc cggccagctg gtgcaggtga tgggcgcgga 60
gc 62
<210>375
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>375
gctccgcgcc catcagctgc accagctggc cggccagctg gtgcagctga tgggcgcgga 60
gc 62
<210>376
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>376
gctccgcgcc catcgtatcc accagctggc cggccagctg gtggatacga tgggcgcgga 60
gc 62
<210>377
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>377
gctccgcgcc catcgtgtgc accagctggc cggccagctg gtgcacacga tgggcgcgga 60
gc 62
<210>378
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>378
gctccgcgcc catcgtctgc accagatcgc ctttgacacc ggtgtcaaag gcgatctggt 60
gcagacgatg ggcgcggagc 80
<210>379
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>379
ccgcgcccat cgtctgcacc aggtggcctt tgacaccggt gtcaaaggcc acctggtgca 60
gacgatgggc gcgg 74
<210>380
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>380
gcgcccatcg tctgcaccag ctggccatcg acacctaccg gtaggtgtcg atggccagct 60
ggtgcagacg atgggcgc 78
<210>381
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>381
gcgcccatcg tctgcaccag ctggccgtgg acacctaccg gtaggtgtcc acggccagct 60
ggtgcagacg atgggcgc 78
<210>382
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>382
cccatcgtct gcaccagctg gcctttaaca cctaccaggc ctggtaggtg ttaaaggcca 60
gctggtgcag acgatggg 78
<210>383
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>383
cgtctgcacc agctggcctt tcagacctac caggcctggt aggtctgaaa ggccagctgg 60
tgcagacg 68
<210>384
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>384
gcaccagctg gcctttgaca cccaccagga gtttgcaaac tcctggtggg tgtcaaaggc 60
cagctggtgc 70
<210>385
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>385
gcaccagctg gcctttgaca ccatccagga gtttgcaaac tcctggatgg tgtcaaaggc 60
cagctggtgc 70
<210>386
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>386
ccagctggcc tttgacacct accagcagtt tgaagaagcc ggcttcttca aactgctggt 60
aggtgtcaaa ggccagctgg 80
<210>387
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>387
ccagctggcc tttgacacct accagcactt tgaagaagcc ggcttcttca aagtgctggt 60
aggtgtcaaa ggccagctgg 80
<210>388
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>388
ccagctggcc tttgacacct accagaactt tgaagaagcc ggcttcttca aagttctggt 60
aggtgtcaaa ggccagctgg 80
<210>389
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>389
cctaccagga gatcgaagaa gcctatatcc cgggatatag gcttcttcga tctcctggta 60
gg 62
<210>390
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>390
cctaccagga ggtggaagaa gcctatatcc cgggatatag gcttcttcca cctcctggta 60
gg 62
<210>391
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>391
cctaccagga gtttcaggaa gcctatatcc cgggatatag gcttcctgaa actcctggta 60
gg 62
<210>392
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>392
cctaccagga gtttcacgaa gcctatatcc cgggatatag gcttcgtgaa actcctggta 60
gg 62
<210>393
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>393
gacacctacc aggagtttaa cgaagcctat atcccgggat ataggcttcg ttaaactcct 60
ggtaggtgtc 70
<210>394
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>394
cctaccagga gtttgaacag gcctatatcc cgggatatag gcctgttcaa actcctggta 60
gg 62
<210>395
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>395
cctaccagga gtttgaacac gcctatatcc cgggatatag gcgtgttcaa actcctggta 60
gg 62
<210>396
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>396
gacacctacc aggagtttga aaacgcctat atcccgggat ataggcgttt tcaaactcct 60
ggtaggtgtc 70
<210>397
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>397
ccaggagttt gaagaagccc acatcccaaa ggcctttggg atgtgggctt cttcaaactc 60
ctgg 64
<210>398
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>398
ccaggagttt gaagaagcca tcatcccaaa ggcctttggg atgatggctt cttcaaactc 60
ctgg 64
<210>399
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>399
ccaggagttt gaagaagcct atatcgccaa ggaacagaag cttctgttcc ttggcgatat 60
aggcttcttc aaactcctgg 80
<210>400
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>400
ccaggagttt gaagaagcct atatcagcaa ggaacagaag cttctgttcc ttgctgatat 60
aggcttcttc aaactcctgg 80
<210>401
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>401
gcctatatcc caaacgaaca gaagtattca ttcctgcagc tgcaggaatg aatacttctg 60
ttcgtttggg atataggc 78
<210>402
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>402
gcctatatcc cacaggaaca gaagtattca ttcctgcagc tgcaggaatg aatacttctg 60
ttcctgtggg atataggc 78
<210>403
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>403
gcctatatcc caaagcagca gaagtattca ttcctgcagc tgcaggaatg aatacttctg 60
ctgctttggg atataggc 78
<210>404
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>404
gcctatatcc caaagcacca gaagtattca ttcctgcagc tgcaggaatg aatacttctg 60
gtgctttggg atataggc 78
<210>405
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>405
gcctatatcc caaagaacca gaagtattca ttcctgcagc tgcaggaatg aatacttctg 60
gttctttggg atataggc 78
<210>406
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>406
cccaaaggaa cagaactatt cattcctgca gaacccccgg gggttctgca ggaatgaata 60
gttctgttcc tttggg 76
<210>407
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>407
cccaaaggaa cagcagtatt cattcctgca gaacccccgg gggttctgca ggaatgaata 60
ctgctgttcc tttggg 76
<210>408
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>408
ggaacagaag cactcattcc tgcagaaccc ccagaccggt ctgggggttc tgcaggaatg 60
agtgcttctg ttcc 74
<210>409
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>409
ggaacagaag atctcattcc tgcagaaccc ccagaccggt ctgggggttc tgcaggaatg 60
agatcttctg ttcc 74
<210>410
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>410
gaagtattca atcctgcaga acccccagac ctcccgggag gtctgggggt tctgcaggat 60
tgaatacttc 70
<210>411
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>411
gaagtattca gtgctgcaga acccccagac ctcccgggag gtctgggggt tctgcagcac 60
tgaatacttc 70
<210>412
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>412
gtattcattc atccagaacc cccagacctc ccgggaggtc tgggggttct ggatgaatga 60
atac 64
<210>413
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>413
gtattcattc gtgcagaacc cccagacctc ccgggaggtc tgggggttct gcacgaatga 60
atac 64
<210>414
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>414
cctgcagaac gcccagacct ccctctgttt cgaaacagag ggaggtctgg gcgttctgca 60
gg 62
<210>415
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>415
cctgcagaac agccagacct ccctctgttt cgaaacagag ggaggtctgg ctgttctgca 60
gg 62
<210>416
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>416
cctgcagaac ccccagacct ccatctgttt ctcagctgag aaacagatgg aggtctgggg 60
gttctgcagg 70
<210>417
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>417
cccccagacc tccgtgtgtt tctcagagtc gactctgaga aacacacgga ggtctggggg 60
<210>418
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>418
cccccagacc tccctctgta tctcagagtc tattccgcgg aatagactct gagatacaga 60
gggaggtctg gggg 74
<210>419
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>419
cccccagacc tccctctgtg tgtcagagtc tattccgcgg aatagactct gacacacaga 60
gggaggtctg gggg 74
<210>420
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>420
cccccagacc tccctctgtt tctcacagtc tattccgacg tcggaataga ctgtgagaaa 60
cagagggagg tctggggg 78
<210>421
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>421
cccccagacc tccctctgtt tctcacactc tattccgacg tcggaataga gtgtgagaaa 60
cagagggagg tctggggg 78
<210>422
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>422
cccccagacc tccctctgtt tctcaaactc tattccgacg tcggaataga gtttgagaaa 60
cagagggagg tctggggg 78
<210>423
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>423
cagagtctat tgccacaccc tccaacaggg aggcctccct gttggagggt gtggcaatag 60
actctg 66
<210>424
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>424
cagagtctat tagcacaccc tccaacaggg aggcctccct gttggagggt gtgctaatag 60
actctg 66
<210>425
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>425
gagtctattc cgacagcctc caacagggag gcctccctgt tggaggctgt cggaatagac 60
tc 62
<210>426
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>426
gagtctattc cgacaagctc caacagggag gcctccctgt tggagcttgt cggaatagac 60
tc 62
<210>427
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>427
ccgacaccct ccaaccacga ggaaacacaa cgttgtgttt cctcgtggtt ggagggtgtc 60
gg 62
<210>428
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>428
ccgacaccct ccaaccagga ggaaacacaa cgttgtgttt cctcctggtt ggagggtgtc 60
gg 62
<210>429
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>429
ccgacaccct ccaacaggca ggaaacacaa cgttgtgttt cctgcctgtt ggagggtgtc 60
gg 62
<210>430
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>430
ccgacaccct ccaacaggca cgaaacacaa cgttgtgttt cgtgcctgtt ggagggtgtc 60
gg 62
<210>431
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>431
ccgacaccct ccaacaggaa cgaaacacaa cgttgtgttt cgttcctgtt ggagggtgtc 60
gg 62
<210>432
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>432
ccgacaccct ccaacaggga gcagacacaa cagaaatccg gatttctgtt gtgtctgctc 60
cctgttggag ggtgtcgg 78
<210>433
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>433
ccgacaccct ccaacaggga gcacacacaa cagaaatccg gatttctgtt gtgtgtgctc 60
cctgttggag ggtgtcgg 78
<210>434
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>434
ccgacaccct ccaacaggga gaacacacaa cagaaatccg gatttctgtt gtgtgttctc 60
cctgttggag ggtgtcgg 78
<210>435
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>435
ggaggaaaca caacagaact ccaacctaga gctgctccgc gcggagcagc tctaggttgg 60
agttctgttg tgtttcctcc 80
<210>436
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>436
ggaggaaaca caacagcagt ccaacctaga gctgctccgc gcggagcagc tctaggttgg 60
actgctgttg tgtttcctcc 80
<210>437
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>437
gaaatccaac atcgagctgc tccgcatctc cctgctgcgc agcagggaga tgcggagcag 60
ctcgatgttg gatttc 76
<210>438
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>438
gaaatccaac gtggagctgc tccgcatctc cctgctgcgc agcagggaga tgcggagcag 60
ctccacgttg gatttc 76
<210>439
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>439
gaaatccaac ctacagctgc tccgcatctc cctgctgcgc agcagggaga tgcggagcag 60
ctgtaggttg gatttc 76
<210>440
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>440
gaaatccaac ctacacctgc tccgcatctc cctgctgcgc agcagggaga tgcggagcag 60
gtgtaggttg gatttc 76
<210>441
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>441
gaaatccaac ctaaacctgc tccgcatctc cctgctgcgc agcagggaga tgcggagcag 60
gtttaggttg gatttc 76
<210>442
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>442
ccaacctaga gatcctccgc atctccctgc tgcgcagcag ggagatgcgg aggatctcta 60
ggttgg 66
<210>443
<211>65
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>443
ccaacctaga ggtgctccgc atctccctgc tgcgagcagg gagatgcgga gcacctctag 60
gttgg 65
<210>444
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>444
cctagagctg atccgcatct ccctgctgct cgagcagcag ggagatgcgg atcagctcta 60
gg 62
<210>445
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>445
cctagagctg gtgcgcatct ccctgctgct cgagcagcag ggagatgcgc accagctcta 60
gg 62
<210>446
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>446
cctagagctg ctccacatct ccctgctgct cgagcagcag ggagatgtgg agcagctcta 60
gg 62
<210>447
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>447
cctagagctg ctccagatct ccctgctgct cgagcagcag ggagatctgg agcagctcta 60
gg 62
<210>448
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>448
ccgcatctcc atcctgctca tccagtcgtg gctggagccg gctccagcca cgactggatg 60
agcaggatgg agatgcgg 78
<210>449
<211>77
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>449
ccgcatctcc gtgctgctca tccagtcgtg gctggagccg ctccagccac gactggatga 60
gcagcacgga gatgcgg 77
<210>450
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>450
cgcatctccc tgatcctcat ccagtcgtgg ctggagcccg cgggctccag ccacgactgg 60
atgaggatca gggagatgcg 80
<210>451
<211>79
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>451
cgcatctccc tggtgctcat ccagtcgtgg ctggagcccg gggctccagc cacgactgga 60
tgagcaccag ggagatgcg 79
<210>452
<211>75
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>452
ctccctgctg atcatccagt cgtggctgga gcccgtgcca cgggctccag ccacgactgg 60
atgatcagca gggag 75
<210>453
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>453
ctccctgctg gtgatccagt cgtggctgga gcccgcgggc tccagccacg actggatcac 60
cagcagggag 70
<210>454
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>454
ccctgctgct catccagtcg cacctggagc ccgcgggctc caggtgcgac tggatgagca 60
gcaggg 66
<210>455
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>455
ccctgctgct catccagtcg agcctggagc ccgcgggctc caggctcgac tggatgagca 60
gcaggg 66
<210>456
<211>72
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>456
ccctgctgct catccagtcg tggatcgagc ccgtgcgcac gggctcgatc cacgactgga 60
tgagcagcag gg 72
<210>457
<211>72
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>457
ccctgctgct catccagtcg tgggtggagc ccgtgcgcac gggctccacc cacgactgga 60
tgagcagcag gg 72
<210>458
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>458
gtcgtggctg cagcccgtgc agttcctcag gcctgaggaa ctgcacgggc tgcagccacg 60
ac 62
<210>459
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>459
gtcgtggctg caccccgtgc agttcctcag gcctgaggaa ctgcacgggg tgcagccacg 60
ac 62
<210>460
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>460
ccagtcgtgg ctgaaccccg tgcagttcct caggcctgag gaactgcacg gggttcagcc 60
acgactgg 68
<210>461
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>461
cgtggctgga ggccgtgcag ttcctcagga gctcctgagg aactgcacgg cctccagcca 60
cg 62
<210>462
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>462
ccagtcgtgg ctggagagcg tgcagttcct caggcctgag gaactgcacg ctctccagcc 60
acgactgg 68
<210>463
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>463
ggctggagcc cgtgcagatc ctcaggagtg cactcctgag gatctgcacg ggctccagcc 60
<210>464
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>464
ggctggagcc cgtgcaggtg ctcaggagtg cactcctgag cacctgcacg ggctccagcc 60
<210>465
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>465
ggctggagcc cgtgcagttc atcaggagtg tcttcgccgg cgaagacact cctgatgaac 60
tgcacgggct ccagcc 76
<210>466
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>466
ggctggagcc cgtgcagttc gtgaggagtg tcttcgccgg cgaagacact cctcacgaac 60
tgcacgggct ccagcc 76
<210>467
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>467
ggctggagcc cgtgcagttc ctccacagtg tcttcgccgg cgaagacact gtggaggaac 60
tgcacgggct ccagcc 76
<210>468
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>468
ggctggagcc cgtgcagttc ctccagagtg tcttcgccgg cgaagacact ctggaggaac 60
tgcacgggct ccagcc 76
<210>469
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>469
cctcaggagt gtcatcgcca acagcctggt gtacggcgcc ggcgccgtac accaggctgt 60
tggcgatgac actcctgagg 80
<210>470
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>470
caggagtgtc gtggccaaca gcctggtgta cggcgccggc gccgtacacc aggctgttgg 60
ccacgacact cctg 74
<210>471
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>471
cgccaacagc atcgtgtacg gcgcctctga cagcgctgtc agaggcgccg tacacgatgc 60
tgttggcg 68
<210>472
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>472
cgccaacagc gtggtgtacg gcgcctctga cgtcagaggc gccgtacacc acgctgttgg 60
cg 62
<210>473
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>473
cgccaacagc ctggtgcacg gcgcctctga cgtcagaggc gccgtgcacc aggctgttgg 60
cg 62
<210>474
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>474
cgccaacagc ctggtgatcg gcgcctctga cgtcagaggc gccgatcacc aggctgttgg 60
cg 62
<210>475
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>475
cggcgcctct aacagcaacg tctatgacct ccggaggtca tagacgttgc tgttagaggc 60
gccg 64
<210>476
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>476
cggcgcctct cagagcaacg tctatgacct ccggaggtca tagacgttgc tctgagaggc 60
gccg 64
<210>477
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>477
cggcgcctct gacagcaacg tccacgacct cctaaaggcc tttaggaggt cgtggacgtt 60
gctgtcagag gcgccg 76
<210>478
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>478
cggcgcctct gacagcaacg tcatcgacct cctaaaggcc tttaggaggt cgatgacgtt 60
gctgtcagag gcgccg 76
<210>479
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>479
cggcgcctct gacagcaacg tctataacct cctaaaggcc tttaggaggt tatagacgtt 60
gctgtcagag gcgccg 76
<210>480
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>480
cggcgcctct gacagcaacg tctatcagct cctaaaggcc tttaggagct gatagacgtt 60
gctgtcagag gcgccg 76
<210>481
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>481
gcgcctctga cagcaacgtc tatgacatcc taaaggaccg gtcctttagg atgtcataga 60
cgttgctgtc agaggcgc 78
<210>482
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>482
gcgcctctga cagcaacgtc tatgacgtgc taaaggaccg gtcctttagc acgtcataga 60
cgttgctgtc agaggcgc 78
<210>483
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>483
cgtctatgac ctcatcaagg acctagagga aggcgccttc ctctaggtcc ttgatgaggt 60
catagacg 68
<210>484
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>484
cgtctatgac ctcgtgaagg acctagagga aggcgccttc ctctaggtcc ttcacgaggt 60
catagacg 68
<210>485
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>485
cgtctatgac ctcctaaacg acctagagga aggcgccttc ctctaggtcg tttaggaggt 60
catagacg 68
<210>486
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>486
cgtctatgac ctcctacagg acctagagga aggcgccttc ctctaggtcc tgtaggaggt 60
catagacg 68
<210>487
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>487
gacctcctaa agaacctaga ggaaggcatc cggatgcctt cctctaggtt ctttaggagg 60
tc 62
<210>488
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>488
gacctcctaa agcagctaga ggaaggcatc cggatgcctt cctctagctg ctttaggagg 60
tc 62
<210>489
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>489
gacctcctaa aggacatcga ggaaggcatc cggatgcctt cctcgatgtc ctttaggagg 60
tc 62
<210>490
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>490
gacctcctaa aggacgtgga ggaaggcatc cggatgcctt cctccacgtc ctttaggagg 60
tc 62
<210>491
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>491
cctaaaggac ctacaggaag gcatccaaac gctgatgggg ccccatcagc gtttggatgc 60
cttcctgtag gtcctttagg 80
<210>492
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>492
cctaaaggac ctacacgaag gcatccaaac gctgatgggg ccccatcagc gtttggatgc 60
cttcgtgtag gtcctttagg 80
<210>493
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>493
cctaaaggac ctaaacgaag gcatccaaac gctgatgggg ccccatcagc gtttggatgc 60
cttcgtttag gtcctttagg 80
<210>494
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>494
ggacctagag cagggcatcc aaacgctgat ggggaggcgc ctccccatca gcgtttggat 60
gccctgctct aggtcc 76
<210>495
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>495
ggacctagag cacggcatcc aaacgctgat ggggaggcgc ctccccatca gcgtttggat 60
gccgtgctct aggtcc 76
<210>496
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>496
ggacctagag aacggcatcc aaacgctgat ggggaggcgc ctccccatca gcgtttggat 60
gccgttctct aggtcc 76
<210>497
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>497
gcatccaaac gatcatgggg aggctggaag atggcagccc gggctgccat cttccagcct 60
ccccatgatc gtttggatgc 80
<210>498
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>498
gcatccaaac ggtgatgggg aggctggaag atggcagccc gggctgccat cttccagcct 60
ccccatcacc gtttggatgc 80
<210>499
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>499
ccaaacgctg atcgggaggc tggaagatgg cagcccccgc gggggctgcc atcttccagc 60
ctcccgatca gcgtttgg 78
<210>500
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>500
ccaaacgctg gtggggaggc tggaagatgg cagcccccgc gggggctgcc atcttccagc 60
ctccccacca gcgtttgg 78
<210>501
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>501
cgctgatggg gcacctggaa gatggcagcc cccggccggg ggctgccatc ttccaggtgc 60
cccatcagcg 70
<210>502
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>502
cgctgatggg gcagctggaa gatggcagcc cccggccggg ggctgccatc ttccagctgc 60
cccatcagcg 70
<210>503
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>503
cgctgatggg gaggatcgaa gatggcagcc cccggccggg ggctgccatc ttcgatcctc 60
cccatcagcg 70
<210>504
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>504
cgctgatggg gagggtggaa gatggcagcc cccggccggg ggctgccatc ttccaccctc 60
cccatcagcg 70
<210>505
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>505
ggggaggctg caggatggca gcccccggac tgggcgccca gtccgggggc tgccatcctg 60
cagcctcccc 70
<210>506
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>506
ggggaggctg cacgatggca gcccccggac tgggcgccca gtccgggggc tgccatcgtg 60
cagcctcccc 70
<210>507
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>507
ggggaggctg aacgatggca gcccccggac tgggcgccca gtccgggggc tgccatcgtt 60
cagcctcccc 70
<210>508
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>508
ggggaggctg gaaaacggca gcccccggac tgggcgccca gtccgggggc tgccgttttc 60
cagcctcccc 70
<210>509
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>509
ggggaggctg gaacagggca gcccccggac tgggcgccca gtccgggggc tgccctgttc 60
cagcctcccc 70
<210>510
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>510
ggggaggctg gaagatggca gcgcccggac tgggcgccca gtccgggcgc tgccatcttc 60
cagcctcccc 70
<210>511
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>511
ggggaggctg gaagatggca gcagccggac tgggcgccca gtccggctgc tgccatcttc 60
cagcctcccc 70
<210>512
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>512
ggggaggctg gaagatggca gcccccacac tgggcagatc gatctgccca gtgtgggggc 60
tgccatcttc cagcctcccc 80
<210>513
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>513
ggggaggctg gaagatggca gcccccagac tgggcagatc gatctgccca gtctgggggc 60
tgccatcttc cagcctcccc 80
<210>514
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>514
ggcagccccc ggactgggca gatcatcaag cagacctacg taggtctgct tgatgatctg 60
cccagtccgg gggctgcc 78
<210>515
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>515
ggcagccccc ggactgggca gatcgtgaag cagacctacg taggtctgct tcacgatctg 60
cccagtccgg gggctgcc 78
<210>516
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>516
gcccccggac tgggcagatc ttcaaccaga cctacagcgc tgtaggtctg gttgaagatc 60
tgcccagtcc gggggc 76
<210>517
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>517
gcccccggac tgggcagatc ttccagcaga cctacagcgc tgtaggtctg ctggaagatc 60
tgcccagtcc gggggc 76
<210>518
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>518
ggactgggca gatcttcaag cagacccaca gcaagttcgc gaacttgctg tgggtctgct 60
tgaagatctg cccagtcc 78
<210>519
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>519
ggactgggca gatcttcaag cagaccatca gcaagttcgc gaacttgctg atggtctgct 60
tgaagatctg cccagtcc 78
<210>520
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>520
gcagacctac agcaacttcg acacaaactc gagtttgtgt cgaagttgct gtaggtctgc 60
<210>521
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>521
gcagacctac agccagttcg acacaaactc gagtttgtgt cgaactggct gtaggtctgc 60
<210>522
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>522
cctacagcaa gatcgacaca aactcacaca acgatgacgc gcgtcatcgt tgtgtgagtt 60
tgtgtcgatc ttgctgtagg 80
<210>523
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>523
gcagacctac agcaaggtgg acacaaactc gagtttgtgt ccaccttgct gtaggtctgc 60
<210>524
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>524
cctacagcaa gttcaacaca aactcacaca acgatgacgc gcgtcatcgt tgtgtgagtt 60
tgtgttgaac ttgctgtagg 80
<210>525
<211>72
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>525
cagcaagttc cagacaaact cacacaacga tgacgcgcgt catcgttgtg tgagtttgtc 60
tggaacttgc tg 72
<210>526
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>526
ctcacacaac aacgacgcac tactcaagaa ctacgggcgc ccgtagttct tgagtagtgc 60
gtcgttgttg tgtgag 76
<210>527
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>527
ctcacacaac caggacgcac tactcaagaa ctacgggcgc ccgtagttct tgagtagtgc 60
gtcctggttg tgtgag 76
<210>528
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>528
cacacaacga taacgcacta ctcaagaact acgggctgcg cagcccgtag ttcttgagta 60
gtgcgttatc gttgtgtg 78
<210>529
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>529
cacacaacga tcaggcacta ctcaagaact acgggctgcg cagcccgtag ttcttgagta 60
gtgcctgatc gttgtgtg 78
<210>530
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>530
cgatgacgca atcctcaaga actacgggct gcgcagcccg tagttcttga ggattgcgtc 60
atcg 64
<210>531
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>531
cgatgacgca gtgctcaaga actacgggct gcgcagcccg tagttcttga gcactgcgtc 60
atcg 64
<210>532
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>532
cgatgacgca ctaatcaaga actacgggct gcgcagcccg tagttcttga ttagtgcgtc 60
atcg 64
<210>533
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>533
cgatgacgca ctagtgaaga actacgggct gcgcagcccg tagttcttca ctagtgcgtc 60
atcg 64
<210>534
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>534
gacgcactac tcaacaacta cgggctgctc gagcagcccg tagttgttga gtagtgcgtc 60
<210>535
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>535
gacgcactac tccagaacta cgggctgctc gagcagcccg tagttctgga gtagtgcgtc 60
<210>536
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>536
cgcactactc aagaaccacg ggctgctcta ctgcgcagta gagcagcccg tggttcttga 60
gtagtgcg 68
<210>537
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>537
cgcactactc aagaacatcg ggctgctcta ctgcgcagta gagcagcccg atgttcttga 60
gtagtgcg 68
<210>538
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>538
gaactacggg atcctctact gcttcaggaa ggccttcctg aagcagtaga ggatcccgta 60
gttc 64
<210>539
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>539
gaactacggg gtgctctact gcttcaggaa ggccttcctg aagcagtaga gcaccccgta 60
gttc 64
<210>540
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>540
ctacgggctg atctactgct tcaggaagga cgtccttcct gaagcagtag atcagcccgt 60
ag 62
<210>541
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>541
ctacgggctg gtgtactgct tcaggaagga cgtccttcct gaagcagtac accagcccgt 60
ag 62
<210>542
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>542
cgggctgctc cactgcttca ggaaggacat ggccatgtcc ttcctgaagc agtggagcag 60
cccg 64
<210>543
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>543
cgggctgctc atctgcttca ggaaggacat ggccatgtcc ttcctgaagc agatgagcag 60
cccg 64
<210>544
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>544
cgggctgctc tactgcatca ggaaggacat ggccatgtcc ttcctgatgc agtagagcag 60
cccg 64
<210>545
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>545
cgggctgctc tactgcgtga ggaaggacat ggccatgtcc ttcctcacgc agtagagcag 60
cccg 64
<210>546
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>546
cgggctgctc tactgcttcc acaaggacat ggccatgtcc ttgtggaagc agtagagcag 60
cccg 64
<210>547
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>547
cgggctgctc tactgcttcc agaaggacat ggccatgtcc ttctggaagc agtagagcag 60
cccg 64
<210>548
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>548
cgggctgctc tactgcttca ggaacgacat ggacaaggcc ttgtccatgt cgttcctgaa 60
gcagtagagc agcccg 76
<210>549
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>549
cgggctgctc tactgcttca ggcaggacat ggacaaggcc ttgtccatgt cctgcctgaa 60
gcagtagagc agcccg 76
<210>550
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>550
cgggctgctc tactgcttca ggaagaacat ggacaaggcc ttgtccatgt tcttcctgaa 60
gcagtagagc agcccg 76
<210>551
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>551
cgggctgctc tactgcttca ggaagcagat ggacaaggcc ttgtccatct gcttcctgaa 60
gcagtagagc agcccg 76
<210>552
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>552
caggaaggac atcgacaagg tcgagacatt cctgcgcgcg caggaatgtc tcgaccttgt 60
cgatgtcctt cctg 74
<210>553
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>553
caggaaggac gtggacaagg tcgagacatt cctgcgcgcg caggaatgtc tcgaccttgt 60
ccacgtcctt cctg 74
<210>554
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>554
ggaaggacat gaacaaggtc gagacattcc tgcgcgcgca ggaatgtctc gaccttgttc 60
atgtccttcc 70
<210>555
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>555
ggaaggacat gcagaaggtc gagacattcc tgcgcgcgca ggaatgtctc gaccttctgc 60
atgtccttcc 70
<210>556
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>556
ggacatggac aacgtcgaga cattcctgcg cgcgcaggaa tgtctcgacg ttgtccatgt 60
cc 62
<210>557
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>557
ggacatggac caggtcgaga cattcctgcg cgcgcaggaa tgtctcgacc tggtccatgt 60
cc 62
<210>558
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>558
ggacaaggtc cagacattcc tgcgcatcgt gcagtgccgc ggcactgcac gatgcgcagg 60
aatgtctgga ccttgtcc 78
<210>559
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>559
ggacaaggtc cacacattcc tgcgcatcgt gcagtgccgc ggcactgcac gatgcgcagg 60
aatgtgtgga ccttgtcc 78
<210>560
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>560
ggacaaggtc aacacattcc tgcgcatcgt gcagtgccgc ggcactgcac gatgcgcagg 60
aatgtgttga ccttgtcc 78
<210>561
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>561
ggtcgagaca atcctgcgca tcgtgcagtg ccgcgcggca ctgcacgatg cgcaggattg 60
tctcgacc 68
<210>562
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>562
ggtcgagaca gtgctgcgca tcgtgcagtg ccgcgcggca ctgcacgatg cgcagcactg 60
tctcgacc 68
<210>563
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>563
ggtcgagaca ttcatccgca tcgtgcagtg ccgcgcggca ctgcacgatg cggatgaatg 60
tctcgacc 68
<210>564
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>564
ggtcgagaca ttcgtgcgca tcgtgcagtg ccgcgcggca ctgcacgatg cgcacgaatg 60
tctcgacc 68
<210>565
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>565
ggtcgagaca ttcctgcaca tcgtgcagtg ccgcgcggca ctgcacgatg tgcaggaatg 60
tctcgacc 68
<210>566
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>566
ggtcgagaca ttcctgcaga tcgtgcagtg ccgcgcggca ctgcacgatc tgcaggaatg 60
tctcgacc 68
<210>567
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>567
gcgcatcgtg cagtgccact ctgtggaggg cgccctccac agagtggcac tgcacgatgc 60
gc 62
<210>568
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>568
gcgcatcgtg cagtgccagt ctgtggaggg cgccctccac agactggcac tgcacgatgc 60
gc 62
<210>569
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>569
gcagtgccgc tctgtgcagg gcagctgtgg cgccacagct gccctgcaca gagcggcact 60
gc 62
<210>570
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>570
gcagtgccgc tctgtgcacg gcagctgtgg cgccacagct gccgtgcaca gagcggcact 60
gc 62
<210>571
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>571
gcagtgccgc tctgtgaacg gcagctgtgg cgccacagct gccgttcaca gagcggcact 60
gc 62
<210>572
<211>71
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>572
gccgctctgt ggagggcagc tgtggcatct agagggcccg ccacccgggc agctagatgc 60
cacagctgcc c 71
<210>573
<211>71
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>573
gccgctctgt ggagggcagc tgtggcgtgt agagggcccg ccacccgggc agctacacgc 60
cacagctgcc c 71
<210>574
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>574
ggcagtgcct tcccaaccat tcccgagtcc aggctttttg caaaaagcct ggactcggga 60
atggttggga aggcactgcc 80
<210>575
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>575
ggcagtgcct tcccaaccat tcccgactcc aggctttttg caaaaagcct ggagtcggga 60
atggttggga aggcactgcc 80
<210>576
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>576
ggcagtgcct tcccaaccat tcccaagtcc aggctttttg caaaaagcct ggacttggga 60
atggttggga aggcactgcc 80
<210>577
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>577
ggcagtgcct tcccaaccat tcccaggtcc aggctttttg caaaaagcct ggacctggga 60
atggttggga aggcactgcc 80
<210>578
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>578
ggcagtgcct tcccaaccat tcccaactcc aggctttttg caaaaagcct ggagttggga 60
atggttggga aggcactgcc 80
<210>579
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>579
ggcagtgcct tcccaaccat tccccagtcc aggctttttg caaaaagcct ggactgggga 60
atggttggga aggcactgcc 80
<210>580
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>580
ggcagtgcct tcccaaccat tcccagctcc aggctttttg caaaaagcct ggagctggga 60
atggttggga aggcactgcc 80
<210>581
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>581
ggcagtgcct tcccaaccat tcccacctcc aggctttttg caaaaagcct ggaggtggga 60
atggttggga aggcactgcc 80
<210>582
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>582
cccttatcca gggagtttga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tcaaactccc tggataaggg 80
<210>583
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>583
cccttatcca gggactttga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tcaaagtccc tggataaggg 80
<210>584
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>584
cccttatcca ggaagtttga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tcaaacttcc tggataaggg 80
<210>585
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>585
cccttatcca ggaggtttga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tcaaacctcc tggataaggg 80
<210>586
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>586
cccttatcca ggaactttga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tcaaagttcc tggataaggg 80
<210>587
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>587
cccttatcca ggcagtttga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tcaaactgcc tggataaggg 80
<210>588
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>588
cccttatcca ggagctttga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tcaaagctcc tggataaggg 80
<210>589
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>589
cccttatcca ggacctttga caacgctatg ctccgcgccc gggcgcggag catagcgttg 60
tcaaaggtcc tggataaggg 80
<210>590
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>590
ccaggctttt tgacaacgag atgctccgcg cccgggcgcg gagcatctcg ttgtcaaaaa 60
gcctgg 66
<210>591
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>591
ccaggctttt tgacaacgac atgctccgcg cccgggcgcg gagcatgtcg ttgtcaaaaa 60
gcctgg 66
<210>592
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>592
ccaggctttt tgacaacaag atgctccgcg cccgggcgcg gagcatcttg ttgtcaaaaa 60
gcctgg 66
<210>593
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>593
ccaggctttt tgacaacagg atgctccgcg cccgggcgcg gagcatcctg ttgtcaaaaa 60
gcctgg 66
<210>594
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>594
ccaggctttt tgacaacaac atgctccgcg cccgggcgcg gagcatgttg ttgtcaaaaa 60
gcctgg 66
<210>595
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>595
ccaggctttt tgacaaccag atgctccgcg cccgggcgcg gagcatctgg ttgtcaaaaa 60
gcctgg 66
<210>596
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>596
ccaggctttt tgacaacagc atgctccgcg cccgggcgcg gagcatgctg ttgtcaaaaa 60
gcctgg 66
<210>597
<211>66
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>597
ccaggctttt tgacaacacc atgctccgcg cccgggcgcg gagcatggtg ttgtcaaaaa 60
gcctgg 66
<210>598
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>598
caacgctatg gagcgcgccc atcgtctgca ccagctggcg ccagctggtg cagacgatgg 60
gcgcgctcca tagcgttg 78
<210>599
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>599
caacgctatg gaccgcgccc atcgtctgca ccagctggcg ccagctggtg cagacgatgg 60
gcgcggtcca tagcgttg 78
<210>600
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>600
caacgctatg aagcgcgccc atcgtctgca ccagctggcg ccagctggtg cagacgatgg 60
gcgcgcttca tagcgttg 78
<210>601
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>601
caacgctatg aggcgcgccc atcgtctgca ccagctggcg ccagctggtg cagacgatgg 60
gcgcgcctca tagcgttg 78
<210>602
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>602
caacgctatg aaccgcgccc atcgtctgca ccagctggcg ccagctggtg cagacgatgg 60
gcgcggttca tagcgttg 78
<210>603
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>603
caacgctatg cagcgcgccc atcgtctgca ccagctggcg ccagctggtg cagacgatgg 60
gcgcgctgca tagcgttg 78
<210>604
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>604
caacgctatg agccgcgccc atcgtctgca ccagctggcg ccagctggtg cagacgatgg 60
gcgcggctca tagcgttg 78
<210>605
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>605
caacgctatg acccgcgccc atcgtctgca ccagctggcg ccagctggtg cagacgatgg 60
gcgcgggtca tagcgttg 78
<210>606
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>606
cgctatgctc cgcgagcatc gtctgcacca gctggccggc cagctggtgc agacgatgct 60
cgcggagcat agcg 74
<210>607
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>607
cgctatgctc cgcgaccatc gtctgcacca gctggccggc cagctggtgc agacgatggt 60
cgcggagcat agcg 74
<210>608
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>608
cgctatgctc cgcaagcatc gtctgcacca gctggccggc cagctggtgc agacgatgct 60
tgcggagcat agcg 74
<210>609
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>609
cgctatgctc cgcaggcatc gtctgcacca gctggccggc cagctggtgc agacgatgcc 60
tgcggagcat agcg 74
<210>610
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>610
cgctatgctc cgcaaccatc gtctgcacca gctggccggc cagctggtgc agacgatggt 60
tgcggagcat agcg 74
<210>611
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>611
cgctatgctc cgccagcatc gtctgcacca gctggccggc cagctggtgc agacgatgct 60
ggcggagcat agcg 74
<210>612
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>612
cgctatgctc cgcagccatc gtctgcacca gctggccggc cagctggtgc agacgatggc 60
tgcggagcat agcg 74
<210>613
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>613
cgctatgctc cgcacccatc gtctgcacca gctggccggc cagctggtgc agacgatggg 60
tgcggagcat agcg 74
<210>614
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>614
gctccgcgcc catcgtgagc accagctggc cggccagctg gtgctcacga tgggcgcgga 60
gc 62
<210>615
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>615
gctccgcgcc catcgtgacc accagctggc cggccagctg gtggtcacga tgggcgcgga 60
gc 62
<210>616
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>616
gctccgcgcc catcgtaagc accagctggc cggccagctg gtgcttacga tgggcgcgga 60
gc 62
<210>617
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>617
gctccgcgcc catcgtaggc accagctggc cggccagctg gtgcctacga tgggcgcgga 60
gc 62
<210>618
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>618
gctccgcgcc catcgtaacc accagctggc cggccagctg gtggttacga tgggcgcgga 60
gc 62
<210>619
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>619
gctccgcgcc catcgtcagc accagctggc cggccagctg gtgctgacga tgggcgcgga 60
gc 62
<210>620
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>620
gctccgcgcc catcgtagcc accagctggc cggccagctg gtggctacga tgggcgcgga 60
gc 62
<210>621
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>621
gctccgcgcc catcgtaccc accagctggc cggccagctg gtgggtacga tgggcgcgga 60
gc 62
<210>622
<211>74
<212>DN
<213>Artificial Sequence
<220>
<223>Primer
<400>622
ccgcgcccat cgtctgcacc aggaggcctt tgacaccggt gtcaaaggcc tcctggtgca 60
gacgatgggc gcgg 74
<210>623
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>623
ccgcgcccat cgtctgcacc aggacgcctt tgacaccggt gtcaaaggcg tcctggtgca 60
gacgatgggc gcgg 74
<210>624
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>624
gctccgcgcc catcgtctgc accagaaggc ctttgacacc ggtgtcaaag gccttctggt 60
gcagacgatg ggcgcggagc 80
<210>625
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>625
ccgcgcccat cgtctgcacc agagggcctt tgacaccggt gtcaaaggcc ctctggtgca 60
gacgatgggc gcgg 74
<210>626
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>626
gctccgcgcc catcgtctgc accagaacgc ctttgacacc ggtgtcaaag gcgttctggt 60
gcagacgatg ggcgcggagc 80
<210>627
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>627
ccgcgcccat cgtctgcacc agcaggcctt tgacaccggt gtcaaaggcc tgctggtgca 60
gacgatgggc gcgg 74
<210>628
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>628
ccgcgcccat cgtctgcacc agagcgcctt tgacaccggt gtcaaaggcg ctctggtgca 60
gacgatgggc gcgg 74
<210>629
<211>74
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>629
ccgcgcccat cgtctgcacc agaccgcctt tgacaccggt gtcaaaggcg gtctggtgca 60
gacgatgggc gcgg 74
<210>630
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>630
cgcgcccatc gtctgcacca gctggagttt gacacctacc ggtaggtgtc aaactccagc 60
tggtgcagac gatgggcgcg 80
<210>631
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>631
cgcgcccatc gtctgcacca gctggacttt gacacctacc ggtaggtgtc aaagtccagc 60
tggtgcagac gatgggcgcg 80
<210>632
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>632
cgcgcccatc gtctgcacca gctgaagttt gacacctacc ggtaggtgtc aaacttcagc 60
tggtgcagac gatgggcgcg 80
<210>633
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>633
cgcgcccatc gtctgcacca gctgaggttt gacacctacc ggtaggtgtc aaacctcagc 60
tggtgcagac gatgggcgcg 80
<210>634
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>634
cgcgcccatc gtctgcacca gctgaacttt gacacctacc ggtaggtgtc aaagttcagc 60
tggtgcagac gatgggcgcg 80
<210>635
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>635
cgcgcccatc gtctgcacca gctgcagttt gacacctacc ggtaggtgtc aaactgcagc 60
tggtgcagac gatgggcgcg 80
<210>636
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>636
cgcgcccatc gtctgcacca gctgagcttt gacacctacc ggtaggtgtc aaagctcagc 60
tggtgcagac gatgggcgcg 80
<210>637
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>637
cgcgcccatc gtctgcacca gctgaccttt gacacctacc ggtaggtgtc aaaggtcagc 60
tggtgcagac gatgggcgcg 80
<210>638
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>638
gcctggtgta cggcgagtct gacagcaacg cgttgctgtc agactcgccg tacaccaggc 60
<210>639
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>639
gcctggtgta cggcgactct gacagcaacg cgttgctgtc agagtcgccg tacaccaggc 60
<210>640
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>640
cgccaacagc ctggtgtacg gcaagtctga cagcaacgcg ttgctgtcag acttgccgta 60
caccaggctg ttggcg 76
<210>641
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>641
gcctggtgta cggcaggtct gacagcaacg cgttgctgtc agacctgccg tacaccaggc 60
<210>642
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>642
cgccaacagc ctggtgtacg gcaactctga cagcaacgcg ttgctgtcag agttgccgta 60
caccaggctg ttggcg 76
<210>643
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>643
gcctggtgta cggccagtct gacagcaacg cgttgctgtc agactggccg tacaccaggc 60
<210>644
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>644
gcctggtgta cggcagctct gacagcaacg cgttgctgtc agagctgccg tacaccaggc 60
<210>645
<211>60
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>645
gcctggtgta cggcacctct gacagcaacg cgttgctgtc agaggtgccg tacaccaggc 60
<210>646
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>646
cggcgcctct gacagcaacg agtatgacct ccggaggtca tactcgttgc tgtcagaggc 60
gccg 64
<210>647
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>647
cggcgcctct gacagcaacg actatgacct ccggaggtca tagtcgttgc tgtcagaggc 60
gccg 64
<210>648
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>648
cggcgcctct gacagcaaca agtatgacct ccggaggtca tacttgttgc tgtcagaggc 60
gccg 64
<210>649
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>649
cggcgcctct gacagcaaca ggtatgacct ccggaggtca tacctgttgc tgtcagaggc 60
gccg 64
<210>650
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>650
cggcgcctct gacagcaaca actatgacct ccggaggtca tagttgttgc tgtcagaggc 60
gccg 64
<210>651
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>651
cggcgcctct gacagcaacc agtatgacct ccggaggtca tactggttgc tgtcagaggc 60
gccg 64
<210>652
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>652
cggcgcctct gacagcaaca gctatgacct ccggaggtca tagctgttgc tgtcagaggc 60
gccg 64
<210>653
<211>64
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>653
cggcgcctct gacagcaaca cctatgacct ccggaggtca taggtgttgc tgtcagaggc 60
gccg 64
<210>654
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>654
gcgcctctga cagcaacgtc tatgacgagc taaaggaccg gtcctttagc tcgtcataga 60
cgttgctgtc agaggcgc 78
<210>655
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>655
gcgcctctga cagcaacgtc tatgacgacc taaaggaccg gtcctttagg tcgtcataga 60
cgttgctgtc agaggcgc 78
<210>656
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>656
gcgcctctga cagcaacgtc tatgacaagc taaaggaccg gtcctttagc ttgtcataga 60
cgttgctgtc agaggcgc 78
<210>657
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>657
gcgcctctga cagcaacgtc tatgacaggc taaaggaccg gtcctttagc ctgtcataga 60
cgttgctgtc agaggcgc 78
<210>658
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>658
gcgcctctga cagcaacgtc tatgacaacc taaaggaccg gtcctttagg ttgtcataga 60
cgttgctgtc agaggcgc 78
<210>659
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>659
gcgcctctga cagcaacgtc tatgaccagc taaaggaccg gtcctttagc tggtcataga 60
cgttgctgtc agaggcgc 78
<210>660
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>660
gcgcctctga cagcaacgtc tatgacagcc taaaggaccg gtcctttagg ctgtcataga 60
cgttgctgtc agaggcgc 78
<210>661
<211>78
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>661
gcgcctctga cagcaacgtc tatgacaccc taaaggaccg gtcctttagg gtgtcataga 60
cgttgctgtc agaggcgc 78
<210>662
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>662
cgtctatgac ctcgagaagg acctagagga aggcgccttc ctctaggtcc ttctcgaggt 60
catagacg 68
<210>663
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>663
cgtctatgac ctcgacaagg acctagagga aggcgccttc ctctaggtcc ttgtcgaggt 60
catagacg 68
<210>664
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>664
cgtctatgac ctcaagaagg acctagagga aggcgccttc ctctaggtcc ttcttgaggt 60
catagacg 68
<210>665
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>665
cgtctatgac ctcaggaagg acctagagga aggcgccttc ctctaggtcc ttcctgaggt 60
catagacg 68
<210>666
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>666
cgtctatgac ctcaacaagg acctagagga aggcgccttc ctctaggtcc ttgttgaggt 60
catagacg 68
<210>667
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>667
cgtctatgac ctccagaagg acctagagga aggcgccttc ctctaggtcc ttctggaggt 60
catagacg 68
<210>668
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>668
cgtctatgac ctcagcaagg acctagagga aggcgccttc ctctaggtcc ttgctgaggt 60
catagacg 68
<210>669
<211>68
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>669
cgtctatgac ctcaccaagg acctagagga aggcgccttc ctctaggtcc ttggtgaggt 60
catagacg 68
<210>670
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>670
gacctcctaa aggacgagga ggaaggcatc cggatgcctt cctcctcgtc ctttaggagg 60
tc 62
<210>671
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>671
gacctcctaa aggacgacga ggaaggcatc cggatgcctt cctcgtcgtc ctttaggagg 60
tc 62
<210>672
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>672
gacctcctaa aggacaagga ggaaggcatc cggatgcctt cctccttgtc ctttaggagg 60
tc 62
<210>673
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>673
gacctcctaa aggacaggga ggaaggcatc cggatgcctt cctccctgtc ctttaggagg 60
tc 62
<210>674
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>674
gacctcctaa aggacaacga ggaaggcatc cggatgcctt cctcgttgtc ctttaggagg 60
tc 62
<210>675
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>675
gacctcctaa aggaccagga ggaaggcatc cggatgcctt cctcctggtc ctttaggagg 60
tc 62
<210>676
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>676
gacctcctaa aggacagcga ggaaggcatc cggatgcctt cctcgctgtc ctttaggagg 60
tc 62
<210>677
<211>62
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>677
gacctcctaa aggacaccga ggaaggcatc cggatgcctt cctcggtgtc ctttaggagg 60
tc 62
<210>678
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>678
cctagaggaa ggcgagcaaa cgctgatggg gaggctggcc agcctcccca tcagcgtttg 60
ctcgccttcc tctagg 76
<210>679
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>679
cctagaggaa ggcgaccaaa cgctgatggg gaggctggcc agcctcccca tcagcgtttg 60
gtcgccttcc tctagg 76
<210>680
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>680
cctagaggaa ggcaagcaaa cgctgatggg gaggctggcc agcctcccca tcagcgtttg 60
cttgccttcc tctagg 76
<210>681
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>681
cctagaggaa ggcaggcaaa cgctgatggg gaggctggcc agcctcccca tcagcgtttg 60
cctgccttcc tctagg 76
<210>682
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>682
cctagaggaa ggcaaccaaa cgctgatggg gaggctggcc agcctcccca tcagcgtttg 60
gttgccttcc tctagg 76
<210>683
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>683
cctagaggaa ggccagcaaa cgctgatggg gaggctggcc agcctcccca tcagcgtttg 60
ctggccttcc tctagg 76
<210>684
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>684
cctagaggaa ggcagccaaa cgctgatggg gaggctggcc agcctcccca tcagcgtttg 60
gctgccttcc tctagg 76
<210>685
<211>76
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>685
cctagaggaa ggcacccaaa cgctgatggg gaggctggcc agcctcccca tcagcgtttg 60
ggtgccttcc tctagg 76
<210>686
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>686
gcatccaaac ggagatgggg aggctggaag atggcagccc gggctgccat cttccagcct 60
ccccatctcc gtttggatgc 80
<210>687
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>687
gcatccaaac ggacatgggg aggctggaag atggcagccc gggctgccat cttccagcct 60
ccccatgtcc gtttggatgc 80
<210>688
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>688
gcatccaaac gaagatgggg aggctggaag atggcagccc gggctgccat cttccagcct 60
ccccatcttc gtttggatgc 80
<210>689
<211>79
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>689
gcatccaaac gaggatgggg aggctggaag atggcagccc ggctgccatc ttccagcctc 60
cccatcctcg tttggatgc 79
<210>690
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>690
gcatccaaac gaacatgggg aggctggaag atggcagccc gggctgccat cttccagcct 60
ccccatgttc gtttggatgc 80
<210>691
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>691
gcatccaaac gcagatgggg aggctggaag atggcagccc gggctgccat cttccagcct 60
ccccatctgc gtttggatgc 80
<210>692
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>692
gcatccaaac gagcatgggg aggctggaag atggcagccc gggctgccat cttccagcct 60
ccccatgctc gtttggatgc 80
<210>693
<211>80
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>693
gcatccaaac gaccatgggg aggctggaag atggcagccc gggctgccat cttccagcct 60
ccccatggtc gtttggatgc 80
<210>694
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>694
cgctgatggg gagggaggaa gatggcagcc cccggccggg ggctgccatc ttcctccctc 60
cccatcagcg 70
<210>695
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>695
cgctgatggg gagggacgaa gatggcagcc cccggccggg ggctgccatc ttcgtccctc 60
cccatcagcg 70
<210>696
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>696
cgctgatggg gaggaaggaa gatggcagcc cccggccggg ggctgccatc ttccttcctc 60
cccatcagcg 70
<210>697
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>697
cgctgatggg gaggagggaa gatggcagcc cccggccggg ggctgccatc ttccctcctc 60
cccatcagcg 70
<210>698
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>698
cgctgatggg gaggaacgaa gatggcagcc cccggccggg ggctgccatc ttcgttcctc 60
cccatcagcg 70
<210>699
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>699
cgctgatggg gaggcaggaa gatggcagcc cccggccggg ggctgccatc ttcctgcctc 60
cccatcagcg 70
<210>700
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>700
cgctgatggg gaggagcgaa gatggcagcc cccggccggg ggctgccatc ttcgctcctc 60
cccatcagcg 70
<210>701
<211>70
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>701
cgctgatggg gaggaccgaa gatggcagcc cccggccggg ggctgccatc ttcggtcctc 60
cccatcagcg 70
<210>702
<211>32
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>702
atggctacag gctcccggac gtccctgcct cc 32
<210>703
<211>37
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>703
ctagaagcca caagctgccc ctccacagag cggcact 37
<210>704
<211>30
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>704
aaaaagctta tggctacagg ctcccggacg 30
<210>705
<211>32
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>705
gcccttctag aagccacagc tgccctccac ag 32
<210>706
<211>38
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>706
gggaattcca tatgttcccg accattccgt tatccagg 38
<210>707
<211>39
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>707
cgggatccct taagaagcca cagctgccct ccacagagc 39
<210>708
<211>31
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>708
ccgaccattc cgttacccgt ctttttgaca a 31
<210>709
<211>32
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>709
gagcatagcg ttgtcaaaaa gacgggataa cg 32
<210>710
<211>36
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>710
gacaacgcta tgctccgtgc ccatcgtctg caccag 36
<210>711
<211>36
<212>DNA
<213>Artificial Sequence
<220>
<223>Primer
<400>711
ctggtgcaga cgatgggcac ggagcatagc gttgtc 36
<210>712
<211>217
<212>PRT
<213>Homo sapiens
<300>
<308>GenBank NP_002050.1
<309>2004-10-26
<400>712
Met Ala Ala Gly Ser Arg Thr Ser Leu Leu Leu Ala Phe Gly Leu Leu
1 5 10 15
Cys Leu Ser Trp Leu Gln Glu Gly Ser Ala Phe Pro Thr Ile Pro Leu
20 25 30
Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala Arg Arg Leu Tyr Gln
35 40 45
Leu Ala Tyr Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Leu Lys
50 55 60
Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe
65 70 75 80
Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Val Lys Thr Gln Gln Lys
85 90 95
Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp
100 105 110
Leu Glu Pro Val Gln Leu Leu Arg Ser Val Phe Ala Asn Ser Leu Val
115 120 125
Tyr Gly Ala Ser Asp Ser Asn Val Tyr Arg His Leu Lys Asp Leu Glu
130 135 140
Glu Gly Ile Gln Thr Leu Met Trp Arg Leu Glu Asp Gly Ser Pro Arg
145 150 155 160
Thr Gly Gln Ile Phe Asn Gln Ser Tyr Ser Lys Phe Asp Thr Lys Ser
165 170 175
His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe
180 185 190
Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys
195 200 205
Arg Ser Val Glu Gly Ser Cys Gly Phe
210 215
<210>713
<211>217
<212>PRT
<213>Homo sapiens
<300>
<308>GenBank NP_000506.2
<309>2004-10-26
<400>713
Met Ala Thr Gly Ser Arg Thr Ser Leu Leu Leu Ala Phe Gly Leu Leu
1 5 10 15
Cys Leu Pro Trp Leu Gln Glu Gly Ser Ala Phe Pro Thr Ile Pro Leu
20 25 30
Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln
35 40 45
Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys
50 55 60
Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe
65 70 75 80
Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys
85 90 95
Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp
100 105 110
Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val
115 120 125
Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu
130 135 140
Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg
145 150 155 160
Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser
165 170 175
His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe
180 185 190
Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys
195 200 205
Arg Ser Val Glu Gly Ser Cys Gly Phe
210 215
<210>714
<211>217
<212>PRT
<213>Bos taurus
<300>
<308>GenBank P01246
<309>1986-07-21
<400>714
Met Met Ala Ala Gly Pro Arg Thr Ser Leu Leu Leu Ala Phe Ala Leu
1 5 10 15
Leu Cys Leu Pro Trp Thr Gln Val Val Gly Ala Phe Pro Ala Met Ser
20 25 30
Leu Ser Gly Leu Phe Ala Asn Ala Val Leu Arg Ala Gln His Leu His
35 40 45
Gln Leu Ala Ala Asp Thr Phe Lys Glu Phe Glu Arg Thr Tyr Ile Pro
50 55 60
Glu Gly Gln Arg Tyr Ser Ile Gln Asn Thr Gln Val Ala Phe Cys Phe
65 70 75 80
Ser Glu Thr Ile Pro Ala Pro Thr Gly Lys Asn Glu Ala Gln Gln Lys
85 90 95
Ser Asp Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp
100 105 110
Leu Gly Pro Leu Gln Phe Leu Ser Arg Val Phe Thr Asn Ser Leu Val
115 120 125
Phe Gly Thr Ser Asp Arg Val Tyr Glu Lys Leu Lys Asp Leu Glu Glu
130 135 140
Gly Ile Leu Ala Leu Met Arg Glu Leu Glu Asp Gly Thr Pro Arg Ala
145 150 155 160
Gly Gln Ile Leu Lys Gln Thr Tyr Asp Lys Phe Asp Thr Asn Met Arg
165 170 175
Ser Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Ser Cys Phe Arg
180 185 190
Lys Asp Leu His Lys Thr Glu Thr Tyr Leu Arg Val Met Lys Cys Arg
195 200 205
Arg Phe Gly Glu Ala Ser Cys Ala Phe
210 215
<210>715
<211>157
<212>PRT
<213>Macaca mulatta
<300>
<308>GenBank P33093
<309>1993-10-01
<400>715
Met Ala Ala Gly Ser Arg Thr Ser Leu Leu Leu Ala Phe Ala Leu Leu
1 5 10 15
Cys Leu Pro Trp Leu Gln Glu Gly Ser Ala Phe Pro Thr Ile Pro Leu
20 25 30
Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln
35 40 45
Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys
50 55 60
Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe
65 70 75 80
Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys
85 90 95
Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp
100 105 110
Leu Glu Pro Val Gln Phe Leu Arg Ala Leu Leu Lys Asn Tyr Gly Leu
115 120 125
Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Ile Glu Thr Phe Leu Arg
130 135 140
Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe
145 150 155
<210>716
<211>3821
<212>DNA
<213>Artificial Sequence
<220>
<223>pNAUT hGH vector
<400>716
gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctgcta 60
ggggttcctt gtagttaatg attaacccgc catgctactt atctactatg ccaagtacgc 120
cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 180
tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga 240
tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa 300
gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc 360
caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg 420
aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg 480
aaattaatac gactcactat agggagaccc aagcttatgg ctacaggctc ccggacgtcc 540
ctgctcctgg cttttggcct gctctgcctg ccctggcttc aagagggcag tgccttccca 600
accattccct tatccaggct ttttgacaac gctatgctcc gcgcccatcg tctgcaccag 660
ctggcctttg acacctacca ggagtttgaa gaagcctata tcccaaagga acagaagtat 720
tcattcctgc agaaccccca gacctccctc tgtttctcag agtctattcc gacaccctcc 780
aacagggagg aaacacaaca gaaatccaac ctagagctgc tccgcatctc cctgctgctc 840
atccagtcgt ggctggagcc cgtgcagttc ctcaggagtg tcttcgccaa cagcctggtg 900
tacggcgcct ctgacagcaa cgtctatgac ctcctaaagg acctagagga aggcatccaa 960
acgctgatgg ggaggctgga agatggcagc ccccggactg ggcagatctt caagcagacc 1020
tacagcaagt tcgacacaaa ctcacacaac gatgacgcac tactcaagaa ctacgggctg 1080
ctctactgct tcaggaagga catggacaag gtcgagacat tcctgcgcat cgtgcagtgc 1140
cgctctgtgg agggcagctg tggcttctag agggccctat tctatagtgt cacctaaatg 1200
ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 1260
cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 1320
atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 1380
ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 1440
gctctatggc ttctgaggcg gaaagaacca ggtagataag tagcatggcg ggttaatcat 1500
taactacaca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg 1560
cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt 1620
atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc 1680
cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca 1740
tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg 1800
tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat 1860
gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga 1920
acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 1980
ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 2040
gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 2100
ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 2160
gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 2220
agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 2280
caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 2340
gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 2400
agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 2460
gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 2520
aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 2580
ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 2640
tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 2700
tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 2760
gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 2820
atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 2880
ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 2940
aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 3000
ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 3060
ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 3120
tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 3180
cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct 3240
gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 3300
gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 3360
tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 3420
ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 3480
gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 3540
ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 3600
tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 3660
ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 3720
gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 3780
acgaccgagc gcagcgagtc agtgagcgag gaagcggaag a 3821
<210>717
<211>4611
<212>DNA
<213>Artificial Sequence
<220>
<223>pTOPO hGH vector
<400>717
cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 60
cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 120
acatgtgagc aaaaggccag caaaagccca ggaaccgtaa aaaggccgcg ttgctggcgt 180
ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 240
ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 300
gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 360
gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 420
ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 480
actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 540
gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 600
ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 660
ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 720
gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 780
tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 840
tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 900
aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 960
aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 1020
tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 1080
gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1140
agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1200
aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1260
gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1320
caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1380
cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1440
ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1500
ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1560
gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1620
cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1680
gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1740
caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1800
tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1860
acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1920
aagtgccacc tgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat 1980
caggaaattg taagcgttaa taattcagaa gaactcgtca agaaggcgat agaaggcgat 2040
gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc 2100
gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac 2160
acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg 2220
caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc tcgccttgag 2280
cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc 2340
gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc 2400
gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga 2460
tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa 2520
tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc 2580
cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga 2640
caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc 2700
atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc 2760
ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt 2820
ctcttgatca gagcttgatc ccctgcgcca tcagatcctt ggcggcgaga aagccatcca 2880
gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccggttc 2940
gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac 3000
ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat 3060
ccggggtcag caccgtttct gcggactggc tttctacgtg aaaaggatct aggtgaagat 3120
cctttttgat aatctcatgc ctgacattta tattccccag aacatcaggt taatggcgtt 3180
tttgatgtca ttttcgcggt ggctgagatc agccacttct tccccgataa cggagaccgg 3240
cacactggcc atatcggtgg tcatcatgcg ccagctttca tccccgatat gcaccaccgg 3300
gtaaagttca cgggagactt tatctgacag cagacgtgca ctggccaggg ggatcaccat 3360
ccgtcgcccc ggcgtgtcaa taatatcact ctgtacatcc acaaacagac gataacggct 3420
ctctctttta taggtgtaaa ccttaaactg ccgtacgtat aggctgcgca actgttggga 3480
agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 3540
aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 3600
cagtgaattg taatacgact cactataggg cgaattgaat ttagcggccg cgaattcgcc 3660
cttatggcta caggctcccg gacgtccctg ctcctggctt ttggcctgct ctgcctgccc 3720
tggcttcaag agggcagtgc cttcccaacc attcccttat ccaggctttt tgacaacgct 3780
atgctccgcg cccatcgtct gcaccagctg gcctttgaca cctaccagga gtttgaagaa 3840
gcctatatcc caaaggaaca gaagtattca ttcctgcaga acccccagac ctccctctgt 3900
ttctcagagt ctattccgac accctccaac agggaggaaa cacaacagaa atccaaccta 3960
gagctgctcc gcatctccct gctgctcatc cagtcgtggc tggagcccgt gcagttcctc 4020
aggagtgtct tcgccaacag cctggtgtac ggcgcctctg acagcaacgt ctatgacctc 4080
ctaaaggacc tagaggaagg catccaaacg ctgatgggga ggctggaaga tggcagcccc 4140
cggactgggc agatcttcaa gcagacctac agcaagttcg acacaaactc acacaacgat 4200
gacgcactac tcaagaacta cgggctgctc tactgcttca ggaaggacat ggacaaggtc 4260
gagacattcc tgcgcatcgt gcagtgccgc tctgtggagg gcagctgtgg cttctagaag 4320
ggcgaattcg tttaaacctg caggactagt ccctttagtg agggttaatt ctgagcttgg 4380
cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 4440
acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 4500
cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 4560
attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc t 4611
<210>718
<211>5236
<212>DNA
<213>Artificial Sequence
<220>
<223>pET-24 vector
<400>718
atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 60
ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 120
tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt 180
cgacggagct cgaattcgga tcctagaggg gaattgttat ccgctcacaa ttcccctata 240
gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct ctacgccgga cgcatcgtgg 300
ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg 360
gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg 420
caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg 480
cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata 540
agggagagcg tcgagatccc ggacaccatc gaatggcgca aaacctttcg cggtatggca 600
tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac 660
gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc 720
agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac 780
attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc 840
acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc 900
gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt 960
aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg 1020
ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt 1080
cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg 1140
cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc 1200
ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc 1260
aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa 1320
caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat 1380
cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat 1440
atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc 1500
accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc 1560
tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa 1620
accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg 1680
cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt 1740
aagttagctc actcattagg caccgggatc tcgaccgatg cccttgagag ccttcaaccc 1800
agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt 1860
ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga 1920
ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca 1980
cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc 2040
cattatcgcc ggcatggcgg ccccacgggt gcgcatgatc gtgctcctgt cgttgaggac 2100
ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg 2160
aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt 2220
cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt 2280
ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac 2340
gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag 2400
ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt 2460
gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac 2520
ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag 2580
ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat 2640
ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg 2700
tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 2760
agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 2820
gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg 2880
gcatcagagc agattgtact gagagtgcac catatatgcg gtgtgaaata ccgcacagat 2940
gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc 3000
gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 3060
ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 3120
ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 3180
atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 3240
aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 3300
gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 3360
ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 3420
ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 3480
acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 3540
gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat 3600
ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 3660
ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 3720
gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 3780
ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc tgcttacata 3840
aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg ctctaggccg 3900
cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc 3960
gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt 4020
ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac 4080
tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat 4140
gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat 4200
cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg 4260
attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa 4320
tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg 4380
cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc 4440
gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt 4500
tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg 4560
aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt 4620
gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaagaa 4680
ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 4740
gcgcacattt ccccgaaaag tgccacctga aattgtaaac gttaatattt tgttaaaatt 4800
cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 4860
cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa 4920
gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 4980
cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa 5040
agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc 5100
gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag 5160
tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg 5220
cgcgtcccat tcgcca 5236
<210>719
<211>10574
<212>DNA
<213>Artificial Sequence
<220>
<223>pET24Naut-hGH Plasmid
<400>719
ctagtcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata gatcagcccc 60
tttacacgcg ccttggggat aaacaaataa aaagatttat cattcaaata tgtatccgct 120
catgaattaa ttcttagaaa aactcatcga gtaagtttat acataggcga gtacttaatt 180
aagaatcttt ttgagtagct gcatcaaatg aaactgcaat ttattcatat caggattatc 240
aataccatat cgtagtttac tttgacgtta aataagtata gtcctaatag ttatggtata 300
ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt aaaacttttt 360
cggcaaagac attacttcct cttttgagtg gctccgtcaa ccataggatg gcaagatcct 420
ggtatcggtc tgcgattccg actcgtccaa ggtatcctac cgttctagga ccatagccag 480
acgctaaggc tgagcaggtt catcaataca acctattaat ttcccctcgt caaaaataag 540
gttatcaagt gtagttatgt tggataatta aaggggagca gtttttattc caatagttca 600
gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt ctctttagtg 660
gtactcactg ctgacttagg ccactcttac cgttttcaaa atgcatttct ttccagactt 720
gttcaacagg ccagccatta cgctcgtcat tacgtaaaga aaggtctgaa caagttgtcc 780
ggtcggtaat gcgagcagta caaaatcact cgcatcaacc aaaccgttat tcattcgtga 840
ttgcgcctga gttttagtga gcgtagttgg tttggcaata agtaagcact aacgcggact 900
gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgctctgctt 960
tatgcgctag cgacaatttt cctgttaatg tttgtcctta cgaatgcaac cggcgcagga 1020
acactgccag cgcatcaaca atattttcac gcttacgttg gccgcgtcct tgtgacggtc 1080
gcgtagttgt tataaaagtg ctgaatcagg atattcttct aatacctgga atgctgtttt 1140
cccggggatc gacttagtcc tataagaaga ttatggacct tacgacaaaa gggcccctag 1200
gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat cgtcaccact 1260
cattggtacg tagtagtcct catgcctatt ttacgaacta ggtcggaaga ggcataaatt 1320
ccgtcagcca gtttagtctg accatctcat ccagccttct ccgtatttaa ggcagtcggt 1380
caaatcagac tggtagagta ctgtaacatc attggcaacg ctacctttgc catgtttcag 1440
aaacaactct gacattgtag taaccgttgc gatggaaacg gtacaaagtc tttgttgaga 1500
ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc ccgcgtagcc 1560
cgaagggtat gttagctatc taacagcgtg gactaacggg gacattatcg cgagcccatt 1620
tatacccata taaatcagca tccatgttgg ctgtaatagc gctcgggtaa atatgggtat 1680
atttagtcgt aggtacaacc aatttaatcg cggcctagag caagacgttt cccgttgaat 1740
atggctcata ttaaattagc gccggatctc gttctgcaaa gggcaactta taccgagtat 1800
acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgtggggaac 1860
ataatgacaa atacattcgt ctgtcaaaat aacaagtact ccaaaatccc ttaacgtgag 1920
ttttcgttcc actgagcgtc agaccccgta ggttttaggg aattgcactc aaaagcaagg 1980
tgactcgcag tctggggcat gaaaagatca aaggatcttc ttgagatcct ttttttctgc 2040
gcgtaatctg cttttctagt ttcctagaag aactctagga aaaaaagacg cgcattagac 2100
ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg gacgaacgtt 2160
tgtttttttg gtggcgatgg tcgccaccaa acaaacggcc atcaagagct accaactctt 2220
tttccgaagg taactggctt cagcagagcg tagttctcga tggttgagaa aaaggcttcc 2280
attgaccgaa gtcgtctcgc cagataccaa atactgtcct tctagtgtag ccgtagttag 2340
gccaccactt gtctatggtt tatgacagga agatcacatc ggcatcaatc cggtggtgaa 2400
caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac gttcttgaga 2460
catcgtggcg gatgtatgga gcgagacgat taggacaatg cagtggctgc tgccagtggc 2520
gataagtcgt gtcttaccgg gttggactca gtcaccgacg acggtcaccg ctattcagca 2580
cagaatggcc caacctgagt agacgatagt taccggataa ggcgcagcgg tcgggctgaa 2640
cggggggttc tctgctatca atggcctatt ccgcgtcgcc agcccgactt gccccccaag 2700
gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc cacgtgtgtc 2760
gggtcgaacc tcgcttgctg gatgtggctt gactctatgg tacagcgtga gctatgagaa 2820
agcgccacgc ttcccgaagg gagaaaggcg atgtcgcact cgatactctt tcgcggtgcg 2880
aagggcttcc ctctttccgc gacaggtatc cggtaagcgg cagggtcgga acaggagagc 2940
gcacgaggga ctgtccatag gccattcgcc gtcccagcct tgtcctctcg cgtgctccct 3000
gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc cgaaggtccc 3060
cctttgcgga ccatagaaat atcaggacag cccaaagcgg acctctgact tgagcgtcga 3120
tttttgtgat gctcgtcagg ggggcggagc tggagactga actcgcagct aaaaacacta 3180
cgagcagtcc ccccgcctcg ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 3240
tggccttttg gatacctttt tgcggtcgtt gcgccggaaa aatgccaagg accggaaaac 3300
ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg gaccggaaaa 3360
cgagtgtaca agaaaggacg caatagggga ctaagacacc ataaccgtat taccgccttt 3420
gagtgagctg ataccgctcg ccgcagccga tattggcata atggcggaaa ctcactcgac 3480
tatggcgagc ggcgtcggct acgaccgagc gcagcgagtc agtgagcgag gaagcggaag 3540
agcgcctgat tgctggctcg cgtcgctcag tcactcgctc cttcgccttc tcgcggacta 3600
gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cgccataaaa 3660
gaggaatgcg tagacacgcc ataaagtgtg gcgttaccac cactctcagt acaatctgct 3720
ctgatgccgc atagttaagc cagtatacac gtgagagtca tgttagacga gactacggcg 3780
tatcaattcg gtcatatgtg tccgctatcg ctacgtgact gggtcatggc tgcgccccga 3840
cacccgccaa aggcgatagc gatgcactga cccagtaccg acgcggggct gtgggcggtt 3900
cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac gtgggcgact 3960
gcgcgggact gcccgaacag acgagggccg taggcgaatg agacaagctg tgaccgtctc 4020
cgggagctgc atgtgtcaga ggttttcacc tctgttcgac actggcagag gccctcgacg 4080
tacacagtct ccaaaagtgg gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca 4140
tcagcgtggt cagtagtggc tttgcgcgct ccgtcgacgc catttcgagt agtcgcacca 4200
cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg gcacttcgct 4260
aagtgtctac agacggacaa gtaggcgcag gtcgagcaac agtttctcca gaagcgttaa 4320
tgtctggctt ctgataaagc gggccatgtt tcaaagaggt cttcgcaatt acagaccgaa 4380
gactatttcg cccggtacaa aagggcggtt ttttcctgtt tggtcactga tgcctccgtg 4440
taagggggat ttcccgccaa aaaaggacaa accagtgact acggaggcac attcccccta 4500
ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg aagacaagta 4560
cccccattac tatggctact ttgctctctc ctacgagtgc atacgggtta ctgatgatga 4620
acatgcccgg ttactggaac gttgtgaggg tatgcccaat gactactact tgtacgggcc 4680
aatgaccttg caacactccc taaacaactg gcggtatgga tgcggcggga ccagagaaaa 4740
atcactcagg atttgttgac cgccatacct acgccgccct ggtctctttt tagtgagtcc 4800
gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagttacggt 4860
cgcgaagcaa ttatgtctac atccacaagg tgtcccatcg cagcagcatc ctgcgatgca 4920
gatccggaac ataatggtgc agggcgctga gtcgtcgtag gacgctacgt ctaggccttg 4980
tattaccacg tcccgcgact cttccgcgtt tccagacttt acgaaacacg gaaaccgaag 5040
accattcatg gaaggcgcaa aggtctgaaa tgctttgtgc ctttggcttc tggtaagtac 5100
ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc aacaacgagt 5160
ccagcgtctg caaaacgtcg tcgtcagcga agtgcaagcg tcgcgtatcg gtgattcatt 5220
ctgctaacca gtaaggcaac cccgccagcc agcgcatagc cactaagtaa gacgattggt 5280
cattccgttg gggcggtcgg tagccgggtc ctcaacgaca ggagcacgat catgcgcacc 5340
cgtggggccg atcggcccag gagttgctgt cctcgtgcta gtacgcgtgg gcaccccggc 5400
ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ggtacggccg 5460
ctattaccgg acgaagagcg gctttgcaaa ccaccgccct ccagtgacga aggcttgagc 5520
gagggcgtgc aagattccga ataccgcaag ggtcactgct tccgaactcg ctcccgcacg 5580
ttctaaggct tatggcgttc cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc 5640
tcgccgaaaa gctgtccggc tagtagcagc gcgaggtcgc tttcgccagg agcggctttt 5700
tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag actgggtctc 5760
gcgacggccg tggacaggat gctcaacgta ctatttcttc acagtcataa gtgcggcgac 5820
gatagtcatg ccccgcgccc accggaagga tgtcagtatt cacgccgctg ctatcagtac 5880
ggggcgcggg tggccttcct gctgactggg ttgaaggctc tcaagggcat cggtcgagat 5940
cccggtgcct cgactgaccc aacttccgag agttcccgta gccagctcta gggccacgga 6000
aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc ttactcactc 6060
gattgaatgt aattaacgca acgcgagtga cgggcgaaag cagtcgggaa acctgtcgtg 6120
ccagctgcat taatgaatcg gccaacgcgc gtcagccctt tggacagcac ggtcgacgta 6180
attacttagc cggttgcgcg ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt 6240
tcttttcacc cccctctccg ccaaacgcat aacccgcggt cccaccaaaa agaaaagtgg 6300
agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag tcactctgcc 6360
cgttgtcgac taacgggaag tggcggaccg ggactctctc ttgcagcaag cggtccacgc 6420
tggtttgccc cagcaggcga aaatcctgtt aacgtcgttc gccaggtgcg accaaacggg 6480
gtcgtccgct tttaggacaa tgatggtggt taacggcggg atataacatg agctgtcttc 6540
ggtatcgtcg actaccacca attgccgccc tatattgtac tcgacagaag ccatagcagc 6600
tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat atagggtgat 6660
ggctctatag gcgtggttgc gcgtcgggcc tgagccatta ggcgcgcatt gcgcccagcg 6720
ccatctgatc gttggcaacc agcatcgcag ccgcgcgtaa cgcgggtcgc ggtagactag 6780
caaccgttgg tcgtagcgtc tgggaacgat gccctcattc agcatttgca tggtttgttg 6840
aaaaccggac acccttgcta cgggagtaag tcgtaaacgt accaaacaac ttttggcctg 6900
atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg taccgtgagg 6960
tcagcggaag ggcaaggcga tagccgactt aaactaacgc agtgagatat ttatgccagc 7020
cagccagacg cagacgcgcc gagacagaac tcactctata aatacggtcg gtcggtctgc 7080
gtctgcgcgg ctctgtcttg ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa 7140
tgcgaccaga aattacccgg gcgattgtcg cgctaaacga ccactgggtt acgctggtct 7200
tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt acgaggtgcg 7260
ggtcagcgca tggcagaagt accctctttt attatgacaa gatgggtgtc tggtcagaga 7320
catcaagaaa taacgccgga acattagtgc ctacccacag accagtctct gtagttcttt 7380
attgcggcct tgtaatcacg aggcagcttc cacagcaatg gcatcctggt catccagcgg 7440
atagttaatg tccgtcgaag gtgtcgttac cgtaggacca gtaggtcgcc tatcaattac 7500
atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca tagtcgggtg 7560
actgcgcaac gcgctcttct aacacgtggc ggcgaaatgt ggcttcgacg ccgcttcgtt 7620
ctaccatcga caccaccacg ctggcaccca ccgaagctgc ggcgaagcaa gatggtagct 7680
gtggtggtgc gaccgtgggt gttgatcggc gcgagattta atcgccgcga caatttgcga 7740
cggcgcgtgc caactagccg cgctctaaat tagcggcgct gttaaacgct gccgcgcacg 7800
agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc tcccggtctg 7860
acctccaccg ttgcggttag tcgttgctga caaacgggcg cagttgttgt gccacgcggt 7920
tgggaatgta attcagctcc gccatcgccg gtcaacaaca cggtgcgcca acccttacat 7980
taagtcgagg cggtagcggc cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc 8040
ctggttcacc gaaggtgaaa aagggcgcaa aagcgtcttt gcaccgaccg gaccaagtgg 8100
acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta tgcgcccttt 8160
gccagactat tctctgtggc cgtatgagac gctgtagcat taacgttact ggtttcacat 8220
tcaccaccct gaattgactc tcttccgggc attgcaatga ccaaagtgta agtggtggga 8280
cttaactgag agaaggcccg gctatcatgc cataccgcga aaggttttgc gccattcgat 8340
ggtgtccggg cgatagtacg gtatggcgct ttccaaaacg cggtaagcta ccacaggccc 8400
atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta tagagctgcg 8460
agagggaata cgctgaggac gtaatccttc gtcgggtcat gtaggttgag gccgttgagc 8520
accgccgccg caaggaatgg tgcatgcaag catccaactc cggcaactcg tggcggcggc 8580
gttccttacc acgtacgttc gagatggcgc ccaacagtcc cccggccacg gggcctgcca 8640
ccatacccac ctctaccgcg ggttgtcagg gggccggtgc cccggacggt ggtatgggtg 8700
gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggctttgtt 8760
cgcgagtact cgggcttcac cgctcgggct agaaggggta cggtgatgtc ggcgatatag 8820
gcgccagcaa ccgcacctgt ggcgccggtg gccactacag ccgctatatc cgcggtcgtt 8880
ggcgtggaca ccgcggccac atgccggcca cgatgcgtcc ggcgtagagg atcgagatct 8940
cgatcccgcg tacggccggt gctacgcagg ccgcatctcc tagctctaga gctagggcgc 9000
aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tttaattatg 9060
ctgagtgata tccccttaac actcgcctat tgttaagggg tctagaaata attttgttta 9120
actttaagaa ggagatatac atatgttccc agatctttat taaaacaaat tgaaattctt 9180
cctctatatg tatacaaggg gaccattccg ttatcccgtc tttttgacaa cgctatgctc 9240
cgtgcccatc ctggtaaggc aatagggcag aaaaactgtt gcgatacgag gcacgggtag 9300
gtctgcacca gctggccttt gacacctacc aggagtttga agaagcctat cagacgtggt 9360
cgaccggaaa ctgtggatgg tcctcaaact tcttcggata atcccaaagg aacagaagta 9420
ttcattcctg cagaaccccc agacctccct tagggtttcc ttgtcttcat aagtaaggac 9480
gtcttggggg tctggaggga ctgtttctca gagtctattc cgacaccctc caacagggag 9540
gaaacacaac gacaaagagt ctcagataag gctgtgggag gttgtccctc ctttgtgttg 9600
agaaatccaa cctagagctg ctccgcatct ccctgctgct catccagtcg tctttaggtt 9660
ggatctcgac gaggcgtaga gggacgacga gtaggtcagc tggctggagc ccgtgcagtt 9720
cctcaggagt gtcttcgcca acagcctggt accgacctcg ggcacgtcaa ggagtcctca 9780
cagaagcggt tgtcggacca gtacggcgcc tctgacagca acgtctatga cctcctaaag 9840
gacctagagg catgccgcgg agactgtcgt tgcagatact ggaggatttc ctggatctcc 9900
aaggcatcca aacgctgatg gggaggctgg aagatggcag cccccggact ttccgtaggt 9960
ttgcgactac ccctccgacc ttctaccgtc gggggcctga gggcagatct tcaagcagac 10020
ctacagcaag ttcgacacaa actcacacaa cccgtctaga agttcgtctg gatgtcgttc 10080
aagctgtgtt tgagtgtgtt cgatgacgca ctactcaaga actacgggct gctctactgc 10140
ttcaggaagg gctactgcgt gatgagttct tgatgcccga cgagatgacg aagtccttcc 10200
acatggacaa ggtcgagaca ttcctgcgca tcgtgcagtg ccgctctgtg tgtacctgtt 10260
ccagctctgt aaggacgcgt agcacgtcac ggcgagacac gagggcagct gtggcttcta 10320
aggatccgaa ttcgagctcc gtcgacaagc ctcccgtcga caccgaagat tcctaggctt 10380
aagctcgagg cagctgttcg ttgcggccgc actcgatgag caataactag cataacccct 10440
tggggcctct aacgccggcg tgagctactc gttattgatc gtattgggga accccggaga 10500
aaacgggtct tgaggggttt tttgctgaaa ggaggaattt gcccagaact ccccaaaaaa 10560
cgactttcct cctt 10574
Claims (99)
1.一种修饰的生长激素多肽,包含:
与未修饰的生长激素多肽相比,在相应于成熟人类生长激素多肽的氨基酸位置1-55、57、58、60-63、67-87、89-91、93、95-100、102-128、131-132、135-139、141、142、144、148-182、184、185和187-191的任何位置的1至5个氨基酸置换,其中:
成熟人类生长激素多肽包含SEQ ID NO:1的氨基酸序列;以及
所述修饰的生长激素多肽与SEQ ID NO:1或712的未修饰的生长激素多肽相比展示增加的蛋白质稳定性。
2.一种修饰的生长激素多肽,包含:
与未修饰的生长激素多肽相比,在相应于成熟人类生长激素多肽的氨基酸位置1-12、14-26、29-53、57、58、60-63、67-78、80-84、86、87、89、91、93、95-100、102-113、115-128、131、132、135-139、141、142、144、148-160、162-182、185和187-191的任何位置的一或多个单氨基酸置换,其中:
成熟人类生长激素多肽包含SEQ ID NO:1的氨基酸序列;以及
所述修饰的生长激素多肽与SEQ ID NO:1或712的未修饰的成熟人类生长激素多肽相比展示增加的蛋白质稳定性。
3.一种修饰的生长激素多肽,包含:
与未修饰的生长激素多肽相比一或多个单氨基酸置换,其中:
置换位置不是相应于包含SEQ ID NO:1的氨基酸序列的成熟人类生长激素多肽的位置56、59、64-66、88、92、94、101、129、130、133、134、140、143、145-147、183和186的位置;以及
所述修饰的生长激素多肽与未修饰的生长激素相比展示增加的蛋白质稳定性。
4.一种修饰的生长激素多肽,包含:
与未修饰的生长激素多肽相比一或多个单氨基酸置换,其中:
所述氨基酸置换相应于选自包含SEQ ID NO:1的氨基酸序列的成熟人类生长激素多肽的位置1-55、57、58、60-63、67-87、89-91、93、95-100、102-128、131、132、135-139、141、142、144、148-182、184、185和187-191的位置,其中:
如果位置9被置换,则置换氨基酸不是脯氨酸;
如果位置14被置换,则置换氨基酸不是丝氨酸;
如果位置13或27被置换,则置换氨基酸不是缬氨酸;
如果位置28被置换,则置换氨基酸不是苯丙氨酸;
如果位置54被置换,则置换氨基酸不是酪氨酸;
如果位置55、79、85或184被置换,则置换氨基酸不是丙氨酸;
如果位置90被置换,则置换氨基酸不是异亮氨酸;
如果位置114或161被置换,则置换氨基酸不是甲硫氨酸;以及
如果位置120或126被置换,则置换氨基酸不是精氨酸;以及
所述修饰的生长激素多肽与未修饰的生长激素多肽相比展示增加的蛋白质稳定性。
5.权利要求1或4的修饰的生长激素多肽,其中:
所述氨基酸置换相应于选自成熟人类生长激素的位置1、2、5、6、8、9、10、11、14、15、16、19、20、23、25、26、28、30、31、32、33、35、37、38、39、41、42、44、45、48、52、54、61、70、73、74、75、76、77、80、81、82、86、87、89、93、97、103、107、111、112、113、114、115、116、117、118、119、124、125、127、128、139、153、154、156、157、158、160、162、163、164、166、167、168、169、170、171、172、174、176、177、178和191的位置。
6.权利要求5的修饰的生长激素多肽,其中:
所述氨基酸置换相应于选自成熟人类生长激素多肽的位置F1、P2、P5、L6、R8、L9、F10、D11、M14、L15、R16、R19、L20、L23、F25、D26、Y28、E30、F31、E32、E33、Y35、P37、K38、E39、K41、Y42、F44、L45、P48、L52、F54、P61、K70、L73、E74、L75、L76、R77、L80、L81、L82、W86、L87、P89、L93、F97、Y103、D107、Y111、D112、L113、L114、K115、D116、L117、E118、E119、L124、M125、R127、L128、F139、D153、D154、L156、L157、K158、Y160、L162、L163、Y164、F166、R167、K168、D169、M170、D171、K172、E174、F176、L177、R178和F191的位置。
7.权利要求1-6任一项的修饰的生长激素多肽,其中:
所述氨基酸置换选自由H或Q置换R、由H、Q或N置换E、由Q或N置换K、由N或Q置换D、由I或V置换M、由A或S置换P、由I或H置换Y、由I或V置换F、由H或S置换W以及由I或V置换L。
8.权利要求5-7任一项的修饰的生长激素多肽,其中:
所述氨基酸置换选自F1I、F1V、P2A、P2S、P5A、P5S、L6I、L6V、R8H、R8Q、L9I、L9V、F10I、F10V、D11N、D11Q、M14I、M14V、L15I、L15V、R16H、R16Q、R19H、R19Q、L20I、L20V、L23I、L23V、F25I、F25V、D26N、D26Q、Y28H、Y28I、E30Q、E30H、E30N、F31I、F31V、E32Q、E32H、E32N、E33Q、E33H、E33N、Y35H、Y35I、P37A、P37S、K38N、K38Q、E39Q、E39H、E39N、K41N、K41Q、Y42H、Y42I、F44I、F44V、L45I、L45V、P48A、P48S、L52I、L52V、F54I、F54V、P61A、P61S、K70N、K70Q、L73I、L73V、E74Q、E74H、E74N、L75I、L75V、L76I、L76V、R77H、R77Q、L80I、L80V、L81I、L81V、L82I、L82V、W86H、W86S、L87I、L87V、P89A、P89S、L93I、L93V、F97I、F97V、Y103H、Y103I、D107N、D107Q、Y111H、Y111I、D112N、D112Q、L113I、L113V、L114I、L114V、K115N、K115Q、D116N、D116Q、L117I、L117V、E118Q、E118H、E118N、E119Q、E119H、E119N、L124I、L124V、M125I、M125V、R127H、R127Q、L128I、L128V、F139I、F139V、D153N、D153Q、D154N、D154Q、L156I、L156V、L157I、L157V、K158N、K158Q、Y160H、Y160I、L162I、L162V、L163I、L163V、Y164H、Y164I、F166I、F166V、R167H、R167Q、K168N、K168Q、D169N、D169Q、M170I、M170V、D171N、D171Q、K172N、K172Q、E174Q、E174H、E174N、F176I、F176V、L177I、L177V、R178H、R178Q、F191I和F191V。
9.权利要求1-4任一项的修饰的生长激素多肽,其中:
所述氨基酸置换相应于选自成熟人类生长激素多肽的位置1、2、5、9、11、14、16、23、26、38、41、42、73、74、81、87、111、112、116、119、124、125、153、156、157、158、162、166、167、168、169、171、172、174、177、178和191的位置。
10.权利要求1-4任一项的修饰的生长激素多肽,其中:
所述氨基酸置换相应于选自成熟人类生长激素多肽的位置F1、P2、P5、L9、D11、M14、R16、L23、D26、K38、K41、Y42、L73、E74、L81、L87、Y111、D112、D116、E119、L124、M125、D153、L156、L157、K158、L162、F166、R167、K168、D169、D171、D171、K172、E174、L177、R178和F191的位置。
11.权利要求10的修饰的生长激素多肽,其中:
所述氨基酸置换选自F1I、P2A、P5S、L9V、D11N、M14V、R16H、L23I、L23V、D26N、K38N、K41Q、Y42H、Y42I、L73V、E74N、L81V、L87V、Y111I、D112N、D116Q、E119Q、L124V、M125I、M125V、D153N、L156I、L157I、K158N、L162I、F166I、R167H、R167Q、K168N、K168Q、D169Q、D171N、D171Q、K172Q、E174Q、E174N、E174H、L177V、L177I、R178Q和F191I。
12.权利要求1或4的修饰的生长激素,其中:
所述氨基酸置换相应于选自成熟人类生长激素多肽的位置6、9、13、15、17、20、23、24、105、110、113、114、117、121、124和128的位置。
13.权利要求1或4的修饰的生长激素多肽,其中所述单氨基酸置换相应于选自成熟人类生长激素多肽的位置L6、L9、A13、L15、A17、L20、L23、A24、A105、V110、L113、L114、L117、I121、L124和L128的位置。
14.权利要求13的修饰的生长激素多肽,其中所述一或多个单氨基酸置换选自E、D、K、R、N、Q、S和T。
15.权利要求14的修饰的生长激素多肽,其中:
所述单氨基酸置换选自L6E、L6D、L6K、L6R、L6N、L6Q、L6S、L6T、L9E、L9D、L9K、L9R、L9N、L9Q、L9S、L9T、A13E、A13D、A13K、A13R、A13N、A13Q、A13S、A13T、L15E、L15D、L15K、L15R、L15N、L15Q、L15S、L15T、A17E、A17D、A17K、A17R、A17N、A17Q、A17S、A17T、L20E、L20D、L20K、L20R、L20N、L20Q、L20S、L20T、L23E、L23D、L23K、L23R、L23N、L23Q、L23S、L23T、A24E、A24D、A24K、A24R、A24N、A24Q、A24S、A24T、A105E、A105D、A105K、A105R、A105N、A105Q、A105S、A105T、V110E、V110D、V110K、V110R、V110N、V110Q、V110S、V110T、L113E、L113D、L113K、L113R、L113N、L113Q、L113S、L113T、L114E、L114D、L114K、L114R、L114N、L114Q、L114S、L114T、L117E、L117D、L117K、L117R、L117N、L117Q、L117S、L117T、I121E、I121D、I121K、I121R、I121N、I121Q、I121S、I121T、L124E、L124D、L124K、L124R、L124N、L124Q、L124S、L124T、L128E、L128D、L128K、L128R、L128Q、L128N、L128S和L128T。
16.权利要求1-11任一项的修饰的生长激素多肽,进一步包含:
相应于选自成熟人类生长激素多肽的位置6、9、13、15、17、20、23、24、105、110、113、114、117、121、124和128的位置的一或多个单氨基酸置换,其中:
如果位置13被置换,则置换氨基酸不是缬氨酸;以及
如果位置114被置换,则置换氨基酸不是甲硫氨酸。
17.权利要求16的修饰的生长激素多肽,其中:
所述一或多个单氨基酸置换相应于选自位置L6、L9、A13、L15、A17、L20、L23、A24、A105、V110、L113、L114、L117、I121、L124和L128的一或多个位置。
18.权利要求16或17的修饰的生长激素多肽,其中所述一或多个氨基酸置换选自E、D、K、R、N、Q、S和T。
19.权利要求16-18任一项的修饰的生长激素多肽,其中:
所述一或多个单氨基酸置换选自L6E、L6D、L6K、L6R、L6N、L6Q、L6S、L6T、L9E、L9D、L9K、L9R、L9N、L9Q、L9S、L9T、A13E、A13D、A13K、A13R、A13N、A13Q、A13S、A13T、L15E、L15D、L15K、L15R、L15Q、L15N、L15S、L15T、A17E、A17D、A17K、A17R、A17N、A17Q、A17S、A17T、L20E、L20D、L20K、L20R、L20N、L20Q、L20S、L20T、L23E、L23D、L23K、L23R、L23N、L23Q、L23S、L23T、A24E、A24D、A24K、A24R、A24N、A24Q、A24S、A24T、A105E、A105D、A105K、A105R、A105N、A105Q、A105S、A105T、V110E、V110D、V110K、V110R、V110N、V110Q、V110S、V110T、L113E、L113D、L113K、L113R、L113N、L113Q、L113S、L113T、L114E、L114D、L114K、L114R、L114N、L114Q、L114S、L114T、L117E、L117D、L117K、L117R、L117N、L117Q、L117S、L117T、I121E、I121D、I121K、I121R、I121N、I121Q、I121S、I121T、L124E、L124D、L124K、L124R、L124N、L124Q、L124S、L124T、L128E、L128D、L128K、L128R、L128Q、L128N、L128S和L128T。
20.权利要求2-20任一项的修饰的生长激素多肽,其中:
与未修饰的生长激素相比被置换的位置数是2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19或20个。
21.权利要求1-20任一项的修饰的生长激素多肽,进一步包含:
相应于选自包含SEQ ID NO:1的氨基酸序列的成熟人类生长激素多肽的位置56、59、64、65、66、88、92、94、101、129、130、133、134、140、143、145、146、147、183和186的一或多个单氨基酸置换。
22.权利要求21的修饰的生长激素多肽,其中所述氨基酸置换相应于选自位置E56、P59、R64、E65、E66、E88、F92、R94、L101、E129、D130、P133、R134、K140、Y143、K145、F146、D147、R183和E186的一或多个位置。
23.权利要求21或22的修饰的生长激素多肽,其中:
所述一或多个氨基酸置换选自用Q、N和H的任一个置换E、用S或A置换P、用H或Q置换R、用I或V置换L或F、用Q或N置换K或D以及用H或L置换Y。
24.权利要求21-23任一项的修饰的生长激素多肽,其中:
所述一或多个单氨基酸置换选自E56Q、E56N、E56H、P59S、P59A、R64H、R64Q、E65Q、E65N、E65H、E66Q、E66N、E66H、E88Q、E88N、E88H、F92I、F92V、R94H、R94Q、L101V、L101I、E129Q、E129N、E129H、D130Q、D130N、P133S、P133A、R134H、R134Q、K140Q、K140N、Y143H、Y143I、K145Q、K145N、F146I、F146V、D147Q、D147N、R183H、R183Q、E186Q、E186N和E186H。
25.权利要求1-24任一项的修饰的生长激素多肽,其是人类生长激素(hGH)多肽。
26.权利要求1-25任一项的修饰的生长激素多肽,其中所述氨基酸置换是在人垂体生长激素多肽或人胎盘生长激素多肽中进行。
27.权利要求1-24任一项的修饰的生长激素多肽,其是非人生长激素多肽。
28.权利要求1-27任一项的修饰的生长激素多肽,其是成熟生长激素多肽。
29.权利要求1-27任一项的修饰的生长激素多肽,其是前体生长激素多肽。
30.权利要求1-29任一项的修饰的生长激素多肽,其长度为191、192、193、194、195、196、197、198、199、200、201、202、203、204、205、206、207、208、209、210、211、212、213、214、215、216、217、218、219、220、221或222个氨基酸。
31.权利要求1-30任一项的修饰的生长激素多肽,其中:
所述一或多个氨基酸置换选自天然氨基酸、非天然氨基酸以及天然和非天然氨基酸的组合。
32.权利要求1-31任一项的修饰的生长激素多肽,其是裸多肽链。
33.权利要求1-31任一项的修饰的生长激素多肽,其中所述生长激素是聚乙二醇化的、白蛋白化的或糖基化的。
34.权利要求1-33任一项的修饰的生长激素多肽,进一步包含一或多个假野生型突变。
35.权利要求34的修饰的生长激素多肽,其中:
所述假野生型突变选自未修饰的生长激素的氨基酸残基的一或多个插入、缺失和置换。
36.权利要求1-35任一项的修饰的生长激素多肽,其中增加的稳定性表现为增加的对蛋白酶解的抗性。
37.权利要求36的修饰的生长激素多肽,其中所述增加的对蛋白酶解的抗性发生在血清、血液、唾液、消化液中或体外暴露于蛋白酶时。
38.权利要求36的修饰的生长激素多肽,其中所述增加的对蛋白酶解的抗性由所述的修饰的生长激素被口服给药或存在于消化道中时展示。
39.权利要求1-38任一项的修饰的生长激素多肽,其中与未修饰的生长激素相对比,所述增加的对蛋白酶解的抗性包括对一或多种蛋白酶的增加的抗性,所述蛋白酶选自胰蛋白酶、胰蛋白酶(Arg blocked)、胰蛋白酶(Lys blocked)、梭菌蛋白酶、内切蛋白酶Asp-N、胰凝乳蛋白酶、溴化氰、iodozobenzoate、Myxobacter P.、Armillaria、鲁米那胃蛋白酶、微绒毛内肽酶、二肽基肽酶、肠肽酶、水解酶、明胶酶B和明胶酶A。
40.权利要求1-35任一项的修饰的生长激素多肽,其中所述增加的稳定性表现为增加的热耐受性。
41.权利要求40的修饰的生长激素多肽,其在约20℃至约45℃的温度下具有增加的热耐受性。
42.权利要求40的修饰的生长激素多肽,其在对象的体温下具有增加的热耐受性。
43.权利要求40的修饰的生长激素多肽,其在约37℃具有增加的热耐受性。
44.权利要求1-43任一项的修饰的生长激素多肽,其中所述增加的稳定性表现为增加的体内或体外半衰期。
45.权利要求1-44任一项的修饰的生长激素多肽,其中所述增加的稳定性表现为当被给予对象时的增加的半衰期。
46.权利要求1-45任一项的修饰的生长激素多肽,其中所述修饰的生长激素的半衰期与未修饰的生长激素的半衰期相比增加量为至少10%、至少20%、至少30%、至少40%、至少50%、至少60%、至少70%、至少80%、至少90%、至少100%、至少150%、至少200%、至少250%、至少300%、至少350%、至少400%、至少450%或至少500%。
47.权利要求1-45任一项的修饰的生长激素多肽,其中所述修饰的生长激素的半衰期与未修饰的生长激素多肽的半衰期相比增加量为至少6倍、7倍、8倍、9倍、10倍、20倍、30倍、40倍、50倍、60倍、70倍、80倍、90倍、100倍、200倍、300倍、400倍、500倍、600倍、700倍、800倍、900倍或1000倍或更多倍。
48.权利要求1-47任一项的修饰的生长激素多肽,其与未修饰的生长激素相比具有增加的生物学活性。
49.权利要求1-47任一项的修饰的生长激素多肽,其与未修饰的生长激素相比具有降低的生物学活性。
50.权利要求48或49的修饰的生长激素多肽,其中生物学活性通过测量体外细胞增殖而评价。
51.权利要求48或49的修饰的生长激素多肽,其中所述修饰的生长激素多肽结合生长激素受体。
52.权利要求1-37或44-51任一项的修饰的生长激素多肽,其中:
所述修饰的生长激素多肽与未修饰的生长激素多肽相比展示增加的对蛋白酶解的抗性并展示降低的热耐受性。
53.权利要求1-35或40-51任一项的修饰的生长激素多肽,其中:
所述修饰的生长激素多肽与未修饰的生长激素多肽相比展示增加的热耐受性并展示降低的对蛋白酶解的抗性。
54.一种文库,其包含2种或多种权利要求1-53任一项的修饰的生长激素多肽。
55.一种核酸分子,其包含编码权利要求1-53任一项的修饰的生长激素多肽的核苷酸序列。
56.一种核酸分子文库,其包含权利要求55的多个分子。
57.一种载体,其包含权利要求55的核酸分子。
58.一种真核细胞,其包含权利要求57的载体。
59.一种原核细胞,其包含权利要求57的载体。
60.一种文库,其包含多个权利要求57的载体。
61.一种用于表达修饰的生长激素多肽的方法,包括:
i)将权利要求55的核酸或权利要求57的载体导入细胞中,以及
ii)在被编码的修饰的生长激素多肽表达的条件下培养所述细胞。
62.权利要求61的方法,其中所述细胞是真核细胞。
63.权利要求61或62的方法,其中所述修饰的生长激素多肽是糖基化的。
64.一种修饰的生长激素多肽,其包含SEQ ID NO:2-69、75、76、85-107、111、112、115、116、119、120、123-154、164、165、176-216、222-351所示的任一氨基酸序列,或其生物学活性部分。
65.一种药物组合物,其包含权利要求1-53和64任一项的修饰的生长激素多肽。
66.权利要求65的药物组合物,还包含药物学可接受的赋形剂。
67.权利要求65的药物组合物,其中所述药物组合物被配制用于口服、鼻或肺给药。
68.权利要求65的药物组合物,其中所述药物组合物被配制用于口服给药。
69.权利要求68的药物组合物,其中所述修饰的生长激素与未修饰的生长激素相比在如下条件下展示增加的半衰期,所述条件选自暴露于唾液、暴露于胃肠道中的蛋白酶和暴露于低pH条件。
70.权利要求65的药物组合物,其中所述修饰包括去除蛋白酶消化位点或增加蛋白质结构的稳定性。
71.权利要求65-70任一项的药物组合物,其中所述修饰的生长激素多肽展示在胃肠道中的增加的蛋白质半衰期或生物利用度。
72.权利要求71的药物组合物,其中蛋白质半衰期与未修饰的生长激素的半衰期相比增加量为至少10%、至少20%、至少30%、至少40%、至少50%、至少60%、至少70%、至少80%、至少90%、至少100%、至少150%、至少200%、至少250%、至少300%、至少350%、至少400%、至少450%或至少500%或更多。
73.权利要求71的药物组合物,其中蛋白质半衰期与未修饰的生长激素的半衰期相比增加量为至少6倍、7倍、8倍、9倍、10倍、20倍、30倍、40倍、50倍、60倍、70倍、80倍、90倍、100倍、200倍、300倍、400倍、500倍、600倍、700倍、800倍、900倍或1000倍或更多倍。
74.权利要求71的药物组合物,其中所述修饰的生长激素的生物学活性与未修饰的生长激素相比是增加的。
75.权利要求71-74任一项的药物组合物,其中蛋白质半衰期在暴露于选自如下的一或多种条件后是增加的,所述条件选自暴露于唾液、暴露于胃肠道中的蛋白酶和暴露于低pH条件。
76.权利要求67的药物组合物,其中不使用蛋白酶抑制剂而制备所述药物组合物。
77.权利要求76的药物组合物,其中所述蛋白酶抑制剂选自Bowman-Birk抑制剂、缀合的Bowman-Birk抑制剂、抑酶肽和卡莫他特。
78.权利要求66的药物组合物,其中所述赋形剂选自粘合剂、填充剂、润滑剂、崩解剂和湿润剂。
79.权利要求68的药物组合物,其中所述药物组合物配制为以液体、丸剂、片剂或胶囊剂形式给药。
80.权利要求79的药物组合物,其中所述丸剂或片剂是可咀嚼的。
81.权利要求79的药物组合物,其中所述胶囊中的药物组合物呈液体形式。
82.权利要求79的药物组合物,其中所述液体选自溶液、糖浆和悬浮液。
83.权利要求67的药物组合物,其中所述药物组合物被配制用于所述修饰的生长激素多肽的控制释放。
84.权利要求83的药物组合物,其中所述组合物呈片剂或锭剂形式。
85.权利要求84的药物组合物,其中所述锭剂将所述修饰的生长激素多肽输送至口腔粘膜、喉粘膜或胃肠道。
86.权利要求84的药物组合物,其中所述锭剂用赋形剂配制。
87.权利要求83的药物组合物,其中所述赋形剂选自无水结晶麦芽糖和硬脂酸镁。
88.权利要求65-87任一项的药物组合物,其中所述药物组合物不加保护性化合物而配制。
89.权利要求88的药物组合物,其中所述修饰的生长激素展示对胃肠道蛋白酶的抗性。
90.权利要求65-89任一项的药物组合物,进一步包含药物学可接受的添加剂。
91.权利要求90的药物组合物,其中所述添加剂选自悬浮剂、乳化剂、非水载体和防腐剂。
92.一种药物组合物,其包含权利要求55的核酸分子或权利要求57的载体。
93.一种制品,其包括包装材料和含在所述包装材料中的权利要求65-92任一项的药物组合物。
94.权利要求96的制品,其中所述的修饰的生长激素有效治疗生长激素疾病或病症,所述包装材料包括指示所述修饰的生长激素用于治疗生长激素疾病或病症的标签。
95.一种试剂盒,其包含权利要求65-92任一项的药物组合物、用于给予修饰的GH多肽的装置和任选地给药指导和试剂。
96.权利要求65-92任一项的药物组合物在制备用于治疗患有经给予生长激素可以治疗的疾病或状况的对象的药物的应用。
97.权利要求1-53和64任一项的生长激素多肽在制备用于治疗患有经给予生长激素可以治疗的疾病或状况的对象的药物中的应用。
98.权利要求96或97的应用,其中所述疾病或状况选自生长缺陷疾病、AIDS消瘦、衰老、HIV感染对象的受损的免疫功能、分解代谢疾病、手术恢复、充血性心肌病、肝移植、肝切除术后肝再生、慢性肾功能衰竭、肾性骨营养不良、骨质疏松、软骨发育不全/季肋发育不全、骨骼发育异常、慢性炎性或营养性疾病如克罗恩氏病、短肠综合征、少年慢性关节炎、囊性纤维化、男性不育症、X连锁低磷血症性佝偻病、唐氏综合症、脊柱裂、努南综合征、肥胖症、肌肉强度受损以及纤维肌痛。
99.权利要求98的应用,其中所述生长缺陷疾病选自特纳氏综合征、子宫内生长迟缓、特发性身材矮小症、Prader Willi综合征和地中海贫血。
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- 2005-11-04 WO PCT/IB2005/003662 patent/WO2006048777A2/en active Application Filing
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
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CN101343624A (zh) * | 2008-06-25 | 2009-01-14 | 浙江中奇生物药业股份有限公司 | 一种重组人生长激素基因的杆状病毒及其制备方法与应用 |
CN102753573A (zh) * | 2009-12-21 | 2012-10-24 | Ambrx公司 | 经过修饰的牛促生长素多肽和其用途 |
CN107674121A (zh) * | 2009-12-21 | 2018-02-09 | Ambrx 公司 | 经过修饰的牛促生长素多肽和其用途 |
CN110448683A (zh) * | 2011-12-09 | 2019-11-15 | 麦特保利药业有限公司 | 生长激素片段的用途 |
Also Published As
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JP2011097944A (ja) | 2011-05-19 |
JP2008518615A (ja) | 2008-06-05 |
WO2006048777A3 (en) | 2006-09-28 |
CA2586389A1 (en) | 2006-05-11 |
US20060094655A1 (en) | 2006-05-04 |
KR100994610B1 (ko) | 2010-11-15 |
US20060247170A1 (en) | 2006-11-02 |
KR20070092218A (ko) | 2007-09-12 |
US8222209B2 (en) | 2012-07-17 |
RU2007120182A (ru) | 2008-12-10 |
JP5017119B2 (ja) | 2012-09-05 |
CN101094865B (zh) | 2012-04-25 |
EP2241574A1 (en) | 2010-10-20 |
EP2476700A1 (en) | 2012-07-18 |
AU2005300257A1 (en) | 2006-05-11 |
US20080026993A9 (en) | 2008-01-31 |
US20110130331A1 (en) | 2011-06-02 |
WO2006048777A2 (en) | 2006-05-11 |
US7998930B2 (en) | 2011-08-16 |
EP1807449A2 (en) | 2007-07-18 |
US20070249532A9 (en) | 2007-10-25 |
JP5117563B2 (ja) | 2013-01-16 |
EP2241574B1 (en) | 2015-02-25 |
US7884073B2 (en) | 2011-02-08 |
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