CN108602869A - Fgf21变体 - Google Patents

Fgf21变体 Download PDF

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CN108602869A
CN108602869A CN201680070380.3A CN201680070380A CN108602869A CN 108602869 A CN108602869 A CN 108602869A CN 201680070380 A CN201680070380 A CN 201680070380A CN 108602869 A CN108602869 A CN 108602869A
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M·索末菲尔德
T·兰格
O·博沙伊嫩
M·德雷尔
W·迪特里希
P·哈贝曼
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Sanofi SA
Sanofi Aventis France
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Abstract

本发明涉及人成纤维细胞生长因子21(FGF21)的多肽变体及其融合分子，以及编码上述的核酸分子。其还涉及它们作为药物的用途，尤其是用于治疗肥胖症、超重、代谢综合征、糖尿病、高血糖症、血脂异常、非酒精性脂肪性肝炎(NASH)和/或动脉粥样硬化。

Description

FGF21变体
发明的技术领域
本发明涉及人成纤维细胞生长因子21(FGF21)的多肽变体及其融合分子，以及编码上述的核酸分子。其进一步涉及它们作为药物的用途，尤其是用于治疗肥胖症，超重，代谢综合征，糖尿病，高血糖症，血脂异常，非酒精性脂肪性肝炎(NASH)和/或动脉粥样硬化。
发明的背景技术
成纤维细胞生长因子(FGF)是在发育和成熟组织中广泛表达的多肽。该FGF家族目前由FGF-1到FGF-23的22个成员组成(其中一个称为FGF15/19)。FGF家族成员在脊椎动物物种间的基因结构和氨基酸序列高度保守。有18种哺乳动物成纤维细胞生长因子(FGF1-FGF10和FGF16-FGF23)，它们根据序列同源性和种系发生上的差异分为6个亚家族。编号的未分配到亚家族的“FGF”，即FGF同源因子(之前称为FGF11-FGF14)与FGF家族具有高度序列同一性，但不激活FGF受体(FGFR)，因此通常不被认为是FGF家族的成员。尽管大多数FGF充当细胞生长和分化的局部调节物，但最近研究指示包括FGF15/19、FGF21和FGF23在内的FGF19亚家族成员通过内分泌方式发挥重要的代谢作用。FGF19亚家族成员调控多种不受经典FGF影响的生理过程。这些内分泌因子种类广泛的代谢活动包括调控胆汁酸、碳水化合物和脂质代谢，以及磷酸盐、钙和维生素D体内稳态。
FGF21最初从小鼠胚胎中分离。FGF21 mRNA在肝脏中表达最丰富，在胸腺中表达程度较少。人FGF21与小鼠FGF21高度相似(约81％的氨基酸同一性)。在人FGF家族成员中，FGF21与FGF19最相似(约30％的氨基酸同一性)。FGF21没有表现出增殖和致瘤效应，而所述效应对FGF家族的大部分成员是很典型的。
成纤维细胞生长因子21(FGF21)是主要由肝脏产生的新型代谢调控因子，其在肥胖症和2型糖尿病动物模型中发挥有效的抗糖尿病和降脂作用。该激素有助于体重调节，并且涉及小鼠对营养缺乏(nutritional deprivation)和生酮状态(ketogenic state)的响应。FGF21代谢作用的主要位点是脂肪组织、肝脏和胰腺。实验研究已显示，在对糖尿病小鼠和灵长类施用FGF21后，糖尿病代偿和血脂异常有改善。已显示在存在和不存在胰岛素时FGF21刺激小鼠3T3-L1脂肪细胞中葡萄糖吸收，并以剂量依赖的方式减少ob/ob及db/db小鼠和8周龄的ZDF大鼠中的摄食和空腹血糖、甘油三酯和胰高血糖素水平，因此这为将FGF-21用作治疗糖尿病和肥胖症的疗法提供了基础。
将FGF21施用于肥胖瘦蛋白缺陷型ob/ob小鼠、瘦蛋白受体缺陷型db/db小鼠和肥胖ZDF大鼠显著地降低血糖和甘油三酯，减低空腹胰岛素水平，并且在口服葡萄糖耐量测试过程中提高葡萄糖清除率。在施用两周的过程中，FGF21没有影响糖尿病或瘦/去脂肪(lean)小鼠和大鼠的食物摄入或体重/构成。重要的是，FGF21在糖尿病或健康动物中测试的任何剂量下，或当在转基因小鼠中过表达时都没有诱导有丝分裂、低血糖或体重增加。FGF21过表达的转基因小鼠对饮食诱导的肥胖有抗性。
将FGF21施用于糖尿病恒河猴6周降低空腹血浆葡萄糖、果糖胺、甘油三酯、胰岛素和胰高血糖素水平。重要的是，尽管有显著的降低葡萄糖作用，但在研究中没有观察到低血糖症。施用FGF21也显著降低LDL胆固醇和增加HDL胆固醇，并且与小鼠不同的是，轻微但显著地降低了体重。
发明内容
与人野生型FGF21作为治疗剂用于治疗糖尿病、肥胖症、代谢综合征和其他适应症的相关问题在于其体内半衰期有限。在小鼠中FGF21具有低于1小时的半衰期，而在食蟹猴(Macaca fascicularis)中仅为约2-4小时。因此，在用作治疗剂的FGF21蛋白的开发中，需要相对于人野生型FGF21具有改进的药学特性的变体，尤其是增加针对蛋白酶和/或热降解的稳定性和/或增加的效力/功效。为了达到这个目标，将FGF21变体设计为识别具有增强的稳定性的FGF21多肽。
在第一个方面，本发明涉及人成纤维细胞生长因子21(FGF21)的变体，其包含SEQID NO：171的氨基酸序列，
其中
-Xaa55是Q或另一氨基酸，如G或C；
-Xaa147是P或C，或是缺失的；
-Xaa148是G或C，或是缺失的；
-Xaa149是N或C，或是缺失的；
-Xaa150是K或另一氨基酸，如H，或是缺失的；
-Xaa151是S或是缺失的；
-Xaa152是P或另一氨基酸，如A或L，或是缺失的；
-Xaa153是H或另一氨基酸，如Q，Y或K，或是缺失的；
-Xaa1 54是R或另一氨基酸，如Q或K，或是缺失的；
-Xaa155是D或另一氨基酸，如L，K，Y，P，E，N或C，或是缺失的；
-Xaa156是P或另一氨基酸，如A或K，或是缺失的；
-Xaa157是A或另一氨基酸，如V，G或S，或是缺失的；
-Xaa158是P或另一氨基酸，如H，或是缺失的；
-Xaa159是R或另一氨基酸，如H，K或Q，或是缺失的；
-Xaa160是G或是缺失的；
-Xaa161是P或C，或是缺失的；
-Xaa162是A或另一氨基酸，如C或Y，或是缺失的；
-Xaa163是R或另一氨基酸，如H，或是缺失的；
-Xaa184是Q或是缺失的；
-Xaa185是P或是缺失的；
-Xaa186是P或是缺失的；
-Xaa187是D或是缺失的；
-Xaa188是V或是缺失的；
-Xaa189是G或是缺失的；
-Xaa190是S或是缺失的；
-Xaa191是S或是缺失的；
-Xaa192是D或是缺失的；
-Xaa193是P或是缺失的；
-Xaa194是L或是缺失的；
-Xaa195是S或C，或是缺失的；
-Xaa196是M或另一氨基酸，如P，V或C，或是缺失的；
-Xaa197是V或另一氨基酸，如E，D，G或M，或是缺失的；
-Xaa198是G或另一氨基酸，如E，D，R，K，Y，P或V，或是缺失的；
-Xaa199是P或另一氨基酸，如S，Q，R，T，G，F，L，D或M，
或是缺失的；
-Xaa200是S或另一氨基酸，如Q，M，C，P，N或H，或是缺失的；
-Xaa201是Q或另一氨基酸，如P或S，或是缺失的；
-Xaa202是G或另一氨基酸，如T，或是缺失的；
-Xaa203是R或另一氨基酸，如E，H或C，或是缺失的；
-Xaa204是S或是缺失的；
-Xaa205是P或是缺失的；
-Xaa206是S或是缺失的；且
-Xaa207是Y或是缺失的；
其中，任选地，SEQ ID NO：171包含以C对如下氨基酸中至少一个的取代：R47，L49，T51，A54，Q56，A59，H60，E62，I63，G67，V69，G71，A72，A73，S76，P77，E78，S79，L80，L81，Q82，L83，I91，L94，G95，V96，K97，T98，R100，L102，Q104，D107，G108，L110，G112，L114，A120，R124，D130，Y132，Q136，S137，A139，H140，L142，P143，H145，L146，L165，L167，L170，P174；
其条件是SEQ ID NO：171不是成熟人野生型FGF21(SEQ ID NO：2)且与成熟人野生型FGF21(SEQ ID NO：2)相比包含0，2，4，6或8个额外的半胱氨酸，
其中，任选地，SEQ ID NO：171还包含突变G141S和/或突变P174L。
在一个实施方案中，
Xaa147至Xaa159被缺失并由蛋白酶抗性肽接头替代，如SEQ ID NO：161或SEQ IDNO：162的肽接头；
Xaa149至Xaa155被缺失并由蛋白酶抗性肽接头替代，如SEQ ID NO：161，SEQ IDNO：163或SEQ ID NO：164的肽接头；
Xaa149至Xaa159被缺失并由蛋白酶抗性肽接头替代，如SEQ ID NO：161或SEQ IDNO：162的肽接头；
Xaa149至Xaa162被缺失并由蛋白酶抗性肽接头替代，如SEQ ID NO：163或SEQ IDNO：165的肽接头；
Xaa149至Xaa163被缺失并由蛋白酶抗性肽接头替代，如SEQ ID NO：161的肽接头；
Xaa159至Xaa163被缺失并由蛋白酶抗性肽接头替代，如GA，GY，HH，GE或HE；
Xaa197至Xaa203被缺失并由蛋白酶抗性肽接头替代，如SEQ ID NO：166或SEQ IDNO：167；
蛋白酶抗性肽接头，如SEQ ID NO：168的肽接头，被插入Xaa198和Xaa199之间；和/或
SEQ ID NO：169或SEQ ID NO：170的氨基酸序列被添加于SEQ ID NO：171的S209之后。
在一个实施方案中，SEQ ID NO：171与成熟人野生型FGF21(SEQ ID NO：2)具有至少90％或至少91％或至少92％或至少93％或至少94％或至少95％或至少96％或至少97％或至少98％序列同一性。
在一个实施方案中，
-Xaa55是Q或C；
-Xaa147是P或C；
-Xaa148是G或C；
-Xaa149是N或C；
-Xaa150是K；
-Xaa151是S；
-Xaa152是P；
-Xaa153是H；
-Xaa154是R；
-Xaa155是D或C；
-Xaa156是P；
-Xaa157是A；
-Xaa158是P；
-Xaa159是R；
-Xaa160是G；
-Xaa161是P或C；
-Xaa162是A或C；
-Xaa163是R；
-Xaa184是Q；
-Xaa185是P；
-Xaa186是P；
-Xaa187是D；
-Xaa188是V；
-Xaa189是G；
-Xaa190是S；
-Xaa191是S；
-Xaa192是D；
-Xaa193是P；
-Xaa194是L；
-Xaa195是S或C；
-Xaa196是M或另一氨基酸，如P，V或C，或是缺失的；
-Xaa197是V或另一氨基酸，如E，D，G或M，或是缺失的；
-Xaa198是G或另一氨基酸，如E，D，R，K，Y，P或V，或是缺失的；
-Xaa199是P或另一氨基酸，如S，Q，R，T，G，F，L，D或M，或是缺失的；
-Xaa200是S或另一氨基酸，如Q，M，C，P，N或H，或是缺失的；
-Xaa201是Q或另一氨基酸，如P或S，或是缺失的；
-Xaa202是G或另一氨基酸，如T，或是缺失的；
-Xaa203是R或另一氨基酸，如E，H或C，或是缺失的；
-Xaa204是S；
-Xaa205是P；
-Xaa206是S；且
-Xaa207是Y；
其中，任选地，SEQ ID NO：171包含以C对如下氨基酸中至少一个的取代：R47，L49，T51，A54，Q56，A59，H60，E62，I63，G67，V69，G71，A72，A73，S76，P77，E78，S79，L80，L81，Q82，L83，I91，L94，G95，V96，K97，T98，R100，L102，Q104，D107，G108，L110，G112，L114，A120，R124，D130，Y132，Q136，S137，A139，H140，L142，P143，H145，L146，L165，L167，L170，P174。
在一个实施方案中，所述变体包含SEQ ID NO：175的氨基酸序列(其中变量如此处定义)，
其中，任选地，SEQ ID NO：175包含以C对如下氨基酸中至少一个的取代：R47，L49，T51，A54，Q56，A59，H60，E62，I63，G67，V69，G71，A72，A73，S76，P77，E78，S79，L80，L81，Q82，L83，I91，L94，G95，V96，K97，T98，R100，L102，Q104，D107，G108，L110，G112，L114，A120，R124，D130，Y132，Q136，S137，A139，H140，L142，P143，H145，L146，L165，L167，L170，P174；
其条件是SEQ ID NO：175不是成熟人野生型FGF21(SEQ ID NO：2)且与成熟人野生型FGF21(SEQ ID NO：2)相比包含0，2，4，6或8个额外的半胱氨酸，
其中，任选地，SEQ ID NO：175还包含突变G141S和/或突变P174L。
在一个实施方案中，
-Xaa55是Q或C；
-Xaa147是P或C；
-Xaa148是G；
-Xaa149是N或C；
-Xaa155是D或C；
-Xaa161是P；
-Xaa162是A；
-Xaa195是S；
-Xaa196是M；
-Xaa197是V；
-Xaa198是G或另一氨基酸，如E，D，R，K，Y，P或V，或是缺失的；
-Xaa199是P或另一氨基酸，如S，Q，R，T，G，F，L，D或M，或是缺失的；
-Xaa200是S；
-Xaa201是Q；
-Xaa202是G；且
-Xaa203是R；
其中，任选地，SEQ ID NO：171包含以C对如下氨基酸中至少一个的取代：R47，A59，H60，G71，S76，S79，D107，G108，L142，P174。
在一个实施方案中，所述变体包含SEQ ID NO：176的氨基酸序列(其中变量如此处定义)，
其中，任选地，SEQ ID NO：176包含以C对如下氨基酸中至少一个的取代：R47，A59，H60，G71，S76，S79，D107，G108，L142，P174；
其条件是SEQ ID NO：176不是成熟人野生型FGF21(SEQ ID NO：2)且与成熟人野生型FGF21(SEQ ID NO：2)相比包含0，2，4，6或8个额外的半胱氨酸，
其中，任选地，SEQ ID NO：176还包含突变G141S和/或突变P174L。
在一个实施方案中，Xaa198是G，R，K或Y。
在一个实施方案中，Xaa199是P或是缺失的。
在一个实施方案中，SEQ ID NO：171，175或176包含以C对如下氨基酸的取代：
-R47和P174；
-Xaa55和Xaa147；
-Xaa55和Xaa149；
-A59和G71；
-H60和S79；
-S76和S79；
-D107和Xaa155；和/或
-G108和L142。
在一个实施方案中，SEQ ID NO：171，175或176包含以C对如下氨基酸的取代：
-Xaa55和Xaa147；
-Xaa55和Xaa149；
-A59和G71；
-S76和S79；
-D107和Xaa155；或
-G108和L142。
在一个实施方案中，SEQ ID NO：171，175或176包含以C对Xaa55和Xaa147的取代。在另一个实施方案中，SEQ ID NO：171，175或176包含以C对Xaa55和Xaa149的取代。
在一个实施方案中，
-Xaa55是C，Xaa147是C，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是缺失的；
-Xaa55是C，Xaa147是C，Xaa149是N，Xaa155是D，Xaa198是Y，且Xaa199是P；
-Xaa55是C，Xaa147是P，Xaa149是C，Xaa155是D，Xaa198是G，且Xaa199是缺失的；
-Xaa55是C，Xaa147是P，Xaa149是C，Xaa155是D，Xaa198是Y，且Xaa199是P；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是缺失的；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是R，且Xaa199是P；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是K，且Xaa199是P；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是Y，且Xaa199是P；
-Xaa55是C，Xaa147是C，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是P；
-Xaa55是C，Xaa147是P，Xaa149是C，Xaa155是D，Xaa198是G，且Xaa199是P；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是缺失的，其中SEQ ID NO：171，175或176包含以C对A59和G71的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是Y，且Xaa199是P，其中SEQID NO：171，175或176包含以C对A59和G71的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是缺失的，其中SEQ ID NO：171，175或176包含以C对S76和S79的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是Y，且Xaa199是P，其中SEQID NO：171，175或176包含以C对S76和S79的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是缺失的，其中SEQ ID NO：171，175或176包含以C对G108和L142的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是Y，且Xaa199是P，其中SEQID NO：171，175或176包含以C对G108和L142的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是C，Xaa198是G，且Xaa199是缺失的，其中SEQ ID NO：171，175或176包含以C对D107的取代；或
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是C，Xaa198是Y，且Xaa199是P，其中SEQID NO：171，175或176包含以C对D107的取代。
在一个实施方案中，变体在其N末端进一步包含至少一个额外的氨基酸。
在一个实施方案中，所述至少一个额外的氨基酸选自由下列组成的组：G、A、N和C。
在第二方面，本发明涉及人FGF21变体，其包含SEQ ID NO：1-3，172，173和174之一的氨基酸序列或由其组成，其中所述氨基酸序列包含如下突变的至少一个：
Q55G，K150H，P152A，P152L，P152缺失，H153Q，H153Y，H153K，H153缺失，R154Q，R154K，D155L，D155K，D155Y，D155P，D155E，D155N，P156A，P156K，A157V，A157G，A157S，P158H，R159H，R159K，R159Q，A162Y，R163H，Q184缺失，P185缺失，P186缺失，D187缺失，V188缺失，G189缺失，S190缺失，S191缺失，D192缺失，P193缺失，L194缺失，S195缺失，M196缺失，M196P，M196V，V197E，V197D，V197缺失，V197G，V197M，G198E，G198D，G198R，G198K，G198Y，G198P，G198V，G198缺失，P199S，P199Q，P199缺失，P199R，P199T，P199G，P199F，P199L，P199D，P199M，S200Q，S200M，S200P，S200N，S200H，S200缺失，Q201P，Q201S，Q201缺失，G202T，G202缺失，R203E，R203H，R203缺失，S204缺失，P205缺失，S206缺失，Y207缺失，P147-R159由GSGS(SEQ ID NO：161)，P147-R159由GGSGGS(SEQ ID NO：162)，N149-D155由GSGS(SEQ ID NO：161)，N149-D155由GSHSG(SEQ ID NO：163)取代，N149-D155由ATTS(SEQ IDNO：164)取代，N149-R159由GSGS(SEQ ID NO：161)取代，N149-R159由GGSGGS(SEQ ID NO：162)取代，N149-A162由GSHSG(SEQ ID NO：163)取代，N149-A162由GSHSGS(SEQ ID NO：165)取代，N149-R163由GSGS(SEQ ID NO：161)取代，R159-R163由GA，GY，HH，GE或HE取代，V197-R203由GHRSHLQTVF(SEQ ID NO：166)取代，V197-R203由GLNSMV(SEQ ID NO：167)取代，GGGGS(SEQ ID NO：168)插入G198和P199之间，PLSMVGPSQGRSPSYAS(SEQ ID NO：169)或PLSMVGSQGRSPSYAS(SEQ ID NO：170)添加于S209之后；
和/或
其中所述氨基酸序列包含以C对2、4、6或8个如下氨基酸的取代：
R47，L49，T51，A54，Q55，Q56，A59，H60，E62，I63，G67，V69，G71，A72，A73，S76，P77，E78，S79，L80，L81，Q82，L83，I91，L94，G95，V96，K97，T98，R100，L102，Q104，D107，G108，L110，G112，L114，A120，R124，D130，Y132，Q136，S137，A139，H140，L142，P143，H145，L146，P147，G148，N149，D155，P161，A162，L165，L167，L170，P174，S195，M196，S200，R203；
其中所述氨基酸的编号根据SEQ ID NO：1。
在一个实施方案中，所述氨基酸序列包含如下突变的至少一项：
K150H，P152A，P152L，P152缺失，H153Q，H153Y，H153K，H153缺失，R154Q，R154K，D155L，D155K，D155Y，D155P，D155E，D155N，P156A，P156K，A157V，A157G，A157S，P158H，R159H，R159K，R159Q，A162Y，R163H，M196缺失，M196P，M196V，V197E，V197D，V197缺失，V197G，V197M，G198E，G198D，G198R，G198K，G198Y，G198P，G198V，G198缺失，P199S，P199Q，P199缺失，P199R，P199T，P199G，P199F，P199L，P199D，P199M，S200Q，S200M，S200P，S200N，S200H，S200缺失，Q201P，Q201S，Q201缺失，G202T，G202缺失，R203E，R203H，R203缺失；
和/或
所述氨基酸序列包含以C对如下氨基酸的2、4、6或8个的取代：
R47，L49，T51，A54，Q55，Q56，A59，H60，E62，I63，G67，V69，G71，A72，A73，S76，P77，E78，S79，L80，L81，Q82，L83，I91，L94，G95，V96，K97，T98，R100，L102，Q104，D107，G108，L110，G112，L114，A120，R124，D130，Y132，Q136，S137，A139，H140，L142，P143，H145，L146，P147，G148，N149，D155，P161，A162，L165，L167，L170，P174，S195，M196，S200，R203。
在一个实施方案中，所述氨基酸序列包含如下突变的至少一项：
M196缺失，M196P，M196V，V197E，V197D，V197缺失，V197G，V197M，G198E，G198D，G198R，G198K，G198Y，G198P，G198V，G198缺失，P199S，P199Q，P199缺失，P199R，P199T，P199G，P199F，P199L，P199D，P199M，S200Q，S200M，S200P，S200N，S200H，S200缺失，Q201P，Q201S，Q201缺失，G202T，G202缺失，R203E，R203H，R203缺失；
和/或
所述氨基酸序列包含以C对如下氨基酸的2、4、6或8个的取代：
R47，L49，T51，A54，Q55，Q56，A59，H60，E62，I63，G67，V69，G71，A72，A73，S76，P77，E78，S79，L80，L81，Q82，L83，I91，L94，G95，V96，K97，T98，R100，L102，Q104，D107，G108，L110，G112，L114，A120，R124，D130，Y132，Q136，S137，A139，H140，L142，P143，H145，L146，P147，G148，N149，D155，P161，A162，L165，L167，L170，P174。
在一个实施方案中，所述氨基酸序列包含以C对如下氨基酸的取代：
-R47和P174；
-Q55和P147；
-Q55和N149；
-A59和G71；
-H60和S79：
-S76和S79；
-D107和D155；和/或
-G108和L142。
在一个实施方案中，所述氨基酸序列包含选自由下列组成的组的突变：G198R，G198K，G198Y和P199缺失；
和/或
所述氨基酸序列包含以C对如下氨基酸的取代：
-Q55和P147；
-Q55和N149；
-A59和G71；
-S76和S79；
-D107和D155；或
-G108和L142。
在一个实施方案中，所述氨基酸序列包含以C对Q55和P147的取代。
在一个实施方案中，所述氨基酸序列包含以C对Q55和N149的取代。
在一个实施方案中，所述氨基酸序列在SEQ ID NO：1-3，172，173和174之一的另一位点包含至少一个进一步的突变(例如1，2，3，4或5个进一步突变)。
在一个实施方案中，所述氨基酸序列与SEQ ID NO：1-3，172，173和174之一具有至少90％或至少91％或至少92％或至少93％或至少94％或至少95％或至少96％或至少97％或至少98％序列同一性。
在第三方面，本发明涉及人FGF21变体，其包含如下或由如下组成：SEQ ID NO：8，9，10，11，12，13，14，15，16，17，18，19，20，21，22，23，24，25，26，27，28，29，30，31，32，33，34，35，36，37，38，39，40，41，42，43，44，45，46，47，48，49，50，51，52，53，54，55，56，57，58，59，60，61，62，63，64，65，66，67，68，69，70，71，72，73，74，75，76，77，78，79，80，81，82，83，84，85，86，87，88，89，90，91，92，93，94，95，96，97，98，99，100，101，102，103，104，105，106，107，108，109，110，111，112，113，114，115，116，117，118，119，120，121，122，123，124，125，126，127，128，129，130，131，132，133，134，135，136，137，138，139，140，141，142，143，144，145，146，147，148，149，150，151，152，153，154，155，156，157，158，159，160，177，178，179，180，181，182，183，184，185，186，187，188，189，190，191，192，193，194，195，196，197，198，199，200，201，202，203，204，205，206，207，208，209，210，211，212，213，214，215，216，217，218，219，220，221，222，223，224，225，226，227，228，229，230，231，232，233，234，235，236，237，238，239，240，241，242，243，244，245，246，247，248，249，250，251，252，253，254，255，256，257，258，259，260，261，262，263，264，265，266，267，268，269，270，271，272，273，274，275，276，277，278，279，280，281，282，283，284，285，286，287，288，289，290，291，292，293，294，295，296，297，298，299，300，301，302，303，304，305，306，307，308，309，310，311，312，313，314，315，316，317，318，319，320，321，322，323，324，325，326，327和328之一的氨基酸序列，其中，任选地，所述变体进一步包含突变G141S和/或突变P174L，其中氨基酸的编号依据SEQID NO：1。
在一个实施方案中，根据第一、第二和第三方面的变体在人和/或鼠血浆中与成熟人野生型FGF21(SEQ ID NO：2)相比具有增加的蛋白水解稳定性。
在一个实施方案中，变体与成熟人野生型FGF21(SEQ ID NO：2)相比具有增加的热稳定性。
在一个实施方案中，变体诱导促分裂原活化蛋白激酶(MAPK)ERK1/2的磷酸化。
在一个实施方案中，变体以100nmol/L或更低、或90nmol/L或更低、或80nmol/L或更低、或70nmol/L或更低、或60nmol/L或更低、或50nmol/L或更低、或40nmol/L或更低、或30nmol/L或更低、或20nmol/L或更低、或15nmol/L或更低、或12nmol/L或更低、或11nmol/L或更低、或10nmol/L或更低、或9nmol/L或更低、或8nmol/L或更低、或7nmol/L或更低、或6nmol/L或更低、或5nmol/L或更低、或4nmol/L或更低、或3nmol/L或更低、或2nmol/L或更低的EC50诱导促分裂原活化蛋白激酶(MAPK)ERK1/2的磷酸化，例如，所述EC50如In-CellWestern(ICW)测定法中，如在哺乳动物细胞培养中确定的。
在一个实施方案中，变体进一步包含至少一个允许对所述变体进行检测和/或分离的标记或标签。
在一个实施方案中，变体融合或缀合至半衰期延长模块。
在一个实施方案中，所述半衰期延长模块选自由下列组成的组：聚合物(例如聚乙二醇(PEG)，羟乙基淀粉(HES)，透明质酸，聚唾液酸)，非结构化(多)肽链(例如PAS，XTEN)，弹性蛋白样多肽(ELP)，血清蛋白(例如白蛋白)，血清蛋白结合分子(例如白蛋白结合域(ABD)，白蛋白结合脂肪酸)，抗体，免疫球蛋白，免疫球蛋白的Fc区/域和免疫球蛋白结合域。
在另一个方面，本发明涉及融合分子，其包含如上文定义的人FGF21变体及至少一种其他活性药物成分。
在另一个方面，本发明涉及核酸分子，其编码如上文定义的人FGF21变体或如上文定义的融合分子。
在一个实施方案中，所述核酸分子包含于载体中或整合至基因组中。
在另一个方面，本发明涉及宿主细胞，其含有如上文定义的核酸分子。
在另一个方面，本发明涉及生产如上文定义的人FGF21变体或如上文定义的融合分子的方法，其包括培养如上文定义的宿主细胞并从培养基分离所述变体或融合分子。
在另一个方面，本发明涉及药物组合物，其包含如上文定义的人FGF21变体或如上文定义的融合分子或如上文定义的核酸分子或如上文定义的宿主细胞，以及医药上可接受之载剂和/或赋形剂。
在一个实施方案中，所述药物组合物进一步包含至少一种其他活性药物成分。
在另一个方面，本发明涉及包含如上文定义的人FGF21变体与至少一种其他活性药物成分的组合。
在一个实施方案中，如上文定义的融合分子或药物组合物或组合中包含的至少一种其他活性药物成分选自由下列组成的组：胰岛素和胰岛素衍生物，GLP-1，GLP-1类似物和GLP-1受体激动剂，聚合物结合的GLP-1和GLP-1类似物，双重GLP-1/GIP激动剂，双重GLP-1/胰高血糖素受体激动剂，PYY3-36或其类似物，胰多肽或其类似物，胰高血糖素受体激动剂或拮抗剂，GIP受体激动剂或拮抗剂，胃饥饿素(ghrelin)拮抗剂或反向激动剂，Xenin及其类似物，(特异性)结合上述受体(例如，GLP-1受体，GIP受体，胰高血糖素受体)之一的肽，DDP-IV抑制剂，SGLT-2抑制剂，双重SGLT-2/SGLT-1抑制剂，双胍类，噻唑烷二酮类，PPAR激动剂，PPAR调节剂，磺酰脲类，氯茴苯酸类，α-葡萄糖苷酶抑制剂，胰淀素及胰淀素类似物，GPR119激动剂，GPR40激动剂，GPR120激动剂，GPR142激动剂，TGR5激动剂，AMPK刺激剂，AMPK激活剂，11-β-HSD抑制剂，葡萄糖激酶激活剂，DGAT抑制剂，蛋白酪氨酸磷酸酶1抑制剂，葡萄糖-6-磷酸酶抑制剂，果糖-1，6-二磷酸酶抑制剂，糖原磷酸化酶抑制剂，磷酸烯醇丙酮酸羧激酶抑制剂，糖原合酶激酶抑制剂，丙酮酸脱氢酶激酶抑制剂，CCR-2拮抗剂，葡萄糖转运蛋白-4调节剂，生长抑素受体3激动剂，HMG-CoA-还原酶抑制剂，贝特类，烟酸及其衍生物，烟酸受体1激动剂，，ACAT抑制剂，胆固醇吸收抑制剂，胆汁酸结合物质，IBAT抑制剂，MTP抑制剂，PCSK9调节剂，LDL受体上调调节剂(肝选择性甲状腺激素受体β激动剂)，HDL升高化合物，脂质代谢调节剂，PLA2抑制剂，ApoA-I增强剂，胆固醇合成抑制剂，ω-3脂肪酸及其衍生物，用于治疗肥胖症的活性物质，CB1受体拮抗剂，MCH-1拮抗剂，MC4受体激动剂和部分激动剂，NPY5或NPY2拮抗剂，NPY4激动剂，β3肾上腺素能受体激动剂，瘦蛋白或瘦蛋白模拟物，5HT2c受体激动剂，脂肪酶抑制剂，血管生成抑制剂，H3拮抗剂，AgRP抑制剂，三重单胺摄取抑制剂，MetAP2抑制剂，针对成纤维细胞生长因子受体4或阻抑素(prohibitin)靶向肽-1生产的反义寡核苷酸，用于影响高血压、慢性心力衰竭或动脉粥样硬化的药物，血管紧张素II受体拮抗剂，双重血管紧张素受体阻断剂(ARB)，血管紧张素转化酶(ACE)抑制剂，血管紧张素转化酶2(ACE-2)活化剂，，肾素抑制剂，肾素原抑制剂，内皮素转化酶(ECE)抑制剂，内皮素受体阻断剂，内皮素拮抗剂，利尿剂，醛固酮拮抗剂，醛固酮合酶抑制剂，α-阻断剂，α-2肾上腺素能受体拮抗剂，β-阻断剂，混合的α/β-阻断剂，钙拮抗剂/钙通道阻断剂(CBB)，双重盐皮质激素/CCB，中枢作用的抗高压药，中性内肽酶抑制剂，氨基肽酶-A抑制剂，血管肽(vasopeptide)抑制剂，双重血管肽抑制剂，脑啡肽酶(neprilysin)-ACE抑制剂，脑啡肽酶-ECE抑制剂，双重作用血管紧张素(AT)受体-脑啡肽酶抑制剂，双重AT1/内皮素-1(ETA)拮抗剂，晚期糖基化终末产物破坏剂，重组肾酶，血压疫苗，抗RAAS疫苗，AT1-或AT2-疫苗，具有抗高血压响应的遗传多态性调节剂和血小板聚集抑制剂。
在一个实施方案中，所述至少一种其他活性药物成分是胰高血糖素样肽-1(GLP-1)受体激动剂。
在另一个方面，本发明涉及试剂盒，其包含如上文定义的人FGF21变体或如上文定义的融合分子或如上文定义的核酸分子或如上文定义的宿主细胞或如上文定义的药物组合物。
在另一个方面，本发明涉及如上文定义的人FGF21变体或如上文定义的融合分子或如上文定义的核酸分子或如上文定义的宿主细胞或如上文定义的药物组合物，其用作药物。
在另一个方面，本发明涉及如上文定义的人FGF21变体或如上文定义的融合分子或如上文定义的核酸分子或如上文定义的宿主细胞或如上文定义的药物组合物，其用于治疗选自由下列组成的组的疾病或病症：肥胖症，超重，代谢综合征，糖尿病，高血糖症，血脂异常，非酒精性脂肪性肝炎(NASH)和动脉粥样硬化。
在另一个方面，本发明涉及如上文定义的人FGF21变体或如上文定义的融合分子或如上文定义的核酸分子或如上文定义的宿主细胞或如上文定义的药物组合物在制造用于治疗选自由下列组成的组的疾病或病症的药物中的用途：肥胖症，超重，代谢综合征，糖尿病，高血糖症，血脂异常，非酒精性脂肪性肝炎(NASH)和动脉粥样硬化。
在另一个方面，本发明涉及用于治疗选自由下列组成的组的疾病或病症的方法：肥胖症，超重，代谢综合征，糖尿病，高血糖症，血脂异常，非酒精性脂肪性肝炎(NASH)和动脉粥样硬化，该方法包括将如上文定义的人FGF21变体或如上文定义的融合分子或如上文定义的核酸分子或如上文定义的宿主细胞或如上文定义的药物组合物施用于对其有需要的受试者。
在一个实施方案中，所述疾病或病症是糖尿病。
附图说明
图1显示了SEQ ID NO：3的FGF21及其代谢物的质谱和解卷积(deconvolution)数据。(A)SEQ ID NO：3的氨基酸[1-182]的序列的质谱和解卷积；(B)人血浆中(也见于大鼠和鼠类血浆中)SEQ ID NO：3的氨基酸[1-172]的序列的质谱和解卷积；RT：33.6分钟；以C标记的多电荷质量(multicharged masses)解卷积至18,429Da；(C)人血浆中(也见于大鼠血浆中)SEQ ID NO：3的氨基酸[132-182]的序列的质谱和解卷积；RT：30.4分钟；以F标记的多电荷质量解卷积至5，113.67Da；(D)人血浆中SEQ ID NO：3的氨基酸[129-182]，[125-177]，[126-178]或[124-176]的序列的质谱和解卷积；RT：30.8分钟；以E标记的多电荷质量解卷积至5，379Da；(E)人血浆中SEQ ID NO：3的氨基酸[136-182]的推测(proposed)序列的质谱和解卷积；RT：31.3分钟；以B标记的多电荷质量解卷积至4,732.41Da；(F)鼠血浆中SEQ IDNO：3的氨基酸[53-129]或[21-98]的推测序列的质谱和解卷积；RT：32.5；以A标记的多电荷质量解卷积至8,564.67Da。
图2显示了人血浆中成熟人FGF21(SEQ ID NO：2)和FGF21变体的稳定性分析。将所述蛋白在血浆中于37℃温育24小时，并用识别完整成熟FGF21的ELISA测量剩余的FGF21量。温育开始时的量设为100％。数据为均值+SD。对应的SEQ ID NO显示于表3。
图3显示了在用成熟人FGF21(SEQ ID NO：2)或多种FGF21变体刺激后过表达人FGFR1c和β-klotho的CHO细胞中ERK1/2-磷酸化的剂量-响应曲线，通过In-Cell Western(A至W)测量。对应的SEQ ID NO显示于表4。
图4显示了在用成熟人FGF21(SEQ ID NO：2)或多种FGF21变体刺激后原发人皮下脂肪细胞中ERK1/2-磷酸化的剂量-响应曲线，通过In-Cell Westem (A和B)测量。对应的SEQ ID NO显示于表5。
图5显示了人FGF21(SEQ ID NO：3)和FGF21变体的热稳定性分析，其通过差示扫描荧光分析(DSF)来加以分析。显示了来自针对人FGF21(SEQ ID NO：3)和选出的FGF21变体的ThermoFluorTM实验的示例性的熔解曲线。将标准化于25℃的相对荧光单位针对温度(℃)绘图。对应的SEQ ID NO显示于表6。
图6显示了人FGF21的计算和实验模型的结构细节。(A)以基于FGF19和FGF23的不同实验同源结构的主链结构表示的计算模型叠加，白色：用BioLuminate基于FGF19(PDB码2P23)的结构生成的模型；浅灰色：用Swiss-Model服务器基于FGF19(PDB码1PWA)的结构生成的模型；深灰色：用Swiss-Model服务器基于FGF19(PDB码2P23)的结构生成的模型；黑色：用Swiss-Model服务器基于FGF23(PDB码2P39)的结构生成的模型。尤其是残基150-158(环绕的)周围的环区与这些模型中的显著不同，这是由于基本的(underlying)实验数据的结构差异所致。(B)FGF21 Q55C、N149C(SEQ ID NO：158)的X射线晶体结构中存在的分子A(浅灰色)和分子B(深灰色)的叠加，以漫画显示。半胱氨酸149和55之间工程化的二硫键将构象稳定化，由此增加热稳定性，其突出显示。(C)分子B中围绕工程化的C55-C149二硫键的2mFo-dFc电子密度。等高水平(contour level)为1σ。在C149和P156之间，电子密度是不连续的，提示了显著的紊乱。
发明详述
虽然本发明在下文进行了详细描述，但应当理解本发明并不限于此处描述的具体的方法、方案和试剂，而是可以变化。还应当理解的是，本文所用的术语仅出于描述具体实施方案的目的，而非意欲限制本发明的范围，本发明的范围仅由所附的权利要求书限定。除另有定义外，本文所用的所用技术和科学术语具有如本领域普通技术人员所理解的相同的含义。
在下文中，将描述本发明的一些要素。这些要素可以用具体实施方案列出，然而应当理解，它们可以以任何方式和任何数量组合以产生另外的实施方案。各种描述的实例和实施方案不应被解释为将本发明仅限于明确描述的实施方案。这种描述应当理解为支持并涵盖将明确描述的实施方案与任何数量的所公开的要素组合在一起的实施方案。此外，本申请中所有描述的要素的任何排列和组合应当视为通过本申请的描述而公开，除上下文另有说明外。
本文所用的术语一般定义如“A multilingual glossary of biotechnologicalterms：(IUPAC推荐)”，H.G.W.Leuenberger，B.Nagel，and H.Eds.，HelveticaChimica Acta，CH-4010Basel，Switzerland，(1995)中所述。
除另有说明外，本发明的实践采用常规的化学、生物化学、细胞生物学、免疫学方法、和重组DNA技术，其解释于本领域的文献中(Sambrook，J等(2001)Molecular Cloning：ALaboratory Manual，第三版，Cold Spring Harbor Laboratory Press，Cold SpringHarbor，NY)。
在本说明书及所附的权利要求书中，除上下文另有要求外，词语“包含”，及其变体“包括”和“含有”，理解为包含所述的成员、整数或步骤或成员、整数或步骤的组，但不排除任何其他成员、整数或步骤或成员、整数或步骤的组，尽管在一些实施方案中，此类其他成员、整数或步骤或成员、整数或步骤的组可以被排除，即所述主题由所述的成员、整数或步骤或步骤或成员、整数或步骤的组组成。术语“一个”和“一种”和“所述”及描述本发明的上下文中所使用的类似指代(尤其在权利要求中)应理解为涵盖单数和复数，除本文另有说明或与上下文明显矛盾外。本文的值的范围的记载仅意在作为分别提及落在该范围中的各个单独的值的简化记法(shorthand method)。除非本文另有说明，否则每个单独的值并入本说明书中，如同其在本文中被单独记载一样。本文所述的所有方法可以以任何合适的顺序进行，除非本文另有说明或上下文明显矛盾。本文提供的任何和所有实例或示例性语言(例如“如”)的使用仅意在更好地说明本发明，而非对所要求保护的本发明的范围构成限制。说明书中的语言不应被解释为指示对本发明的实践必要的任何未要求保护的要素。
在本说明书的整个文本中引用了一些文件。无论是上文还是下文，本文引用的每篇文献(包括所有专利，专利申请，科学出版物，制造商的说明书，说明书等)都通过提述以其整体并入本文。本文中的任何内容不应被解释为承认本发明由于在先发明而没有早于这样的公开。
如本文所用的，术语“人成纤维细胞生长因子21(FGF21)”是指具有SEQ ID NO：1的氨基酸序列的人野生型人野生型FGF21(也称为“全长人野生型FGF21”)或其天然存在的变体。术语“成熟人野生型FGF21”意指缺乏天然信号序列(也称为信号肽，即SEQ ID NO：1的氨基酸1至28(M1至A28))的人野生型FGF21，且由SEQ ID NO：2表示。除另有说明外，本文公开的FGF21(多)肽中的具体氨基酸残基的编号依据SEQ ID NO：1。
术语“天然存在的”在与生物材料(如核酸分子、(多)肽、宿主细胞等等)相关使用时，意指在自然界中发现而非人工操作的材料。
在向公众开放的数据库中，例如通用蛋白质资源(Universal Protein Resource)(UniProt，在线数据库可从uniprot.org获得)描述了人FGF21的一些天然存在的变体。在UniProtKB登录号Q9NSA1下列出了两个人FGF21：FGF21 G141→S(VAR_055375，SEQ ID NO：172)和FGF21L174→p(VAR_049064)。根据数据库，位置174处脯氨酸的出现比亮氨酸更常见，如ExAC浏览器(P＝73％对L＝27％；The Exome Aggregation Consortium；在线可获取自exac.broadinstitute.org/variant/19-49261368-T-C)和AMP(AcceleratingMedicines Partnership；在线可获取自attype2diabetesgenetics.org/variantInfo/variantInfo/rs739320)。因此，具有P174的人FGF21在本文中被称为人野生型FGF21(参见SEQ ID NO：1)，而具有L174的人FGF21包括与本文中作为天然存在的变体(参见SEQ ID NO：173)。位置174处的突变根据AMP不影响2型糖尿病的发生。此外，L174突变为P对通过In-Cell Western(在实施例3中描述)评估的FGF21的体外活性没有影响。在位置174处具有脯氨酸的G-FGF21(SEQ ID NO：3)的EC 50为0.24hmol/L，相比之下在位置174处具有亮氨酸(SEQ ID NO：174)的G-FGF21的EC 50为0.21nmol/L。FGF21变体G141→S(VAR_055375，SEQID NO：172)是非常少见的，其等位基因频率低于0.1％(在线可获取自exac.broadinstitute.org/variant/19-49261268-G-A)且未在体外进行进一步表征。
根据本发明的“人成纤维细胞生长因子21(FGF21)的变体”(也称为“FGF21变体”或“FGF21类似物”)是具有形式上可以衍生自人野生型FGF21或其天然存在的变体的结构的多肽，其通过缺失和/或取代天然存在的人FGF21中的至少一个氨基酸残基(例如SEQ ID NO：1或2)和/或添加至少一个氨基酸残基而形成。
在一个实施方案中，人FGF21的变体是人FGF21的生物活性变体。术语“人FGF21的生物活性变体”是指具有人野生型FGF21多肽活性的任何变体，例如降低血糖，胰岛素，甘油三酯或胆固醇，降低体重和/或改善葡萄糖耐量、能量消耗或胰岛素敏感度的能力。相对于人野生型FGF21多肽具有稍微降低的FGF21活性水平的变体仍然可以视为人FGF21的生物活性变体。
在一些实施方案中，人野生型FGF21氨基酸序列(例如，SEQ ID NO：1或2的氨基酸序列)的具体氨基酸可以被另一氨基酸取代。本文所用的术语“另一氨基酸”通常涉及与人野生型FGF21(例如SEQ ID NO：NO：1或2)相比有助于变体的增加的稳定性(例如蛋白水解稳定性和/或热稳定性)的氨基酸。这可以例如通过防止在取代的氨基酸处或附近的蛋白酶裂解，或通过一个或多个额外二硫键的形成来实现。
本文所用的术语“氨基酸”或“氨基酸残基”是指天然存在的氨基酸，非天然氨基酸，氨基酸类似物和氨基酸模拟物，其以与天然存在的氨基酸类似的方式起作用，均呈现其D和L立体异构体，如果它们的结构允许这样的立体异构形式的话。氨基酸在本文中以其名称(即其众所周知的三字母符号)来表示，或通过IUPAC-IUBBiochemical NomenclatureCommission推荐的单字母符号来表示。
当与氨基酸结合使用时，术语“天然存在的”是指20种常规氨基酸(即丙氨酸(A)，半胱氨酸(C)，天冬氨酸(D)，谷氨酸(E)，苯丙氨酸(F)，甘氨酸(G)，组氨酸(H)，异亮氨酸(I)，赖氨酸(K)，亮氨酸(L)，甲硫氨酸(M)，天冬酰胺(N)，脯氨酸(P)，谷氨酰胺(Q)，精氨酸(R)，丝氨酸(S)，苏氨酸(T)，缬氨酸(V)，色氨酸(W)和酪氨酸(Y))，以及硒代半胱氨酸，吡咯赖氨酸(PYL)和吡咯啉羧基赖氨酸(PCL)。
本文所用的术语“非天然氨基酸”意指不是天然编码的或在任何生物体的遗传密码中发现的氨基酸。它们可以是例如纯合成的化合物。非天然氨基酸的实例包括但不限于，羟基脯氨酸，γ-羧基谷氨酸，O-磷酸丝氨酸，氮杂环丁烷羧酸，2-氨基己二酸，3-氨基己二酸，β-丙氨酸，氨基丙酸，2-氨基丁酸，4-氨基丁酸，6-氨基己酸，2-氨基庚酸，2-氨基异丁酸，3-氨基异丁酸，2-氨基庚二酸，叔丁基甘氨酸，2，4-二氨基异丁酸，锁链素(desmosine)，2，2′-二氨基庚二酸，2，3-二氨基丙酸，N-乙基甘氨酸，N-甲基甘氨酸，N-乙基天冬酰胺，高脯氨酸，羟基赖氨酸，别羟基赖氨酸(allo-hydroxylysine)，3-羟基脯氨酸，4-羟基脯氨酸，异锁链素(isodesmosine)，别异亮氨酸，N-甲基丙氨酸，N-甲基甘氨酸，N-甲基异亮氨酸，N-甲基戊基甘氨酸，N-甲基缬氨酸，萘基丙氨酸(naphthalanine)，正缬氨酸，正亮氨酸，鸟氨酸，D-鸟氨酸，D-精氨酸，对氨基苯丙氨酸，戊基甘氨酸，呱啶酸(pipecolic acid)和硫代脯氨酸。
本文所用的术语“氨基酸类似物”是指具有与天然存在的氨基酸相同的基本化学结构的化合物。氨基酸类似物包括被化学阻断(可逆地或不可逆地)的天然和非天然氨基酸，或其C-末端羧基，其N-末端氨基和/或其侧链官能团被化学修饰。这种类似物包括但不限于甲硫氨酸亚砜，甲硫氨酸砜，S-(羧甲基)-半胱氨酸，S-(羧甲基)-半胱氨酸亚砜，S-(羧甲基)-半胱氨酸砜，天冬氨酸-(β甲酯)，N-乙基甘氨酸，丙氨酸羧酰胺，高丝氨酸，正亮氨酸和甲硫氨酸甲基锍(methioninemethyl sulfonium)。
本文所用的术语“氨基酸模拟物”是指结构不同于氨基酸的一般化学结构的化学化合物，但其以与天然存在的氨基酸类似的方式起作用。
本文所用的术语“肽”是指任何长度的氨基酸的聚合形式，例如包含两个或更多个，或3个或更多个，或4个或更多个，或6个或更多个，或8个或更多个，或9个或更多个，或10个或更多个，或13个或更多个，或16个或更多个，或21个或更多个氨基酸，其通过肽键共价连接。术语“多肽”是指大的肽，例如具有超过100个氨基酸残基的肽。术语“多肽”和“蛋白质”在本文中可互换使用。
在一些实施方案中，所述变体在其N末端包含至少一个额外的氨基酸。在一个实施方案中，所述至少一个额外的氨基酸选自除脯氨酸外的天然存在的氨基酸，非天然氨基酸，氨基酸类似物和氨基酸模拟物。在一个实施方案中，所述至少一种额外的氨基酸选自G、A、N和C。在一个具体的实施方案中，所述至少一个额外的氨基酸是G。
在一些实施方案中，本发明的变体的氨基酸序列与SEQ ID NO：1-3、172、173和174(例如与成熟人野生型FGF21(SEQ ID NO：2))具有至少90％或至少91％或至少92％或至少93％或至少94％或至少95％或至少96％或至少97％或至少98％序列同一性。
两个氨基酸序列之间的“序列同一性”表示序列之间相同的氨基酸的百分比。除了手动以外，可以通过Smith和Waterman，1981，Ads App.Math.2，482的局部同源性算法、通过Neddleman和Wunsch，1970，J.Mol.Biol.48，443的局部同源性算法、通过Pearson和Lipman，1988，Proc.Natl Acad.Sci.USA 85，2444的相似性搜索方法、或通过使用这些算法的计算机程序(Wisconsin Genetics软件包中的GAP，BESTFIT，FASTA，BLAST P，BLAST N和TFASTA，Genetics Computer Group，575Science Drive，Madison，Wis.)来产生用于比较的最佳序列比对。
与成熟人野生型FGF21(SEQ ID NO：2)相比，本发明表征的FGF21变体通常在人和/或鼠血浆中具有增加的蛋白水解稳定性。
术语“蛋白水解稳定性”是指(多)肽耐受蛋白酶催化的蛋白水解的能力。这种蛋白酶的实例是丝氨酸蛋白酶，苏氨酸蛋白酶，半胱氨酸蛋白酶，天冬氨酸蛋白酶和/或谷氨酸蛋白酶。丝氨酸蛋白酶的实例是胰蛋白酶，糜蛋白酶，纤溶酶，凝血酶，粒酶和/或激肽释放酶(kallikreine)。半胱氨酸蛋白酶的实例是组织蛋白酶K，胱天蛋白酶和/或钙蛋白酶。根据EC编号，此类蛋白酶例如根据酶活性分类为丝氨酸羧肽酶(EC 3.4.16)，丝氨酸内肽酶(EC 3.4.21)，苏氨酸内肽酶(EC 3.4.25)，半胱氨酸羧肽酶(EC 3.4.18)，半胱氨酸内肽酶(EC 3.4.22)或天冬氨酸内肽酶(EC 3.4.23)。术语“蛋白水解稳定性”可以指体外或体内蛋白水解稳定性。测量给定(多)肽的蛋白水解稳定性的手段和方法是本领域技术人员已知的。在一个实施方案中，蛋白水解稳定性用夹心免疫测定法测定，例如基本上如实施例2所述。
本文所用的术语“蛋白酶抗性肽接头”是指对蛋白酶(例如上述蛋白酶)催化的蛋白水解具有低易感性或耐受蛋白水解的肽接头。这种接头可以具有1至20或1至15或1至12或1至10个氨基酸的长度。在一些实施方案中，接头可以由少至两个氨基酸(例如，GA，GY，HH，GE或HE)组成。在一个具体实施方案中，蛋白酶抗性肽接头是SEQ ID NO：161，SEQ IDNO：162，SEQ ID NO：163，SEQ ID NO：164，SEQ ID NO：165，SEQ ID NO：166，SEQ ID NO：167，或SEQ ID NO：168的肽接头。
与成熟人野生型FGF21(SEQ ID NO：2)相比，本发明表征的FGF21变体通常具有增加的热稳定性。
术语“热稳定性”是指(多)肽在较高温度(例如高于40℃)抵抗其化学或物理结构的不可逆变化的能力。在具体实施方案中，增加的热稳定性是指变体的增加的解链温度(TM)。测量给定(多)肽的热稳定性的手段和方法是本领域技术人员已知的。在一个实施方案中，在生理缓冲液(如PBS)中测量热稳定性(例如TM)。在一个实施方案中，用热迁移测定法测量热稳定性(例如TM)，例如基本上如实施例5中所述。
在一个实施方案中，变体的熔解温度(TM)与成熟人野生型FGF21(SEQ ID NO：2)相比增加至少2℃、至少4℃、至少6℃、至少8℃、至少10℃、至少12℃、至少14℃、至少16℃、至少18℃或至少20℃。在一个实施方案中，TM与成熟人野生型FGF21(SEQ ID NO：2)相比增加至少18℃，例如18℃至30℃或18℃至25℃。
FGF21变体的生物学功能或活性通常与人野生型FGF21(例如成熟人野生型FGF21(SEQ ID NO：2))在本领域技术人员普遍已知的FGF21活性测定法中进行测量和/或比较。FGF21活性测定法是例如“葡萄糖摄取测定法”，如Kharitonenkov A等(2005)J ClinInvest 115：1627-1635中所述。或者，细胞测定法，例如自动磷酸化测定法，例如In-CellWestern(ICW)测定法，可用于测量FGF21变体激活FGF受体：KLB复合物的FGF21受体或刺激下游细胞内信号转导的功效。
本文所用的术语“In-Cell Western(ICW)测定”是指通常在微孔板(例如96孔或384孔规格)中进行的免疫细胞化学测定，更具体地是定量免疫荧光测定。它结合了Western印迹的特异性与ELISA的可重复性和通量(参见例如Aguilar H.N.等(2010)PLoS ONE 5(4)：e9965)。合适的ICW测定系统可在商业上获得(例如获取自LI-COR Biosciences，USA)。在一个实施方案中，ICW测定基本上如实施例3或4中所述进行。
通常，FGF21变体诱导丝裂原活化蛋白激酶(MAPK)ERK1/2的磷酸化。在一个实施方案中，变体以100nmol/L或更低、或90nmol/L或更低、或80nmol/L或更低、或70nmol/L或更低、或60nmol/L或更低、或50nmol/L或更低、或40nmol/L或更低、或30nmol/L或更低、或20nmol/L或更低、或15nmol/L或更低、或12nmol/L或更低、或11nmol/L或更低、或10nmol/L或更低、或9nmol/L或更低、或8nmol/L或更低、或7nmol/L或更低、或6nmol/L或更低、或5nmol/L或更低、或4nmol/L或更低、或3nmol/L或更低、或2nmol/L或更低的EC50(如In-CellWestern(ICW)测定所测得的)诱导丝裂原活化蛋白激酶(MAPK)ERK1/2的磷酸化。
“允许对变体进行检测和/或分离的标记或标签”意在包括本领域已知的用于这些目的任何标记/标签。示例性标签包括亲和标签，例如几丁质结合蛋白(CBP)，麦芽糖结合蛋白(MBP)，谷胱甘肽-S-转移酶(GST)和His-标签，如富组氨酸序列(例如SEQ ID NO：5或6)或聚(His)(例如His6)；增溶标签，如硫氧还蛋白(TRX)和聚(NANP)；色谱标签，如FLAG标签；表位标签，如V5标签，myc-标签和HA-标签；和荧光或发光标记或标签，例如荧光蛋白(例如GFP，YFP，RFP等)，荧光染料和荧光素酶。
(多)肽标记或标签的氨基酸序列可以引入变体的氨基酸序列内的任何位置处，并且可以例如在编码的蛋白结构内具有环的形状，或者其可以融合于N末端或C末端。在一个实施方案中，标记或标签是N末端融合的。标记或标签可以进一步包含允许从变体中除去标记或标记的切割位点(例如TEV蛋白酶切割位点)。类似地，非肽标记或标签，例如荧光染料，可以在任何合适的位点与变体缀合。
本发明表征的FGF21变体还可以包含用于促进分子分泌的氨基酸序列，例如N-末端分泌信号。在一些实施方案中，分泌信号是允许充分通过分泌途径和/或分泌进入细胞外环境的信号序列。在一些实施方案中，分泌信号序列是可切割的，并从变体除去。在一个实施方案中，信号序列不同于人FGF21的天然信号序列。分泌信号序列适用于其中产生变体的细胞或生物体。在一个实施方案中，分泌信号序列包含SEQ ID NO：4的氨基酸序列或由其组成。
本发明的FGF21变体可以进一步包含用于例如增强对特定细胞类型或组织的选择性的结合域。这可以例如通过提供结合域来实现，所述结合域结合在所述细胞类型或组织的表面上表达的特异性抗原。
FGF21变体也可以与半衰期延长模块融合或缀合。“半衰期”涉及(例如在体内)消除分子的一半的活性、量或数目所需的时间段。此类模块是本领域技术人员已知的并且包括例如聚合物(例如聚乙二醇(PEG)，羟乙基淀粉(HES)，透明质酸，聚唾液酸)，非结构化(多)多肽链，弹性蛋白样多台(ELP)，血清蛋白(例如白蛋白，例如人血清白蛋白(HAS))，血清蛋白结合分子(例如白蛋白结合域(ABD)，白蛋白结合脂肪酸)，抗体，免疫球蛋白，免疫球蛋白的Fc区/域和免疫球蛋白结合域。
本文所用的术语“非结构化(多)肽链”是指缺少固定或有序三维结构且通常是亲水性的(多)肽链。延长其融合的肽和蛋白质(体内)半衰期的非结构化(多)肽链是本领域技术人员已知的，包括例如XTEN(Schellenberger V等(2009)Nat Biotechnol.27(12)：1186-90)和PAS序列(Schlapschy M.等(2013)Protein Eng Des Sel.26(8)：489-501)。
本文所用的术语“与......融合/融合至”具体是指例如通过重组DNA技术的遗传融合。可以将(聚)肽半衰期延长模块的氨基酸序列引入变体的氨基酸序列内任何位置，并且可以例如在编码的蛋白结构内具有环的形状，或者其可以融合于N-末端或C-末端。
本文所用的术语“与......缀合/缀合至”具体是指在(多)肽与另一分子(例如变体和半衰期延长模块)之间产生稳定的共价连接的化学和/或酶缀合。这种缀合可以发生在(多)肽的N-或C-末端或特定侧链，例如在赖氨酸，半胱氨酸，酪氨酸或非天然氨基酸残基处。
术语“融合分子”通常是指通过接合，特别是共价连接两个或更多个不同分子(例如蛋白质和/或肽)而产生的分子，其产生具有来源于每个初始分子的功能特性的单个分子。在蛋白质和/或肽的情况下，融合分子称为“融合蛋白”。融合分子可以通过遗传融合(例如通过重组DNA技术)或通过化学和/或酶缀合产生。两个或更多个不同分子还可以通过合适的接头分子连接，例如肽接头或非肽聚合物，如聚乙二醇(PEG)。肽接头还可以包含特异性蛋白酶切割位点。
根据本发明，除了FGF21变体外，融合分子还包含至少一种其它活性药物成分。
本文使用的术语“活性药物成分”(API)包括任何药学活性的化学或生物化合物及其任何医药上可接受之盐和其任何混合物，其提供一些药理作用并用于治疗或预防病况，例如本文定义的疾病或病症。示例性的医药上可接受之盐包括盐酸，硫酸，硝酸，磷酸，氢溴酸，马来酸(maleric)，苹果酸，抗坏血酸，柠檬酸，酒石酸，扑酸(pamoic)，月桂酸，硬脂酸，棕榈酸，油酸，肉豆蔻酸，月桂基硫酸，萘磺酸(naphthalinesulfonic)，亚油酸，亚麻酸等。如本文所用的，术语“活性药物成分”，“活性剂”，“活性成分”，“活性物质”，“治疗活性化合物”和“药物”是指同义词，即具有相同的含义。
根据本发明，活性药物成分任选地选自：
-Rote Liste 2014中提及的所有药物，例如在Rote Liste 2014，第12章中提及的所有抗糖尿病药或在Rote Liste 2014，第06章中提及的所有减肥药或食欲抑制剂，在RoteListe 2014，第58章中提及的所有降脂药，Rote Liste 2014第17章中提及的所有降压药，Rote Liste中提及的所有肾保护剂，或Rote Liste 2014，第36章中提及的所有利尿剂；
-胰岛素和胰岛素衍生物，例如：甘精胰岛素(例如)，高于100U/mL的浓缩甘精胰岛素，例如270-330U/mL的甘精胰岛素或300U/mL的甘精胰岛素(如EP 2387989中所公开的)，谷赖胰岛素(例如)，地特胰岛素(例如)，赖脯胰岛素(例如)，德谷胰岛素(insulin degludec)(例如IdegLira(NN9068))，门冬胰岛素和门冬胰岛素制剂(例如)，基础胰岛素和类似物(例如LY2605541，LY2963016，NN1436)，聚乙二醇化赖脯胰岛素(例如LY-275585)，长效胰岛素(例如NN1436，Insumera(PE0139)，AB-101，AB-102，Sensulin LLC)，中效胰岛素(例如 N， N)，速效胰岛素和短效胰岛素(例如 R， R，PH20 insulin，NN1218，)，预混合胰岛素，NN1045，胰岛素加PE-0139，ACP-002水凝胶胰岛素，和口服、吸入式、经皮和口腔或舌下胰岛素(例如 tregopil胰岛素，TPM-02胰岛素， 口服胰岛素，ORMD-0801，Oshadi口服胰岛素，NN1953，NN1954，NN1956，)。同样合适的还有通过双功能接头与白蛋白或另一蛋白键合的那些胰岛素衍生物；
-胰高血糖素样肽1(GLP-1)，GLP-1类似物和GLP-1受体激动剂，例如：GLP-1(7-37)，GLP-1(7-36)酰胺，利西拉来(例如)，艾塞那肽(例如毒蜥外泌素-4(exendin-4)，重组毒蜥外泌素-4(rExendin-4)， 艾塞那肽NexP)，艾塞那肽-LAR，利拉鲁肽(例如)，司美鲁肽(semaglutide)，他司鲁泰(taspoglutide)，阿必鲁泰(albiglutide)，度拉糖肽(dulaglutide)，albugon，胃泌酸调节素，geniproside，ACP-003，CJC-1131，CJC-1134-PC，GSK-2374697，PB-1023，TTP-054，langlenatide(HM-11260C)，CM-3，GLP-1Eligen，AB-201，ORMD-0901，NN9924，NN9926，NN9927，Nodexen，Viador-GLP-1，CVX-096，ZYOG-1，ZYD-1，ZP-3022，CAM-2036，DA-3091，DA-15864，ARI-2651，ARI-2255，艾塞那肽-XTEN(VRS-859)，艾塞那肽-XTEN+胰高血糖素-XTEN(VRS-859+AMX-808)和聚合物结合的GLP-1和GLP-1类似物；
-双重GLP-1/GIP激动剂(例如RG-7697(MAR-701)，MAR-709，BHM081，BHM089，BHM098)；双重GLP-1/胰高血糖素受体激动剂(例如BHM-034，OAP-189(PF-05212389，TKS-1225)，TT-401/402，ZP2929，LAPS-HMOXM25，MOD-6030)；
-双重GLP-1/胃泌素激动剂(例如ZP-3022)；
-胃肠肽如肽YY 3-36(PYY3-36)或其类似物和胰多肽(PP)或其类似物；
-胰高血糖素受体激动剂或拮抗剂，葡萄糖依赖性促胰岛素多肽(GIP)受体激动剂或拮抗剂，胃饥饿素拮抗剂或反向激动剂，Xenin及其类似物；
-二肽基肽酶-IV(DPP-4)抑制剂，例如：阿格列汀(例如 )，利格列汀(例如)，沙格列汀(例如Komboglyze )，西格列汀(例如 )，阿格列汀(anagliptin)，替格利汀(teneligin)(例如)，曲格列汀(trelagliptin)，维格列汀(例如)，吉格列汀(gemigliptin)，奥格列汀(omarigliptin)，依格列汀(evogliptin)，度格列汀(dutogliptin)，DA-1229，MK-3102，KM-223，KRP-104，PBL-1427，盐酸Pinoxacin，和Ari-2243；
-钠依赖性葡萄糖转运蛋白2(SGLT-2)抑制剂，例如：坎格列净(canagliflozin)，达格列净(dapagliflozin)，remogliflozin，remogliflozin etabonate，sergliflozin，empagliflozin，ipragliflozin，托格列净(tofogliflozin)，鲁格列净(luseogliflozin)，埃格列净(ertugliflozin)，EGT-0001442，LIK-066，SBM-TFC-039，和KGA-3235(DSP-3235)；
-SGLT-2和SGLT-1的双重抑制剂(例如LX-4211，LIK066)。
-SGLT-1抑制剂(例如LX-2761，KGA-3235)或SGLT-1抑制剂与抗肥胖症药物例如回肠胆汁酸转运(IBAT)抑制剂(例如GSK-1614235+GSK-2330672)组合：
-双胍类(例如二甲双胍，丁双胍，苯乙双胍)；
-噻唑烷二酮类(例如吡格列酮，罗格列酮)，格列酮类似物(例如洛格列酮(lobeglitazone))；
-过氧化物酶体增殖物激活受体(PPAR-)(α，γ或α/γ)激动剂或调节剂(例如索格列扎(saroglitazar)(例如)，GFT-505)或PPARγ部分激动剂(例如Int-131)；
-磺酰脲类(例如甲苯磺丁脲，格列本脲，格列美脲，格列吡嗪)和氯茴苯酸类(例如那格列奈，瑞格列奈，米格列奈)；
-α-葡萄糖苷酶抑制剂(例如阿卡波糖，米格列醇，伏格列波糖)；
-胰淀素和胰淀素类似物(例如普兰林肽，)；
-G-蛋白偶联受体119(GPR119)激动剂(例如GSK-1292263，PSN-821，MBX-2982，APD-597，ARRY-981，ZYG-19，DS-8500，HM-47000，YH-Cheml)；
-GPR40激动剂(例如TUG-424，P-1736，P-11187，JTT-851，GW9508，CNX-011-67，AM-1638，AM-5262)；
-GPR120激动剂和GPR142激动剂；
-全身或低吸收性TGR5(GPBAR1＝G-蛋白偶联胆汁酸受体1)激动剂(例如INT-777，XL-475，SB756050)；
-糖尿病免疫治疗剂，例如：口服C-C趋化因子受体2型(CCR-2)拮抗剂(例如CCX-140，JNJ-41443532)，白介素1β(IL-1β)拮抗剂(例如AC-201)，或口服单纯系抗体(MoA)(例如methalozamide，VVP808，PAZ-320，P-1736，PF-05175157，PF-04937319)；
-用于治疗代谢综合征和糖尿病的抗炎剂，例如：核因子κB抑制剂(例如)；
-腺苷单磷酸激活的蛋白激酶(AMPK)刺激剂，例如：Imeglimin(PXL-008)，Debio-0930(MT-63-78)，R-118；
-11-β-羟基类固醇脱氢酶1(11-β-HSD-1)抑制剂(例如LY2523199，BMS770767，RG-4929，BMS816336，AZD-8329，HSD-016，BI-135585)；
-葡萄糖激酶活化剂(例如PF-04991532，TTP-399(GK1-399)，GKM-001(ADV-1002401)，ARRY-403(AMG-151)，TAK-329，TMG-123，ZYGK1)；
-甘油二酯O-酰基转移酶(DGAT)抑制剂(例如pradigastat(LCQ-908))，蛋白质酪氨酸磷酸酶1抑制剂(例如trodusquemine)，葡萄糖-6-磷酸酶抑制剂，果糖-1，6-二磷酸酶抑制剂，糖原磷酸化酶抑制剂，磷酸烯醇丙酮酸羧激酶抑制剂，糖原合酶激酶抑制剂，丙酮酸脱氢酶激酶抑制剂；
-葡萄糖转运蛋白-4调节剂，生长抑素受体3激动剂(例如MK-4256)；
-一种或多种降脂剂也适于作为组合搭档，例如：3-羟基-3-甲基戊二酰基辅酶A还原酶(HMG-CoA-还原酶)抑制剂，如辛伐他汀(例如 )，阿托伐他汀(例如)，瑞舒伐他汀(例如)，普伐他汀(例如)，氟伐他汀(例如)，匹伐他汀(例如)，洛伐他汀(例如)，美伐他汀(例如)，利伐他汀(rivastatin)，西立伐他汀贝特类，如苯扎贝特(例如延缓(retard))，环丙贝特(例如)，非诺贝特(例如)，吉非贝齐(gemfibrozil)(例如)，依托贝特(etofibrate)，双贝特(simfibrate)，罗尼贝特(ronifibrate)，克利贝特(clinofibrate)，氯贝胺(clofibride)，烟酸及其衍生物(例如烟酸，包括烟酸的缓释制剂)，烟酸受体1激动剂(例如GSK-256073)，PPAR-δ激动剂，乙酰-CoA-乙酰转移酶(ACAT)抑制剂(例如阿伐麦布(avasimibe))，胆固醇吸收抑制剂(例如依折麦布(ezetimibe)，S-556971)，胆汁酸结合物质(例如考来烯胺，考来维仑)，回肠胆汁酸转运(IBAT)抑制剂(例如GSK-2330672，LUM-002)，微粒体甘油三酯转移蛋白(MTP)抑制剂(例如lomitapide(AEGR-733)，SLx-4090，granotapide)，前体蛋白转化酶枯草杆菌蛋白酶/kexin 9型(PCSK9)调节剂(例如alirocumab(REGN727/SAR236553)，AMG-145，LGT-209，PF-04950615，MPSK3169A，LY3015014，ALD-306，ALN～PCS，BMS-962476，SPC5001，ISIS-394814，1B20，LGT-210，1D05，BMS-PCSK9Rx-2，SX-PCK9，RG7652)，LDL受体上调调节剂，例如肝选择性甲状腺激素受体β激动剂(例如伊罗替罗(eprotirome)(KB-2115)，MB07811，sobetirome(QRX-431)，VIA-3196，ZYT1)，升HDL化合物如：胆固醇酯转移蛋白(CETP)抑制剂(例如anacetrapib(MK0859)，达塞曲匹(dalcetrapib)，evacetrapib，JTT-302，DRL-17822，TA-8995，R-1658，LY-2484595，DS-1442)，或双重CETP/PCSK9抑制剂(例如K-312)，ATP结合盒(ABC1)调节剂，脂质代谢调节剂(例如BMS-823778，TAP-301，DRL-21994，DRL-21995)，磷脂酶A2(PLA2)抑制剂(例如达拉帕地(darapladib)，varespladib，瑞拉帕地(rilapladib))，ApoA-I增强剂(例如RVX-208，CER-001，MDCO-216，CSL-112)，胆固醇合成抑制剂(例如ETC-1002)，脂质代谢调节剂(例如BMS-823778，TAP-301，DRL-21994，DRL-21995)和ω-3脂肪酸及其衍生物(例如二十碳五烯酸乙基(icosapent ethyl)(AMR101)，AKR-063，NKPL-66，PRC-4016，CAT-2003)；
-溴隐亭(-bromocriptine)(例如)，苯丁胺和苯丁胺制剂或组合(例如Adipex-P，Ionamin，)，苄非他明(benzphetamine)(例如)，二乙胺苯丙酮(diethylpropion)(例如)，苯二甲吗啉(phendimetrazin)(例如)，安非他酮及组合(例如Wellbutrin)，西布曲明(例如)，托吡酯(topiramat)(例如)，唑尼沙胺(zonisamid)(例如)，特索芬辛，阿片样物质拮抗剂如纳曲酮(例如纳曲酮+安非他酮)，大麻素受体1(CB1)拮抗剂(例如TM-38837)，黑色素聚集激素(MCH-1)拮抗剂(例如BMS-830216，ALB-127158(a))，MC4受体激动剂和部分激动剂，RM-493)，神经肽Y5(NPY5)或NPY2拮抗剂(例如velneperit，S-234462)，NPY4激动剂(例如PP-1420)，β-3-肾上腺素能受体激动剂，瘦蛋白或瘦蛋白模拟物，5-羟基色胺2c(5HT2c)受体激动剂(例如氯卡色林(lorcaserin)，)，普兰林肽/美曲普汀，脂肪酶抑制剂如西替利司他(cetilistat)(例如)，奥利司他(例如)，血管生成抑制剂(例如ALS-L1023)，β组胺H3拮抗剂(例如HPP-404)，AgRP(agouti相关蛋白)抑制剂(例如TTP-435)，血清素再摄取抑制剂如氟西汀(例如)，度洛西汀(例如)，双重或三重单胺摄取抑制剂(多巴胺，去甲肾上腺素和血清素再摄取)如舍曲林(例如)，特索芬辛，甲硫氨酸氨肽酶2(MetAP2)抑制剂(例如beloranib)，和针对成纤维细胞生长因子受体4(FGFR4)(例如ISIS-FGFR4Rx)或阻抑素(prohibitin)靶向肽-1(例如)的产生的反义寡核苷酸；
-一氧化氮供体，AT1拮抗剂或血管紧张素II(AT2)受体拮抗剂如替米沙坦(例如)，坎地沙坦(例如)，缬沙坦(例如)，氯沙坦(losartan)(例如)，依普沙坦(例如)，厄贝沙坦(irbesartan)(例如)，奥美沙坦(例如)，他索沙坦，阿齐沙坦(例如)，双重血管紧张素受体阻断剂(双重ARB)，血管紧张素转化酶(ACE)抑制剂，ACE-2激活剂，肾素抑制剂，肾素原抑制剂，内皮素转化酶(ECE)抑制剂，内皮素受体(ET1/ETA)阻断剂，内皮素拮抗剂，利尿剂，醛固酮拮抗剂，醛固酮合酶抑制剂，α-阻断剂，α-2肾上腺素能受体拮抗剂，β-阻断剂，α-/β-混合阻断剂，钙拮抗剂，钙通道阻断剂(CCB)，钙通道阻断剂地尔硫的鼻制剂(例如CP-404)，双盐皮质激素/CCB，中枢作用的抗高血压药，中性肽链内切酶抑制剂，氨基肽酶A抑制剂，血管肽(vasopeptide)抑制剂，双重血管肽抑制剂如脑啡肽酶-ACE抑制剂或脑啡肽酶-ECE抑制剂，双重作用AT受体-脑啡肽酶抑制剂，双重AT1/ETA拮抗剂，晚期糖基化终产物(AGE)破坏剂，重组肾酶，血压疫苗如抗RAAS(肾素-血管紧张素-醛固酮系统)疫苗，AT1-或AT2-疫苗，基于高血压药物基因组学的，如具有抗高血压响应的遗传多态性调节剂，血小板聚集抑制剂，以及其他或其组合是合适的。
“核酸分子”是脱氧核糖核酸(DNA)或核糖核酸(RNA)。核酸分子可以是单链或双链及线性或共价闭合以形成环的分子形式。
术语“DNA”涉及包含脱氧核糖核苷酸残基且通常完全或基本上由脱氧核糖核苷酸残基组成的分子。“脱氧核糖核苷酸”涉及在β-D-呋喃核糖基的2′-位缺少羟基的核苷酸。术语“DNA”包括分离的DNA，例如部分或完全纯化的DNA，基本上纯的DNA，合成的DNA和重组产生的DNA，并且包括通过添加，缺失，取代和/或改变一个或多个核苷酸而与天然存在的DNA不同的修饰的DNA。这种改变可以包括添加非核苷酸材料，例如添加至DNA的末端或内部，例如添加于DNA的一个或多个核苷酸处。DNA分子中的核苷酸还可以包含非标准核苷酸，例如非天然存在的核苷酸或化学合成的核苷酸。这些改变的DNA可以成为类似物或天然存在的DNA的类似物。当与核苷酸结合使用时，术语“天然存在的”是指碱基腺嘌呤(A)，胞嘧啶(C)，鸟嘌呤(G)，胸腺嘧啶(T)和尿嘧啶(U)。
术语“RNA”涉及包含核糖核苷酸残基并且通常完全或基本上由核糖核苷酸残基组成的分子。“核糖核苷酸”涉及在β-D-呋喃核糖基的2′-位具有羟基的核苷酸。术语“RNA”包括分离的RNA，例如部分或完全纯化的RNA，基本上纯的RNA，合成的RNA和重组产生的RNA，并且包括通过添加，缺失，取代和/或改变一个或多个核苷酸而与天然存在的RNA不同的修饰的RNA。这种改变可以包括添加非核苷酸材料，例如添加至RNA的末端或内部，例如添加于RNA的一个或多个核苷酸处。RNA分子中的核苷酸还可以包含非标准核苷酸，例如非天然存在的核苷酸或化学合成的核苷酸或脱氧核苷酸。这些改变的RNA可以称为类似物或天然存在的RNA的类似物。根据本发明，“RNA”是指单链RNA或双链RNA。在一个实施方案中，RNA是mRNA，例如体外转录的RNA(IVT RNA)或合成的RNA。RNA也可以是修饰的，例如具有一个或多个增加RNA的稳定性(例如半衰期)的修饰。此类修饰是本领域技术人员已知的，并且包括例如5’-帽或5’帽类似物。
根据本发明的核酸分子可以包括/包含在载体中。本文所用的术语“载体”包括本领域技术人员已知的所有载体，包括质粒载体，粘粒载体，噬菌体载体(例如λ噬菌体)，病毒载体(如腺病毒或杆状病毒载体)或人工染色体载体，如细菌人工染色体(BAC)，酵母人工染色体(YAC)或P1人工染色体(PAC)。所述载体包括表达以及克隆载体。表达载体包括质粒以及病毒载体，并且通常含有期望的编码序列和在特定宿主生物体(例如细菌，酵母，植物，昆虫或哺乳动物)中或在体外表达系统中表达可操作连接的编码序列所必需的适当DNA序列。克隆载体通常用于工程化和扩增某些期望的DNA片段，并且可以缺少表达期望的DNA片段所需的功能性序列。
或者，根据本发明的核酸分子可以整合到基因组中，例如宿主细胞的基因组中。将特定核酸分子整合到基因组中的方法和方法是本领域技术人员已知的。
术语“细胞”或“宿主细胞”通常涉及完整细胞，即具有完整膜且未释放其正常细胞内组分如酶、细胞器或遗传物质的细胞。完整细胞通常是活细胞，即能够发挥其正常代谢功能的活细胞。该术语通常涉及可以用外源核酸转染或转化的任何细胞。当用外源核酸转染或转化并转移至受体时，细胞通常可以在受体中表达所述核酸。术语“细胞”包括原核细胞，如细菌细胞和真核细胞，例如酵母细胞，真菌细胞或哺乳动物细胞。合适的细菌细胞包括来自如下的细胞：革兰氏阴性细菌菌株，如大肠杆菌(Escherichiacoli)、变形菌属(Proteus)和假单胞菌属(Pseudomonas)的菌株，和革兰氏阳性菌株，例如茅孢杆菌属(Bacillus)、链霉菌属(Streptomyces)，葡萄球菌属(Staphylococcus)和乳球菌属(Lactococcus)的菌株。合适的真菌细胞包括来自木霉属(Trichoderma)，脉孢菌属(Neurospora)和曲霉属(Aspergillus)的细胞。合适的酵母细胞包括来自酵母属(Saccharomyces)(例如酿酒酵母(Saccharomyces cerevisiae))，裂殖酵母属(Schizosaccharomyces)(例如粟酒裂殖酵母(Schizosaccharomyces pombe))，毕赤酵母(Pichia)(例如巴斯德毕赤酵母(Pichiapastoris)和甲醇毕赤酵母(Pichia methanolica))和汉逊酵母属(Hansenula)的细胞。合适的哺乳动物细胞包括例如CHO细胞，BHK细胞，HeLa细胞，COS细胞，HEK293等。在一个实施方案中，使用HEK293细胞。然而，也可以使用本领域中用于表达异源蛋白的两栖动物细胞、昆虫细胞、植物细胞和任何其它细胞。哺乳动物细胞，如哺乳动物脂肪细胞，尤其用于接受性转移(adoptive transfer)，包括来自人，小鼠，仓鼠，猪，山羊和灵长类动物的细胞。细胞可以衍生自许多种组织类型并且包括原代细胞和细胞系，如免疫系统的细胞，特别是抗原呈递细胞例如树突状细胞和T细胞，干细胞如造血干细胞和间充质干细胞及其他细胞类型。抗原呈递细胞是在其表面上主要组织相容性复合体的背景下显示抗原的细胞。T细胞可以使用其T细胞受体(TCR)识别该复合物。“细胞”或“宿主细胞”可以是分离的或组织或生物体(特别是“非人生物体”)的一部分。
本文所用的术语“非人生物体”意在包括非人灵长类动物或其他动物，特别是哺乳动物，例如牛，马，猪，绵羊，山羊，狗，猫，兔或啮齿动物，如小鼠，大鼠，豚鼠和仓鼠。
本发明中表征的药物组合物通常含有治疗有效量的人FGF21的变体，任选地作为本文所述的融合分子、核酸分子或宿主细胞(本文也称为“剂(agents)”)的一部分，以产生期望的反应或期望的效果。
根据本发明的药物组合物还可以包含至少一种其它活性药物成分，例如至少一种如上文定义的其它活性药物成分。
根据本发明的药物组合物通常是无菌组合物。药物组合物通常以均匀剂型提供，并且可以以本身已知的方式制备。药物组合物可以例如为溶液或悬浮液的形式。
药物组合物可以进一步包含一种或多种载剂和/或赋形剂，所有这些都是医药上可接受之。本文所用的术语“医药上可接受之”是指不与药物组合物的活性成分的作用相互作用的材料的无毒性。
术语“载剂”是指天然或合成性质的有机或无机组分，其中活性组分组合以便于促进，增强应用或使得能够应用。根据本发明，术语“载体”还包括适于施用于受试者的一种或多种相容的固体或液体填充剂、稀释剂或包封物质。
用于肠胃外给药的可能的载体物质是例如无菌水，林格氏溶液(Ringer’ssolution)，乳酸化林格氏溶液，生理盐水，抑菌盐水(例如含有0.9％苄醇的盐水)，磷酸盐缓冲盐水(PBS)，Hank’s溶液，聚烷撑乙二醇类(polyalkylene glycols)，氢化萘类，特别是生物相容的丙交酯聚合物，丙交酯/乙交酯共聚物或聚氧乙烯/聚氧丙烯共聚物。
本文所用的术语“赋形剂”意在包括可以存在于药物组合物中且不是活性成分的所有物质，例如盐，粘合剂(例如乳糖，右旋糖，蔗糖，海藻糖，山梨醇，甘露醇)，润滑剂，增稠剂，表面活性剂，防腐剂，乳化剂，缓冲物质，调味剂或着色剂。
不是医药上可接受之盐可以用于制备医药上可接受之盐，并且包括在本发明中。这种类型的医药上可接受之盐以非限制性方式包括由如下的酸制备的那些：盐酸，氢溴酸，硫酸，硝酸，磷酸，马来酸，乙酸，水杨酸，柠檬酸，甲酸，丙二酸，琥珀酸等。医药上可接受之盐也可以制备为碱金属盐或碱土金属盐，例如钠盐，钾盐或钙盐。可以加入盐以调节离子强度或张力。
用于药物组合物的合适的防腐剂包括抗氧化剂，柠檬酸，柠檬酸钠，苯扎氯铵，氯丁醇，半胱氨酸，甲硫氨酸，对羟基苯甲酸酯类和硫柳汞。
用于药物组合物的合适的缓冲物质包括盐中的乙酸，盐中的柠檬酸，盐中的硼酸和盐中的磷酸。
药物组合物还可以配制为稳定的冻干产品，其用合适的稀释剂重建，所述稀释剂任选地包含一种或多种如上所定义的赋形剂。
如本文所使用的，术语“组件试剂盒(简称：试剂盒)”是指包括一个或多个容器和任选的数据载体的制品。所述一个或多个容器可以填充有一种或多种上述(试)剂。试剂盒中可包括另外的容器，其含有例如稀释剂，缓冲液和其它试剂。所述数据载体可以是非电子数据载体，例如图形数据载体，例如信息页，信息表单，条形码或访问码，或电子数据载体，例如光盘(CD)数字通用光盘(DVD)，微芯片或另一基于半导体的电子数据载体。访问代码可以允许访问数据库，例如互联网数据库，集中式或分散式数据库。所述数据载体可包括用于使用本发明表征的FGF21变体的说明书。
本文所述的药剂和组合物可以通过任何常规途径施用，例如口服，肺，吸入或胃肠外途径，包括通过注射或输注。在一个实施方案中，使用肠胃外施用，例如静脉内，动脉内，皮下，皮内或肌内。本文所述的药剂和组合物也可以通过缓释施用来施用。
适于肠胃外给药的药物组合物通常包含活性化合物的无菌水性或非水性制备物，其通常与接受者的血液等渗。相容的载剂/溶剂/稀释剂的实例是无菌水，林格氏溶液，乳酸林格氏溶液，生理盐水，抑菌盐水(例如含有0.9％苄醇的盐水)，磷酸盐缓冲盐水(PBS)和Hank′s溶液。此外，通常无菌的固定油可以用作溶液或悬浮介质。
本文所述的药剂和组合物通常以治疗有效量施用。“治疗有效量”是指单独或与其它剂一起达到期望的治疗反应或望的治疗效果的量，通常不会引起不可接受的副作用。在治疗具体的疾病或具体病症的情况下，期望的反应通常涉及抑制疾病的进程。这包括减缓疾病的进展，特别是中断或逆转疾病的进展。在疾病或病症的治疗中期望的反应还可以是延迟所述疾病或所述病症的发作或预防所述疾病或所述病症的发作。本文所述的药剂或组合物的有效量将取决于要治疗的病况，疾病的严重程度，受试者的个人参数，包括年龄，生理状况，体格和体重，治疗持续时间，伴随治疗的类型(如果有的话)，具体施用途径和类似因素。因此，本文所述的药剂的施用剂量可取决于各种此类参数。在受试者中采用初始剂量的反应不足的情况下，可以使用更高的剂量(或通过不同的，更局部的施用途径实现的有效更高剂量)。
本发明还提供了本发明的人FGF21的变体与至少一种其它活性药物成分的组合。
在一个实施方案中，本发明的人FGF21的变体与至少一种其它活性药物成分的组合可以通过将活性药物成分分别施用于患者或以组合产品的形式来应用，所述组合产品中多种活性药物成分存在于一种药物组合物中。当分别施用时，施用可以同时进行或以任何顺序来序贯进行。对FGF21变体和其它活性药物成分的量及相对给药时间进行选择以实现期望的组合治疗效果。组合的施用可以同时(concomitantly)呈：(1)包括所有活性药物成分的单一药物组合物；或(2)分别包含至少一种活性药物成分的多种药物组合物。或者，所述组合可以以序贯方式分开施用，其中首先施用一种治疗剂，然后施用另一第二治疗剂，或反之。这种序贯施用可以在时间上间隔近或远。
根据本发明，术语“疾病或病症”意指任何病理学或非健康状态，特别是肥胖症，超重，代谢综合征，糖尿病，高血糖症，血脂异常，非酒精性脂肪性肝炎(NASH)和/或动脉粥样硬化。
术语“肥胖症”是指其中过量体脂肪积累到可能对健康具有负面影响的程度的医学状况。就人(成人)受试者而言，肥胖症可定义为大于或等于30kg/m2(BMI≥30kg/m2)的体重指数(BMI)。
术语“超重”是指其中身体脂肪的量高于最佳健康的医学状况。就人(成人)受试者而言，肥胖症可定义为大于或等于25kg/m2(例如25kg/m2≤BMI＜30kg/m2)的体重指数(BMI)。
BMI是高度重量的简单指数，其通常用于对成人中的超重和肥胖症进行分类。其定义为以千克计的人的体重除以以米计的身高的平方(kg/m2)。
“代谢综合征”可以定义为以下医学状况中的至少三种的聚群：腹部(中心)肥胖症(例如，对于白人男性定义为腰围≥94cm，对于白人女性定义为≥80cm，其它组具有种族特异性值)，升高的血压(例如130/85mmHg或更高)，升高的空腹血浆葡萄糖(例如至少100mg/dL)，血清甘油三酯高(例如至少150mg/dL)和高密度脂蛋白(HDL)水平低(例如对于雄性小于40mg/dL和对于雌性小于50mg/dL)。
“糖尿病”(也简称为“糖尿病”)是指一类代谢疾病，其特征在于高水平的血糖，其由胰岛素生产缺陷，胰岛素作用缺陷或二者导致。在一个实施方案中，糖尿病选自1型糖尿病，2型糖尿病，妊娠糖尿病，成人晚发性自身免疫性糖尿病(LADA)，年轻人成熟发作型糖尿病(maturity onset diabetes ofthe young，MODY)和由于特定的遗传状况，药物，营养不良，感染和其他疾病导致的其他类型的糖尿病。
目前世界卫生组织糖尿病诊断标准如下：空腹血浆葡萄糖≥7.0mmol/1(126mg/dL)或2-h血浆葡萄糖≥11.1mmol/l(200mg/dL)。
“1型糖尿病”(也称为“胰岛素依赖型糖尿病(IDDM)”或“青少年糖尿病”)是这样的病症，其特征为由完全缺乏胰岛素引起的高血糖水平。这发生在身体的免疫系统攻击胰腺中产生胰岛素的β细胞并破坏它们时。然后，胰腺产生很少的胰岛素或不产生胰岛素。胰腺移除或疾病也可导致胰岛素产生的β细胞的丢失。1型糖尿病占糖尿病病例的5％至10％。
“2型糖尿病”(也称为“非胰岛素依赖性糖尿病(NIDDM)”或“成人型糖尿病”)是这样的病症，其特征为过量葡萄糖产生而不论胰岛素可用性，并且循环葡萄糖水平保持过高，其原因是葡萄糖清除不足(胰岛素作用)。2型糖尿病可占所有诊断的糖尿病病例的约90％至95％。
“妊娠糖尿病”是没有既往诊断出的糖尿病的妇女在怀孕期间(特别是在妊娠末期)显示出高血糖水平的状况。妊娠糖尿病影响3-10％的妊娠，取决于研究的人群。
“成人迟发性自身免疫性糖尿病(LADA)”(也称为“慢性1型糖尿病”)是在成年人中发生的一种形式的1型糖尿病，通常具有较慢的发作过程。
“年轻人成熟型糖尿病(MODY)”是指由常染色体显性基因突变(其破坏胰岛素生成)导致的遗传形式的糖尿病。
术语“高血糖症”是指血液中糖(葡萄糖)过量。
术语“血脂异常”是指脂蛋白代谢的病症，包括脂蛋白过量生成(“高脂血症”)或缺乏(“低脂血症”)。血脂异常可通过血液中总胆固醇，低密度脂蛋白(LDL)胆固醇和/或甘油三酯浓度升高和/或高密度脂蛋白(HDL)胆固醇浓度降低来显示。
非酒精性脂肪性肝炎(NASH)是一种肝脏疾病，其特征在于脂肪(脂滴)的累积，以及肝细胞的炎症和变性。一旦患上，该疾病伴有肝硬化的高风险，其是一种肝功能改变并可进展为肝功能不全的状态。此后，NASH常常进展为肝癌。
“动脉粥样硬化”是一种血管疾病，其特征为大型和中型动脉内膜中不规则分布的脂质沉积，称为斑块，其可导致动脉管腔变窄并发展成纤维化和钙化。病变通常是局灶性的，进展缓慢的和间歇性的。偶尔发生斑块破裂导致血流阻塞，引起阻塞远端的组织死亡。血流的受限占了临床表现的大多数，其随阻塞的分布和严重程度而变化。
本文所用的术语“药物”是指用于治疗，即用于治疗疾病或病症的物质/组合物。
“治疗”意指对受试者施用化合物或组合物或化合物或组合物的组合，以预防或消除疾病或病症；阻滞或减缓受试者的疾病或病症；抑制或减缓受试者中新疾病或病症的发展；降低目前患有或先前患有疾病或病症的受试者中的症状和/或复发的频率或严重程度；和/或延长，即增加受试者的寿命。
具体地，术语“治疗/治疗疾病或病症”包括治愈疾病或病症或其症状，缩短疾病或病症或其症状的持续时间，改善、预防疾病或病症或其症状、减缓或抑制疾病或病症或其症状的进展或恶化，或预防或延迟疾病或病症或其症状的发作。
根据本发明，术语“受试者”是指用于治疗的受试者，具体而言是患病受试者(也称为“患者”)，包括人，非人灵长类动物或其他动物，特别是哺乳动物，例如牛，马，猪，绵羊，山羊，狗，猫，兔或啮齿动物，如小鼠，大鼠，豚鼠和仓鼠。在一个实施方案中，受试者/患者是人。
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实施例
实施例1：HEK293细胞或大肠杆菌(E.coli)中FGF21变体的表达
通过瞬时转染HEK293细胞或在大肠杆菌中产生FGF21变体。
(a)HEK293细胞中FGF21变体的表达
将FGF21变体的DNA序列以N端融合至IL2信号序列(SEQ ID NO：4)，随后是富含组氨酸的序列(His标签)和TEV蛋白酶切割位点(SEQ ID NO：5或6)。信号序列是目的蛋白分泌到培养基中所需的。使用固定化金属离子亲和色谱(IMAC)(完全His-标签纯化柱，Roche)从培养物上清液中纯化目的蛋白。从IMAC柱洗脱后，任选地通过添加TEV蛋白酶来切割N-末端His-标签。在His-标签切割之后，将切割反应溶液第二次通过固定化金属亲和柱(IMAC-柱)(完全His-标签纯化柱，Roche)，收集(无His标签地)流出物级分。使用采用磷酸盐缓冲盐水(PBS，Gibco)作为运行缓冲液的凝胶过滤柱进一步纯化蛋白质。收集含有目的蛋白的级分，合并，浓缩并储存在-80℃直至进一步使用。
(b)大肠杆菌中FGF21变体的表达
SEQ ID NO：2(成熟人野生型FGF21，即没有信号序列；也称为FGF21H29-S209)和SEQ ID NO：3(成熟人野生型FGF21，具有额外的N-末端G；也称为G+FGF21H29-S209或G-FGF21H29-S209或简称为G-FGF21)的FGF21蛋白以及SEQ ID NO：124至SEQ ID NO：157和SEQID NO：174的蛋白在大肠杆菌中表达。FGF21蛋白的DNA序列以N-末端融合至富含组氨酸的序列(His-标签)和TEV或SUMO蛋白酶切割位点(SEQ ID NO：6或7)。使用固定化的金属离子亲和色谱(IMAC)(His Trap HP，GE Healthcare)纯化目的蛋白，然后通过加入TEV或SUMO蛋白酶裂解N-末端His-标签。在His-标签裂解后，使用离子交换柱纯化裂解反应溶液，然后使用磷酸盐缓冲盐水(PBS，Gibco)作为运行缓冲液进行凝胶过滤步骤。收集含有目的蛋白的级分，合并，浓缩并储存在-80℃直至进一步使用。
实施例2：人和鼠血浆中FGF21变体的稳定性测量
小鼠和大鼠中成熟人野生型FGF21的体内药代动力学(PK)分析揭示了小于1小时的半衰期，这是由于通过肾的快速清除(Kharitonenkov A.等(2007)Endocrinology 148(2)：774-781；Stein S.等(2009)Diabetes Care 32(1)：126-8；Lin Z.等(2011)PLoS ONE6(4)：e18398)。另外，还发现FGF21对体内蛋白水解降解敏感。这种降解显著地负面影响FGF21的治疗潜力(表1)(Hecht R.等(2012)PLoS ONE 7(11)：e49345；Micanovic R.等(2009)J Cell Physiol.219(2)：227-34；Yie J.等(2012)Chem Biol Drug Des.79(4)：398-410)。
表1：FGF21的C末端氨基酸的丢失降低其体外活性
为了更多地了解人FGF21的代谢稳定性并鉴定由血液中蛋白水解降解形成的代谢物，如下所述分析蛋白质。具体而言，将15μmol/L的人野生型FGF21(SEQ ID NO：3)在人、小鼠和大鼠肝素锂血浆中于37℃体外温育60分钟。在温育结束时，根据制造商描述的程序用Norgen血清消耗试剂盒(Norgen Biotek，Thorold，ON，Canada，#17300)处理样品。从血浆样品中消耗掉主要血清蛋白，包括白蛋白，α-抗胰蛋白酶，转铁蛋白和触珠蛋白。在4℃，将50μL血浆样品施加于活化的离心(spin)柱上，并在台式微量离心机中以6,700x g离心1分钟，其中弃去流出液。通过加入500μL洗涤缓冲液将柱洗涤两次并离心1分钟。向柱中加入两次100μL洗脱缓冲液，离心1分钟以洗脱结合的蛋白。
通过在质谱范围为m/z 650至m/z 1700的离子阱质谱仪上，在正电离模式用精确质量测量(Thermo-Fisher LTQ-Orbitrap，NanoLC 1D Ultra，AS2自动进样器)通过液相色谱-质谱(LC-MS)实验来鉴定代谢物。用于代谢物鉴定的HPLC条件如下：柱Vydac MS C4300A，5μm，150mm x 75μm(Grace，Lokeren，Belgium)；流动相：洗脱液A 0.1％甲酸溶液伴随2％(v/v)的乙腈和洗脱液B乙腈+0.1％甲酸；运行时间75分钟；流速：280nL/min。通过Magtran软件(Zhongqi Zhang)将多电荷质量解卷积，并且通过GPMAW程序(LighthouseData，Odense，Denmark)显示形成的代谢物的蛋白序列。
如上所述SEQ ID NO：3的FGF21在血浆中温育1小时产生了快速切割，可检测到几种代谢物，其列于表2中。在所研究的所有物种的血浆中形成了对应于人野生型FGF21(SEQID NO：1)的位置29位至位置199的氨基酸序列的切割产物。该代谢物可以描述为具有18428.56Da的分子量的人-FGF21-des-十肽。SEQ ID NO：3的人FGF21及其切割产物的质谱和去卷积示于图1中。
表2：在人、大鼠和小鼠的血浆中于1小时温育后鉴定出的人FGF21的代谢物(氨基酸的编号根据SEQ ID NO：1的人野生型FGF21)
人
大鼠
小鼠
通过在人或小鼠血浆中于37℃温育蛋白多至24小时，以及用夹心免疫测定法分析剩余的完整全长FGF21蛋白，从而分析成熟人野生型FGF21(SEQ ID NO：2)和各种FGF21变体的体外稳定性。具体而言，将成熟人FGF21(SEQ ID NO：2)或变体以100ng/mL的浓度添加到血液样品(K2EDTA血浆，数个健康人供体的混合物)中，并于37℃温育多至24小时。在0，1，8和/或24小时后，取样，并通过加入2x浓缩的蛋白酶抑制剂(Protease Inhibitor CocktailTablets，Roche)终止温育。然后将样品转移至-80℃直至进一步处理。随后，使用人完整FGF21 ELISA(F2131-K01，Eagle Biosciences，Nashua，NH，USA)来评估剩余的完整全长FGF21蛋白的量。该测定法利用夹心技术，其中两种选择的抗体特异性结合至成熟人FGF21(aa1-7)的N-末端和C-末端(aa175-181)，如由供应商所描述的，其对应于全长人野生型FGF21(SEQ ID NO：1)的氨基酸aa29-35和aa203-209。每个单独样品在温育开始时的浓度用作内部参照且其与在稍后时间点处测得的浓度相关。
测试的重组FGF21变体在人和鼠血浆中的稳定性的结果总结在表3中，代表性的结果显示在图2中。
表3：FGF21突变对其针对蛋白水解的稳定性的影响(氨基酸的编号基于SEQ IDNO：1的人野生型FGF21)。
实施例3：在CHO细胞中针对人FGF21受体功效的体外细胞测定(In-Cell Western)
使用特异性和高度灵敏的In-Cell Western(ICW)测定法来测量成熟人FGF21(SEQID NO：2)或FGF21变体的细胞体外功效。ICW测定是通常以微孔板形式进行的免疫细胞化学测定。将稳定表达人FGFR1c以及人β-Klotho(KLB)的CHO Flp-In细胞(Invitrogen，Darmstadt，Germany)用于使用In-Cell Westem的FGF21受体自磷酸化测定(Aguilar H.N.等(2010)PLoS ONE 5(4)：e9965)。为了确定受体自磷酸化水平或MAP激酶ERK1/2的下游激活，将2×104个细胞/孔接种至96孔板中并生长48小时。细胞用无血清培养基Ham′s F-12营养混合物与GlutaMAX(Gibco，Darmstadt，Germany)血清饥饿3-4小时。随后用增加的浓度的成熟人FGF21(SEQ ID NO：2)、指定的FGF21变体或其他肽在37℃处理细胞5分钟。温育后，弃去培养基，将细胞在3.7％新鲜制备的多聚甲醛中固定20分钟。用0.1％Triton-X-100在PBS中的溶液将细胞透明化20分钟。用Odyssey封闭缓冲液(LICOR，Bad Homburg，Germany)在室温封闭2小时。加入抗pFGFR Tyr653/654(New England Biolabs，Frankfurt，Germany)或抗pERK磷酸-p44/42MAP激酶Thr202/Tyr204(Cell Signaling)作为一抗，并在4℃温育过夜。在温育一抗后，用加有0.1％Tween20的PBS洗涤细胞。将二抗-小鼠800CW抗体(LICOR，BadHomburg，Germany)在室温温育1小时。随后，用PBS加0.1％Tween20再次洗涤细胞，并用Odyssey成像仪(LICOR，Bad Homburg，Germany)对红外染料信号进行定量。通过用TO-PRO3染料(Invitrogen，Karlsruhe，Germany)对DNA进行定量来将结果标准化。以任意单位(AU)获得数据，并且从剂量-响应曲线获得EC50值，并且总结在表4中。图3显示来自过表达人FGFR1c加KLB的CHO细胞的ICW的结果。
表4：在CHO细胞中通过ICW pERK测得的成熟人FGF21(SEQ ID NO：2)和FGF21变体的EC50值。除另有说明外，通过使用N-末端标签(SEQ ID NO：5或6)纯化测试的FGF21蛋白，即除另有说明外，在确定EC 50值时，FGF21蛋白携带N-末端标签。氨基酸的编号是指SEQ IDNO：1的人野生型FGF21。
实施例4：用原代人脂肪细胞对FGF21变体的体外活性测量
还使用In-Cell Western(ICW)测定以人原代皮下脂肪细胞测量了人FGF21(SEQID NO：2)或FGF21变体的细胞体外功效(Aguilar H.N.等(2010)PLoS ONE 5(4)：e9965)。简言之，将2.8×104个人前脂肪细胞(PromoCell，Heidelberg，Germany)接种到96孔板的每个孔中并分化成成熟脂肪细胞(Hemmrich K.等(2005)Differentiation 73(1)：28-35；LeeM.J.(2014)Methods Enzymol.538：49-65)。在MAP激酶ERK1/2激活的刺激之前，将细胞用无血清培养基(DMEM1g/l/Ham′s F-10培养基(1∶1，v/v)(PAN-Biotech，Aidenbach，Germany)，15mmol/L Hepes，pH7.4，33μmol/L生物素，17μmol/L泛酸盐)血清饥饿3-4小时。随后用渐增浓度的成熟人FGF21(SEQ ID NO：2)、指定的FGF21变体、或其他肽在37℃处理细胞5分钟。温育后，弃去培养基，将细胞在3.7％新鲜制备的多聚甲醛中固定20分钟。用0.1％Triton-X-100在PBS中的溶液将细胞透明化20分钟。用Odyssey封闭缓冲液(LICOR，Bad Homburg，Germany)在室温封闭2小时。加入抗pERK磷酸-p44/42MAP激酶Thr202/Tyr204(CellSignaling)作为一抗，并在4℃温育过夜。在温育一抗后，用加有0.1％Tween20的PBS洗涤细胞。将二抗-小鼠800CW抗体(LICOR，Bad Homburg，Germany)在室温温育1小时。随后，用PBS加0.1％Tween20再次洗涤细胞，并用Odyssey成像仪(LICOR，Bad Homburg，Germany)对红外染料信号进行定量。通过用TO-PRO3染料(Invitrogen，Karlsruhe，Germany)对DNA进行定量来将结果标准化。以任意单位(AU)获得数据，并且从剂量-响应曲线获得EC50值，如图4A和B所示。EC50值总结于表5中。
表5：在原代人脂肪细胞中通过ICW pERK测得的成熟人FGF21(SEQ ID NO：2)和FGF21变体的EC50值。氨基酸的编号依据SEQ ID NO：1的人野生型FGF21。
实施例5：人FGF21和FGF21变体的热稳定性分析
为了分析FGF21中的突变对热稳定性的影响，在用于搜索稳定化配体的差示扫描荧光测定(DSF或ThermoFluorTM)测定的模拟中应用了热迁移测定(Ahmad S.等(2012)Protein Science 21：433-446；Pantoliano等(2001)J.Biomol.Screen 6：429-440；Niesen等(2007)Nat.Protoc.2：2212-21)。该测定基于如下观察：疏水性荧光染料如SyproTMOrange(Life Technologies，目录号S6651)在它们结合蛋白上的疏水片层(patches)时增加它们的荧光。当这样的疏水片层在加热展开时暴露在蛋白质中，从而使荧光的增加可以用作展开程度的量度，并因此用于蛋白质的热稳定性。
通过将蛋白质在PBS(Gibco)中的溶液与SyproTM Orange(稀释于来自供应商提供的5000X DMSO储液的水中)的160X溶液混合来测试FGF21变体。用PBS将样品体积调节至20μl。通常的条件在最终混合物中含有0.8mg/ml FGF21变体蛋白和8X SyproTM Orange，但是蛋白质浓度可以在0.4mg/ml和1.2mg/ml之间变化。将样品分配于96孔PCR板(BioRad Semi-Skirt 96白色)中，并短暂离心以除去气泡。将平板插入BioRad iQ5实时PCR仪中，并以1℃/分钟的升温速度经历10至90℃的热梯度。对于荧光的激发和定量，选择波长485nm和575nm的滤光片。Biorad iQ5标准版软件(v.2.0.148.60623)用于数据处理。在荧光强度对温度的曲线中，选择拐点作为熔解温度(TM)的量度。在每个平板中包括人FGF21(SEQ ID NO：3)作为参照。图5显示选择的FGF21变体的代表性熔解曲线，表6列出相对于人FGF21(SEQ ID NO：3)的TM值差异。
表6：FGF21变体相对于SEQ ID NO：3的人FGF21的熔点差异(ΔTm)。括号中的值表示来自具有初始高荧光的曲线的数据，并且其较不可靠。“n.d.”：无法确定。测试的变体可以携带N末端His-标签(SEQ ID NO：5或6)。标签也可以通过蛋白酶切割去除。氨基酸的编号依据SEQ ID NO：1的人野生型FGF21。
实施例6：热稳定性变体设计及FGF21Q55C，N149C(SEQ ID NO：158)的X射线晶体结构
因为在像RCSB蛋白质数据库(PDB)这样的公共数据库中没有储存FGF21的结构，使用SWISS-MODEL同源性建模服务器(Kiefer等(2009)Nucleic Acids Res.37：D387-D392)或软件程序BioLuminate(BioLuminate，version 1.0，Schr6dinger，LLC，New York，NY，2012)建立了人FGF21(SEQ ID NO：2)的不同同源性模型。这些模型基于FGF19(Goetz等(2007)Mol.Cell.Biol.27：3417-3428；PDB码2P23；Harmer(2004)Biochemistry 43：629-640；PDB码1PWA)和FGF23(Goetz等(2007)Mol.Cell.Biol.27：3417-3428；PDB码2P39)的发表的晶体结构(图6)。使用BioLuminate对不同模型进行进一步分析以生成用于引入二硫键的潜在位点的列表。使用软件程序PYMOL(The PyMOL Molecular Graphics System，版本1.7LLC)在模型结构中目视检查潜在的位置，以最终选择用于表达和实验测试的候选物。
为了结晶人FGF21，如上所述(实施例1)表达和纯化经修饰和截短的变体(人FGF21S34-E176，Q55C，N149C；SEQ ID NO：158)。通过悬滴蒸汽扩散来生长晶体。将蛋白在50mmol/L Tris，pH 8.0，0.5mol/L NaCl中浓缩至12mg/mL。储存溶液含有2mol/L NH4SO4于100mmol/L Tris，pH 8.5中。将100nL蛋白溶液与100nL储存溶液混合并在20℃平衡。晶体在两周内出现。通过在补充有20至25％(体积/体积)乙二醇的储存溶液中浸泡晶体，然后在液氮中快速冷冻来实现冷冻保护。
在Swiss Light Source(SLS)in Viligen，Switzerland的光束线PX-III处收集X射线衍射数据，并用XDS(Kabsch W.(2010)Acta Crystallogr.D66：125-132)处理并用Aimless(Evans P.R.(2006)Acta Crystallogr.D62：72-82)进行衡量(scaled)，如autoProc(Vonrhein C.等(2011)Acta Crystallogr.D67：293-302)中实施的那样。晶体为空间群P4132，并且在不对称单元中含有两个FGF21Q55C，N149C分子。该晶体的晶胞尺寸如下：晶体衍射到分辨率，且Rmerge为10.5％。
使用程序Phaser(McCoy A.J.等(2007)J.Appl.Crystallogr.40：658-674)通过分子置换来确定结构。将FGF19(Protein Data Bank码1pwa)，FGF23(Protein Data Bank码2p39)和FGF9(Protein Data Bank码2P39)的晶体结构叠加，去除所有差异大于～的环。所得到的集合(ensemble)用作搜索模型，并产生明确的解。Phaser放置了三个FGF-集合。然而，第三集合的电子密度非常弱，且该集合被缺失。用Coot(Emsley P.等(2010)ActaCrystallogr.D66：486-501)进行建模，并用Buster(Bricogne G等(2011)Cambridge，United Kingdom：Global Phasing Ltd.)和Refmac5(Murshudov GN.等(2011)ActaCrystallogr.D67：355-367)进行细化。最终R因子为25.5％，游离R因子为28.6％。这些R因子相对较高，并且可以指示晶体中的一定量的无序。
在FGF21Q55C，N149C(SEQ ID NO：158)的最终模型中，分子A含有残基G42-K150和P158-G169；分子B含有残基P36-H154和P156-L170。在两个分子中，P150和R159之间的环是显著无序的，这指示了该区域的高度柔性(flexibility)。
可以假定这种高度柔性的环代表蛋白质的热展开的起点。因此，通过二硫键对该区域的稳定化可解释对于FGF21变体H29-S209 Q55C，N149C(SEQ ID NO：100)或相邻变体H29-S209 Q55C，P147C(SEQ ID NO：99)评估的热稳定性的约20℃的显著增加。
序列表
<110> 赛诺菲
<120> FGF21变体
<130> DE2015/152PCT
<150> EP 15 306 913.3
<151> 2015年12月2日
<160> 328
<170> PatentIn版本3.5
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<211> 209
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<213> 人(Homo sapiens)
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Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 3
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21
<400> 3
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 4
<211> 21
<212> PRT
<213> 人工序列
<220>
<223> IL2信号序列
<400> 4
Met Tyr Arg Met Gln Leu Leu Ser Cys Ile Ala Leu Leu Ser Leu Ala
1 5 10 15
Leu Val Thr Asn Ser
20
<210> 5
<211> 28
<212> PRT
<213> 人工序列
<220>
<223> His标签
<400> 5
Asp Ile Gln Thr Gly Gly Ser His Ala His Gly His Gly His Ala His
1 5 10 15
Gly His Gly Gly Ser Gly Glu Asn Leu Tyr Phe Gln
20 25
<210> 6
<211> 20
<212> PRT
<213> 人工序列
<220>
<223> His标签
<400> 6
Met Gly His His His His His His His His Gly Gly Gly Glu Asn Leu
1 5 10 15
Tyr Phe Gln Gly
20
<210> 7
<211> 108
<212> PRT
<213> 人工序列
<220>
<223> His标签 + SUMO标签
<400> 7
Met Gly His His His His His His Gly Ser Leu Gln Asp Ser Glu Val
1 5 10 15
Asn Gln Glu Ala Lys Pro Glu Val Lys Pro Glu Val Lys Pro Glu Thr
20 25 30
His Ile Asn Leu Lys Val Ser Asp Gly Ser Ser Glu Ile Phe Phe Lys
35 40 45
Ile Lys Lys Thr Thr Pro Leu Arg Arg Leu Met Glu Ala Phe Ala Lys
50 55 60
Arg Gln Gly Lys Glu Met Asp Ser Leu Arg Phe Leu Tyr Asp Gly Ile
65 70 75 80
Arg Ile Gln Ala Asp Gln Ala Pro Glu Asp Leu Asp Met Glu Asp Asn
85 90 95
Asp Ile Ile Glu Ala His Arg Glu Gln Ile Gly Gly
100 105
<210> 8
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S195C, S200C
<400> 8
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Cys Met Val Gly Pro Cys Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 9
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P199S, S200P
<400> 9
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Pro Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 10
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P199Q, Q201P
<400> 10
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Gln Ser Pro Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 11
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P199缺失的
<400> 11
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 12
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 M196P, V197G, G198V, P199M
<400> 12
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Pro Gly Val Met Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 13
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 M196V, V197M
<400> 13
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Val Met Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 14
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 V197E
<400> 14
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Glu Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 15
<211> 187
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 GGGGS插入在G198和P199之间
<400> 15
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Gly Gly Gly Gly Ser
165 170 175
Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
180 185
<210> 16
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198E
<400> 16
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Glu Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 17
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198D
<400> 17
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Asp Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 18
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198R
<400> 18
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Arg Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 19
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198K
<400> 19
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Lys Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 20
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198Y
<400> 20
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 21
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P199G
<400> 21
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Gly Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 22
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P199R
<400> 22
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Arg Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 23
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P199T
<400> 23
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Thr Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 24
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198P, P199G
<400> 24
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Pro Gly Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 25
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S200Q, Q201S
<400> 25
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Gln Ser Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 26
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 V197D, G198V, P199D, S200H
<400> 26
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Asp Val Asp His Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 27
<211> 185
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 V197-R203缺失并取代为GHRSHLQTVF
<400> 27
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Gly His Arg Ser His Leu Gln
165 170 175
Thr Val Phe Ser Pro Ser Tyr Ala Ser
180 185
<210> 28
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 V197-R203缺失并取代为GLNSMV
<400> 28
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Gly Leu Asn Ser Met Val Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 29
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198缺失
<400> 29
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 30
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198-P199缺失
<400> 30
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 31
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198缺失, P199F
<400> 31
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Phe Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 32
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198缺失, P199L
<400> 32
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Leu Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 33
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198缺失, P199L, S200N
<400> 33
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Leu Asn Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 34
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 V197缺失
<400> 34
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 35
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 L146C, A162C
<400> 35
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Cys Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Cys Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 36
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 A59C, G71C, P199缺失
<400> 36
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Cys
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Cys Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 37
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 A59C, G71C, G198Y
<400> 37
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Cys
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Cys Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 38
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S76C, S79C, P199缺失
<400> 38
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Cys Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 39
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S76C, S79C, G198Y
<400> 39
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Cys Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 40
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G108C, L142C, P199缺失
<400> 40
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Cys Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Cys Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 41
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G108C, L142C, G198Y
<400> 41
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Cys Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Cys Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 42
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D107C, D155C, P199缺失
<400> 42
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Cys
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Cys
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 43
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D107C, D155C, G198Y
<400> 43
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Cys
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Cys
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 44
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55C, P147C, P199缺失
<400> 44
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Cys Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 45
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55C, P147C, G198Y
<400> 45
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Cys Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 46
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55C, N149C, P199缺失
<400> 46
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Cys Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 47
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55C, N149C, G198Y
<400> 47
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Cys Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 48
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 M196缺失
<400> 48
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 49
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S195缺失
<400> 49
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 50
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 L194缺失
<400> 50
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 51
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P193缺失
<400> 51
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 52
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D192缺失
<400> 52
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 53
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S191缺失
<400> 53
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 54
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S190-S191缺失
<400> 54
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 55
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G189缺失
<400> 55
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 56
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 V188缺失
<400> 56
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 57
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D187缺失
<400> 57
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 58
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P186缺失
<400> 58
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 59
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P185-P186缺失
<400> 59
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Asp Val Gly
145 150 155 160
Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 60
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q184缺失
<400> 60
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 61
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S200缺失
<400> 61
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 62
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q201缺失
<400> 62
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 63
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G202缺失
<400> 63
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 64
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R203缺失
<400> 64
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 65
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S204缺失
<400> 65
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 66
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P205缺失
<400> 66
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Ser Tyr Ala Ser
180
<210> 67
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S206缺失
<400> 67
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Tyr Ala Ser
180
<210> 68
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Y207缺失
<400> 68
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Ala Ser
180
<210> 69
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 V197缺失, S201缺失
<400> 69
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Gly Pro Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 70
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 M196缺失, S201缺失
<400> 70
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Val Gly Pro Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 71
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 M196-V197缺失
<400> 71
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 72
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 V197-G198缺失
<400> 72
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 73
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S195-M196缺失
<400> 73
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 74
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G198缺失, S200缺失
<400> 74
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Pro Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 75
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 H60C, S79C
<400> 75
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
Cys Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 76
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 H145C, L165C
<400> 76
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu Cys Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Cys Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 77
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 A59C, G71C
<400> 77
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Cys
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Cys Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 78
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 H60C, A72C
<400> 78
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
Cys Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Cys Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 79
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 I63C, G112C
<400> 79
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Cys Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Cys Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 80
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 L49C, L170C
<400> 80
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Cys Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Cys Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 81
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 T51C, L167C
<400> 81
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Cys Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Cys Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 82
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 H140C, L142C
<400> 82
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
Cys Gly Cys Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 83
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D130C, Y132C
<400> 83
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Cys Gly Cys Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 84
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S137C, H140C
<400> 84
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Cys Glu Ala
100 105 110
Cys Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 85
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S76C, S79C
<400> 85
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Cys Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 86
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G108C, L142C
<400> 86
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Cys Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Cys Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 87
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 A72C, S79C
<400> 87
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Cys Ala Asp Gln
35 40 45
Ser Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 88
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 H60C, S79C, H145C, L165C
<400> 88
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
Cys Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu Cys Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Cys Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 89
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 A72C, S79C, G108C, L142C
<400> 89
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Cys Ala Asp Gln
35 40 45
Ser Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Cys Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Cys Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 90
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R47C, P174C
<400> 90
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Cys Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Cys Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 91
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 M196C, R203C
<400> 91
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Cys Val Gly Pro Ser Gln Gly Cys
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 92
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D107C, D155C
<400> 92
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Cys
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Cys
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 93
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55C, G148C
<400> 93
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Cys Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 94
<211> 175
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55C, G148C, N149-R159缺失并取代为GSGS
<400> 94
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Cys Gly Ser Gly Ser Gly Pro Ala
115 120 125
Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro
130 135 140
Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu
145 150 155 160
Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170 175
<210> 95
<211> 177
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55C, G148C, N149-R159缺失并取代为GGSGGS
<400> 95
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Cys Gly Gly Ser Gly Gly Ser Gly
115 120 125
Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu
130 135 140
Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp
145 150 155 160
Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala
165 170 175
Ser
<210> 96
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55G, Q56C, L146C
<400> 96
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gly Cys Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Cys Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 97
<211> 173
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55G, Q56C, L146C, P147-R159缺失并取代为GSGS
<400> 97
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gly Cys Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Cys Gly Ser Gly Ser Gly Pro Ala Arg Phe
115 120 125
Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro Gly Ile
130 135 140
Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu Ser Met
145 150 155 160
Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170
<210> 98
<211> 175
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55G, Q56C, L146C, P147-R159缺失并取代为
GGSGGS
<400> 98
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gly Cys Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Cys Gly Gly Ser Gly Gly Ser Gly Pro Ala
115 120 125
Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro
130 135 140
Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu
145 150 155 160
Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170 175
<210> 99
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55C, P147C
<400> 99
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Cys Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 100
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q55C, N149C
<400> 100
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Cys Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 101
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 E62C, E78C
<400> 101
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Cys Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Cys Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 102
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 E62C, S79C
<400> 102
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Cys Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 103
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R100C, L114C
<400> 103
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Cys Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Cys His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 104
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 L102C, G112C
<400> 104
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Cys Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Cys Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 105
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 A59C, A73C
<400> 105
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Cys
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Cys Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 106
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 E62C, A72C
<400> 106
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Cys Ile Arg Glu Asp Gly Thr Val Gly Gly Cys Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 107
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q104C, S137C
<400> 107
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Cys Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Cys Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 108
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q104C, H140C
<400> 108
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Cys Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
Cys Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 109
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 A54C, P161C
<400> 109
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Cys Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Cys Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 110
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q56C, L146C
<400> 110
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Cys His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 111
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 I63C, T98C
<400> 111
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Cys Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Cys Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 112
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G67C, R100C
<400> 112
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Cys Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Cys Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 113
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 V69C, L110C
<400> 113
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Cys Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Cys Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 114
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 A120C, A139C
<400> 114
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Cys Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Cys
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 115
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R124C, Q136C
<400> 115
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Cys Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Cys Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 116
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P143C, A162C
<400> 116
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Cys Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Cys Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 117
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P77C, K97C
<400> 117
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Cys Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Cys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 118
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 E78C, T98C
<400> 118
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Cys Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Cys Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 119
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 L80C, V96C
<400> 119
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Cys Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Cys Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 120
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 H145C, P161C
<400> 120
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu Cys Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Cys Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 121
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 L81C, G95C
<400> 121
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Cys Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Cys Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 122
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 Q82C, L94C
<400> 122
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Cys Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Cys Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 123
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 L83C, I91C
<400> 123
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Cys Lys Ala Leu Lys Pro Gly Val Cys
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 124
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D155L
<400> 124
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Leu
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 125
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D155K
<400> 125
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Lys
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 126
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D155Y
<400> 126
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Tyr
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 127
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D155P
<400> 127
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Pro
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 128
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D155E
<400> 128
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Glu
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 129
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 D155N
<400> 129
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asn
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 130
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R154Q, D155L
<400> 130
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Gln Leu
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 131
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R154Q, D155K
<400> 131
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Gln Lys
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 132
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 H153Q, R154Q, D155N, A157V, R159K
<400> 132
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro Gln Gln Asn
115 120 125
Pro Val Pro Lys Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 133
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P152A, H153K, R154K, D155E, P156A, A157S, R159Q
<400> 133
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Ala Lys Lys Glu
115 120 125
Ala Ser Pro Gln Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 134
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 H153Y, R154K, D155N, P156K, A157G
<400> 134
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro Tyr Lys Asn
115 120 125
Lys Gly Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 135
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P152-H153缺失
<400> 135
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Arg Asp Pro Ala
115 120 125
Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala
130 135 140
Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly
145 150 155 160
Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 136
<211> 171
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 N149-R163缺失并取代为GSGS
<400> 136
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Gly Ser Gly Ser Phe Leu Pro
115 120 125
Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala
130 135 140
Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly
145 150 155 160
Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170
<210> 137
<211> 173
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 N149-A162缺失并取代为GSHSG
<400> 137
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Gly Ser His Ser Gly Arg Phe
115 120 125
Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro Gly Ile
130 135 140
Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu Ser Met
145 150 155 160
Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170
<210> 138
<211> 174
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 N149-A162缺失并取代为GSHSGS
<400> 138
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Gly Ser His Ser Gly Ser Arg
115 120 125
Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro Gly
130 135 140
Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu Ser
145 150 155 160
Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170
<210> 139
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 K150H
<400> 139
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn His Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 140
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 K150H, P152L
<400> 140
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn His Ser Leu His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 141
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R163H
<400> 141
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala His Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 142
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P158H, R159H
<400> 142
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala His His Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 143
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 N149-D155缺失并取代为GSGS
<400> 143
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Gly Ser Gly Ser Pro Ala Pro
115 120 125
Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro
130 135 140
Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser
145 150 155 160
Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 144
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 N149-D155缺失并取代为GSHSG
<400> 144
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Gly Ser His Ser Gly Pro Ala
115 120 125
Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala
130 135 140
Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly
145 150 155 160
Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 145
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 N149-D155缺失并取代为ATTS
<400> 145
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Ala Thr Thr Ser Pro Ala Pro
115 120 125
Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro
130 135 140
Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser
145 150 155 160
Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 146
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R159-R163缺失并取代为GA
<400> 146
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Gly Ala Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro
130 135 140
Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser
145 150 155 160
Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 147
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R159-R163缺失并取代为GY
<400> 147
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Gly Tyr Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro
130 135 140
Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser
145 150 155 160
Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 148
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R159-R163缺失并取代为HH
<400> 148
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro His His Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro
130 135 140
Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser
145 150 155 160
Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 149
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R159-R163缺失并取代为GE
<400> 149
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Gly Glu Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro
130 135 140
Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser
145 150 155 160
Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 150
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R159-R163缺失并取代为HE
<400> 150
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro His Glu Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro
130 135 140
Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser
145 150 155 160
Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 151
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 A162Y
<400> 151
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Tyr Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 152
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R159H
<400> 152
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro His Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 153
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 S200M
<400> 153
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Met Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 154
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G202T
<400> 154
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Thr Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 155
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R203E
<400> 155
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Glu
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 156
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 R203H
<400> 156
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly His
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 157
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 G202-R203缺失
<400> 157
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 158
<211> 143
<212> PRT
<213> 人工序列
<220>
<223> FGF21 (S34-E176), Q55C, N149C
<400> 158
Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val Arg Gln Arg Tyr Leu
1 5 10 15
Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala His Leu Glu Ile Arg Glu
20 25 30
Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser Pro Glu Ser Leu Leu
35 40 45
Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln Ile Leu Gly Val Lys
50 55 60
Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly Ala Leu Tyr Gly Ser
65 70 75 80
Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg Glu Leu Leu Leu Glu
85 90 95
Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His Gly Leu Pro Leu His
100 105 110
Leu Pro Gly Cys Lys Ser Pro His Arg Asp Pro Ala Pro Arg Gly Pro
115 120 125
Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu
130 135 140
<210> 159
<211> 199
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 PLSMVGPSQGRSPSYAS插入在S209之后
<400> 159
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser Pro Leu Ser Met Val Gly Pro Ser Gln Gly
180 185 190
Arg Ser Pro Ser Tyr Ala Ser
195
<210> 160
<211> 198
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 P199缺失, PLSMVGSQGRSPSYAS插入在S209之后
<400> 160
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser Pro Leu Ser Met Val Gly Ser Gln Gly Arg
180 185 190
Ser Pro Ser Tyr Ala Ser
195
<210> 161
<211> 4
<212> PRT
<213> 人工序列
<220>
<223> 接头
<400> 161
Gly Ser Gly Ser
1
<210> 162
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 接头
<400> 162
Gly Gly Ser Gly Gly Ser
1 5
<210> 163
<211> 5
<212> PRT
<213> 人工序列
<220>
<223> 接头
<400> 163
Gly Ser His Ser Gly
1 5
<210> 164
<211> 4
<212> PRT
<213> 人工序列
<220>
<223> 接头
<400> 164
Ala Thr Thr Ser
1
<210> 165
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 接头
<400> 165
Gly Ser His Ser Gly Ser
1 5
<210> 166
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> 接头
<400> 166
Gly His Arg Ser His Leu Gln Thr Val Phe
1 5 10
<210> 167
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 接头
<400> 167
Gly Leu Asn Ser Met Val
1 5
<210> 168
<211> 5
<212> PRT
<213> 人工序列
<220>
<223> 接头
<400> 168
Gly Gly Gly Gly Ser
1 5
<210> 169
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> C-末端延伸
<400> 169
Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala
1 5 10 15
Ser
<210> 170
<211> 16
<212> PRT
<213> 人工序列
<220>
<223> C-末端延伸
<400> 170
Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
1 5 10 15
<210> 171
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21变体, 共有序列I
<220>
<221> misc_feature
<222> (27)..(27)
<223> Xaa (= Xaa55)为Q或另一氨基酸
<220>
<221> misc_feature
<222> (119)..(119)
<223> Xaa (= Xaa147)为P或C或缺失
<220>
<221> misc_feature
<222> (120)..(120)
<223> Xaa (= Xaa148)为G或C或缺失
<220>
<221> misc_feature
<222> (121)..(121)
<223> Xaa (= Xaa149)为N或C或缺失
<220>
<221> misc_feature
<222> (122)..(122)
<223> Xaa (= Xaa150)为K或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (123)..(123)
<223> Xaa (= Xaa151)为S或缺失
<220>
<221> misc_feature
<222> (124)..(124)
<223> Xaa (= Xaa152)为P或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (125)..(125)
<223> Xaa (= Xaa153)为H或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (126)..(126)
<223> Xaa (= Xaa154)为R或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (127)..(127)
<223> Xaa (= Xaa155)为D或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (128)..(128)
<223> Xaa (= Xaa156)为P或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (129)..(129)
<223> Xaa (= Xaa157)为A或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (130)..(130)
<223> Xaa (= Xaa158)为P或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (131)..(131)
<223> Xaa (= Xaa159)为R或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (132)..(132)
<223> Xaa (= Xaa160)为G或缺失
<220>
<221> misc_feature
<222> (133)..(133)
<223> Xaa (= Xaa161)为P或C或缺失
<220>
<221> misc_feature
<222> (134)..(134)
<223> Xaa (= Xaa162)为A或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (135)..(135)
<223> Xaa (= Xaa163)为R或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (156)..(156)
<223> Xaa (= Xaa184)为Q或缺失
<220>
<221> misc_feature
<222> (157)..(157)
<223> Xaa (= Xaa185)为P或缺失
<220>
<221> misc_feature
<222> (158)..(158)
<223> Xaa (= Xaa186)为P或缺失
<220>
<221> misc_feature
<222> (159)..(159)
<223> Xaa (= Xaa187)为D或缺失
<220>
<221> misc_feature
<222> (160)..(160)
<223> Xaa (= Xaa188)为V或缺失
<220>
<221> misc_feature
<222> (161)..(161)
<223> Xaa (= Xaa189)为G或缺失
<220>
<221> misc_feature
<222> (162)..(162)
<223> Xaa (= Xaa190)为S或缺失
<220>
<221> misc_feature
<222> (163)..(163)
<223> Xaa (= Xaa191)为S或缺失
<220>
<221> misc_feature
<222> (164)..(164)
<223> Xaa (= Xaa192)为D或缺失
<220>
<221> misc_feature
<222> (165)..(165)
<223> Xaa (= Xaa193)为P或缺失
<220>
<221> misc_feature
<222> (166)..(166)
<223> Xaa (= Xaa194)为L或缺失
<220>
<221> misc_feature
<222> (167)..(167)
<223> Xaa (= Xaa195)为S或C或缺失
<220>
<221> misc_feature
<222> (168)..(168)
<223> Xaa (= Xaa196)为M或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (169)..(169)
<223> Xaa (= Xaa197)为V或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (170)..(170)
<223> Xaa (= Xaa198)为G或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (171)..(171)
<223> Xaa (= Xaa199)为P或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (172)..(172)
<223> Xaa (= Xaa200)为S或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (173)..(173)
<223> Xaa (= Xaa201)为Q或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (174)..(174)
<223> Xaa (= Xaa202)为G或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (175)..(175)
<223> Xaa (= Xaa203)为R或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (176)..(176)
<223> Xaa (= Xaa204)为S或缺失
<220>
<221> misc_feature
<222> (177)..(177)
<223> Xaa (= Xaa205)为P或缺失
<220>
<221> misc_feature
<222> (178)..(178)
<223> Xaa (= Xaa206)为S或缺失
<220>
<221> misc_feature
<222> (179)..(179)
<223> Xaa (= Xaa207)为Y或缺失
<400> 171
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Xaa Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
115 120 125
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Xaa Xaa Xaa Xaa Xaa
145 150 155 160
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
165 170 175
Xaa Xaa Xaa Ala Ser
180
<210> 172
<211> 209
<212> PRT
<213> 人(Homo sapiens)
<400> 172
Met Asp Ser Asp Glu Thr Gly Phe Glu His Ser Gly Leu Trp Val Ser
1 5 10 15
Val Leu Ala Gly Leu Leu Leu Gly Ala Cys Gln Ala His Pro Ile Pro
20 25 30
Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val Arg Gln Arg Tyr
35 40 45
Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His Leu Glu Ile Arg
50 55 60
Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser Pro Glu Ser Leu
65 70 75 80
Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln Ile Leu Gly Val
85 90 95
Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly Ala Leu Tyr Gly
100 105 110
Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg Glu Leu Leu Leu
115 120 125
Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His Ser Leu Pro Leu
130 135 140
His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro Ala Pro Arg Gly
145 150 155 160
Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu
165 170 175
Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp
180 185 190
Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala
195 200 205
Ser
<210> 173
<211> 209
<212> PRT
<213> 人(Homo sapiens)
<400> 173
Met Asp Ser Asp Glu Thr Gly Phe Glu His Ser Gly Leu Trp Val Ser
1 5 10 15
Val Leu Ala Gly Leu Leu Leu Gly Ala Cys Gln Ala His Pro Ile Pro
20 25 30
Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val Arg Gln Arg Tyr
35 40 45
Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His Leu Glu Ile Arg
50 55 60
Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser Pro Glu Ser Leu
65 70 75 80
Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln Ile Leu Gly Val
85 90 95
Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly Ala Leu Tyr Gly
100 105 110
Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg Glu Leu Leu Leu
115 120 125
Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His Gly Leu Pro Leu
130 135 140
His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro Ala Pro Arg Gly
145 150 155 160
Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Leu Pro Glu
165 170 175
Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp
180 185 190
Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala
195 200 205
Ser
<210> 174
<211> 182
<212> PRT
<213> 人工序列
<220>
<223> G-FGF21 (L174变体)
<400> 174
Gly His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln
1 5 10 15
Val Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala
20 25 30
His Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln
35 40 45
Ser Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile
50 55 60
Gln Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp
65 70 75 80
Gly Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe
85 90 95
Arg Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala
100 105 110
His Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp
115 120 125
Pro Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro
130 135 140
Pro Ala Leu Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp
145 150 155 160
Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
165 170 175
Ser Pro Ser Tyr Ala Ser
180
<210> 175
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21变体, 共有序列II
<220>
<221> misc_feature
<222> (27)..(27)
<223> Xaa (= Xaa55)为Q或C
<220>
<221> misc_feature
<222> (119)..(119)
<223> Xaa (= Xaa147)为P或C
<220>
<221> misc_feature
<222> (120)..(120)
<223> Xaa (= Xaa148)为G或C
<220>
<221> misc_feature
<222> (121)..(121)
<223> Xaa (= Xaa149)为N或C
<220>
<221> misc_feature
<222> (127)..(127)
<223> Xaa (= Xaa155)为D或C
<220>
<221> misc_feature
<222> (133)..(133)
<223> Xaa (= Xaa161)为P或C
<220>
<221> misc_feature
<222> (134)..(134)
<223> Xaa (= Xaa162)为A或C
<220>
<221> misc_feature
<222> (167)..(167)
<223> Xaa (= Xaa195)为S或C
<220>
<221> misc_feature
<222> (168)..(168)
<223> Xaa (= Xaa196)为M或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (169)..(169)
<223> Xaa (= Xaa197)为V或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (170)..(170)
<223> Xaa (= Xaa198)为G或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (171)..(171)
<223> Xaa (= Xaa199)为P或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (172)..(172)
<223> Xaa (= Xaa200)为S或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (173)..(173)
<223> Xaa (= Xaa201)为Q或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (174)..(174)
<223> Xaa (= Xaa202)为G或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (175)..(175)
<223> Xaa (= Xaa203)为R或另一氨基酸或缺失
<400> 175
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Xaa Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Xaa Xaa Xaa Lys Ser Pro His Arg Xaa Pro
115 120 125
Ala Pro Arg Gly Xaa Xaa Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 176
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21变体, 共有序列III
<220>
<221> misc_feature
<222> (27)..(27)
<223> Xaa (= Xaa55)为Q或C
<220>
<221> misc_feature
<222> (119)..(119)
<223> Xaa (= Xaa147)为P或C
<220>
<221> misc_feature
<222> (121)..(121)
<223> Xaa (= Xaa149)为N或C
<220>
<221> misc_feature
<222> (127)..(127)
<223> Xaa (= Xaa155)为D或C
<220>
<221> misc_feature
<222> (170)..(170)
<223> Xaa (= Xaa198)为G或另一氨基酸或缺失
<220>
<221> misc_feature
<222> (171)..(171)
<223> Xaa (= Xaa199)为P或另一氨基酸或缺失
<400> 2
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Xaa Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Xaa Gly Xaa Lys Ser Pro His Arg Xaa Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Xaa Xaa Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 177
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S195C, S200C
<400> 177
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Cys Met Val Gly Pro Cys Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 178
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P199S, S200P
<400> 178
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Pro Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 179
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P199Q, Q201P
<400> 179
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Gln Ser Pro Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 180
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P199缺失
<400> 180
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 181
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 M196P, V197G, G198V, P199M
<400> 181
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Pro Gly Val Met Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 182
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 M196V, V197M
<400> 182
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Val Met Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 183
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 V197E
<400> 183
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Glu Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 184
<211> 186
<212> PRT
<213> 人工序列
<220>
<223> FGF21 GGGGS (SEQ ID NO: 168)插入在G198和P199之间
<400> 184
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Gly Gly Gly Gly Ser Pro
165 170 175
Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
180 185
<210> 185
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198E
<400> 185
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Glu Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 186
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198D
<400> 186
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Asp Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 187
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198R
<400> 187
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Arg Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 188
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198K
<400> 188
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Lys Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 189
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198Y
<400> 189
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 190
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P199G
<400> 190
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Gly Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 191
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P199R
<400> 191
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Arg Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 192
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P199T
<400> 192
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Thr Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 193
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198P, P199G
<400> 193
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Pro Gly Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 194
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S200Q, Q201S
<400> 194
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Gln Ser Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 195
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 V197D, G198V, P199D, S200H
<400> 195
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Asp Val Asp His Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 196
<211> 184
<212> PRT
<213> 人工序列
<220>
<223> FGF21 V197-R203缺失并取代为GHRSHLQTVF (SEQ ID NO:
166)
<400> 196
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Gly His Arg Ser His Leu Gln Thr
165 170 175
Val Phe Ser Pro Ser Tyr Ala Ser
180
<210> 197
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 V197-R203缺失并取代为GLNSMV (SEQ ID NO: 167)
<400> 197
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Gly Leu Asn Ser Met Val Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 198
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198缺失
<400> 198
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 199
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198-P199缺失
<400> 199
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 200
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198缺失, P199F
<400> 200
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Phe Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 201
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198缺失, P199L
<400> 201
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Leu Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 202
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198缺失, P199L, S200N
<400> 202
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Leu Asn Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 203
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 V197缺失
<400> 203
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 204
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 L146C, A162C
<400> 204
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Cys Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Cys Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 205
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 A59C, G71C, P199缺失
<400> 205
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Cys His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Cys Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 206
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 A59C, G71C, G198Y
<400> 206
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Cys His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Cys Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 207
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S76C, S79C, P199缺失
<400> 207
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Cys
35 40 45
Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 208
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S76C, S79C, G198Y
<400> 208
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Cys
35 40 45
Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 209
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G108C, L142C, P199缺失
<400> 209
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Cys
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Cys Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 210
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G108C, L142C, G198Y
<400> 210
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Cys
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Cys Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 211
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D107C, D155C, P199缺失
<400> 211
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Cys Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Cys Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 212
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D107C, D155C, G198Y
<400> 212
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Cys Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Cys Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 213
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55C, P147C, P199缺失
<400> 213
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Cys Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 214
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55C, P147C, G198Y
<400> 214
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Cys Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 215
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55C, N149C, P199缺失
<400> 215
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Cys Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 216
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55C, N149C, G198Y
<400> 216
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Cys Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Tyr Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 217
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 M196缺失
<400> 217
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 218
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S195缺失
<400> 218
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 219
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 L194缺失
<400> 219
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 220
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P193缺失
<400> 220
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 221
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D192缺失
<400> 221
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 222
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S191缺失
<400> 222
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 223
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S190-S191缺失
<400> 223
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 224
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G189缺失
<400> 224
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 225
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 V188缺失
<400> 225
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Gly
145 150 155 160
Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 226
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D187缺失
<400> 226
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Val Gly
145 150 155 160
Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 227
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P186缺失
<400> 227
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Asp Val Gly
145 150 155 160
Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 228
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P185-P186缺失
<400> 228
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Asp Val Gly Ser
145 150 155 160
Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 229
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q184缺失
<400> 229
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Pro Pro Asp Val Gly
145 150 155 160
Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 230
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S200缺失
<400> 230
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Gln Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 231
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q201缺失
<400> 231
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gly Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 232
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G202缺失
<400> 232
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Arg Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 233
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R203缺失
<400> 233
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Ser Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 234
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S204缺失
<400> 234
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Pro
165 170 175
Ser Tyr Ala Ser
180
<210> 235
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P205缺失
<400> 235
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Ser Tyr Ala Ser
180
<210> 236
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S206缺失
<400> 236
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Tyr Ala Ser
180
<210> 237
<211> 180
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Y207缺失
<400> 237
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Ala Ser
180
<210> 238
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 V197缺失, S201缺失
<400> 238
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Gly Pro Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 239
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 M196缺失, S201缺失
<400> 239
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Val Gly Pro Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 240
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 M196-V197缺失
<400> 240
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 241
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 V197-G198缺失
<400> 241
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 242
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S195-M196缺失
<400> 242
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 243
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G198缺失, S200缺失
<400> 243
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Pro Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 244
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 H60C, S79C
<400> 244
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala Cys
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 245
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 H145C, L165C
<400> 245
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu Cys Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Cys Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 246
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 A59C, G71C
<400> 246
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Cys His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Cys Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 247
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 H60C, A72C
<400> 247
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala Cys
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Cys Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 248
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 I63C, G112C
<400> 248
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Cys Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Cys Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 249
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 L49C, L170C
<400> 249
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Cys Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Cys Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 250
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 T51C, L167C
<400> 250
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Cys Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Cys Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 251
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 H140C, L142C
<400> 251
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala Cys
100 105 110
Gly Cys Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 252
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D130C, Y132C
<400> 252
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Cys Gly Cys Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 253
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S137C, H140C
<400> 253
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Cys Glu Ala Cys
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 254
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S76C, S79C
<400> 254
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Cys
35 40 45
Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 255
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G108C, L142C
<400> 255
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Cys
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Cys Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 256
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 A72C, S79C
<400> 256
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Cys Ala Asp Gln Ser
35 40 45
Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 257
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 H60C, S79C, H145C, L165C
<400> 257
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala Cys
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu Cys Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Cys Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 258
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 A72C, S79C, G108C, L142C
<400> 258
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Cys Ala Asp Gln Ser
35 40 45
Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Cys
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Cys Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 259
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R47C, P174C
<400> 259
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Cys Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Cys Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 260
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 M196C, R203C
<400> 260
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Cys Val Gly Pro Ser Gln Gly Cys Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 261
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D107C, D155C
<400> 261
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Cys Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Cys Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 262
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55C, G148C
<400> 262
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Cys Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 263
<211> 174
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55C, G148C, N149-R159缺失并取代为GSGS (SEQ ID
NO: 161)
<400> 263
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Cys Gly Ser Gly Ser Gly Pro Ala Arg
115 120 125
Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro Gly
130 135 140
Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu Ser
145 150 155 160
Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170
<210> 264
<211> 176
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55C, G148C, N149-R159缺失并取代为GGSGGS (SEQ
ID NO: 162)
<400> 264
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Cys Gly Gly Ser Gly Gly Ser Gly Pro
115 120 125
Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro
130 135 140
Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro
145 150 155 160
Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170 175
<210> 265
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55G, Q56C, L146C
<400> 265
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gly Cys Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Cys Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 266
<211> 172
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55G, Q56C, L146C, P147-R159缺失并取代为GSGS
(SEQ ID NO: 161)
<400> 266
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gly Cys Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Cys Gly Ser Gly Ser Gly Pro Ala Arg Phe Leu
115 120 125
Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu
130 135 140
Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu Ser Met Val
145 150 155 160
Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170
<210> 267
<211> 174
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55G, Q56C, L146C, P147-R159缺失并取代为GGSGGS
(SEQ ID NO: 162)
<400> 267
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gly Cys Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Cys Gly Gly Ser Gly Gly Ser Gly Pro Ala Arg
115 120 125
Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro Gly
130 135 140
Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu Ser
145 150 155 160
Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170
<210> 268
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55C, P147C
<400> 268
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Cys Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 269
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q55C, N149C
<400> 269
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Cys Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 270
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 E62C, E78C
<400> 270
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Cys Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Cys Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 271
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 E62C, S79C
<400> 271
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Cys Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Cys Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 272
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R100C, L114C
<400> 272
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Cys Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Cys His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 273
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 L102C, G112C
<400> 273
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Cys Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Cys Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 274
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 A59C, A73C
<400> 274
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Cys His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Cys Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 275
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 E62C, A72C
<400> 275
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Cys Ile Arg Glu Asp Gly Thr Val Gly Gly Cys Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 276
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q104C, S137C
<400> 276
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Cys Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Cys Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 277
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q104C, H140C
<400> 277
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Cys Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala Cys
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 278
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 A54C, P161C
<400> 278
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Cys Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Cys Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 279
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q56C, L146C
<400> 279
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Cys Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Cys His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 280
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 I63C, T98C
<400> 280
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Cys Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Cys Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 281
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G67C, R100C
<400> 281
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Cys Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Cys Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 282
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 V69C, L110C
<400> 282
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Cys Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Cys Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 283
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 A120C, A139C
<400> 283
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Cys Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Cys His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 284
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R124C, Q136C
<400> 284
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Cys
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Cys Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 285
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P143C, A162C
<400> 285
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Cys Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Cys Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 286
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P77C, K97C
<400> 286
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Cys Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Cys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 287
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 E78C, T98C
<400> 287
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Cys Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Cys Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 288
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 L80C, V96C
<400> 288
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Cys Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Cys Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 289
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 H145C, P161C
<400> 289
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu Cys Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Cys Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 290
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 L81C, G95C
<400> 290
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Cys Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Cys Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 291
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 Q82C, L94C
<400> 291
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Cys Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Cys Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 292
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 L83C, I91C
<400> 292
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Cys Lys Ala Leu Lys Pro Gly Val Cys Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 293
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D155L
<400> 293
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Leu Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 294
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D155K
<400> 294
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Lys Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 295
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D155Y
<400> 295
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Tyr Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 296
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D155P
<400> 296
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Pro Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 297
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D155E
<400> 297
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Glu Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 298
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 D155N
<400> 298
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asn Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 299
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R154Q, D155L
<400> 299
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Gln Leu Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 300
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R154Q, D155K
<400> 300
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Gln Lys Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 301
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 H153Q, R154Q, D155N, A157V, R159K
<400> 301
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro Gln Gln Asn Pro
115 120 125
Val Pro Lys Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 302
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P152A, H153K, R154K, D155E, P156A, A157S, R159Q
<400> 302
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Ala Lys Lys Glu Ala
115 120 125
Ser Pro Gln Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 303
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 H153Y, R154K, D155N, P156K, A157G
<400> 303
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro Tyr Lys Asn Lys
115 120 125
Gly Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 304
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P152-H153缺失
<400> 304
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Arg Asp Pro Ala Pro
115 120 125
Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro
130 135 140
Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser
145 150 155 160
Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 305
<211> 170
<212> PRT
<213> 人工序列
<220>
<223> FGF21 N149-R163缺失并取代为GSGS (SEQ ID NO: 161)
<400> 305
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Gly Ser Gly Ser Phe Leu Pro Leu
115 120 125
Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro
130 135 140
Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro
145 150 155 160
Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170
<210> 306
<211> 172
<212> PRT
<213> 人工序列
<220>
<223> FGF21 N149-A162缺失并取代为GSHSG (SEQ ID NO: 163)
<400> 306
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Gly Ser His Ser Gly Arg Phe Leu
115 120 125
Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro Gly Ile Leu
130 135 140
Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu Ser Met Val
145 150 155 160
Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170
<210> 307
<211> 173
<212> PRT
<213> 人工序列
<220>
<223> FGF21 N149-A162缺失并取代为GSHSGS (SEQ ID NO: 165)
<400> 307
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Gly Ser His Ser Gly Ser Arg Phe
115 120 125
Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro Glu Pro Pro Gly Ile
130 135 140
Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp Pro Leu Ser Met
145 150 155 160
Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala Ser
165 170
<210> 308
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 K150H
<400> 308
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn His Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 309
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 K150H, P152L
<400> 309
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn His Ser Leu His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 310
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R163H
<400> 310
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala His Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 311
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P158H, R159H
<400> 311
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala His His Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 312
<211> 178
<212> PRT
<213> 人工序列
<220>
<223> FGF21 N149-D155缺失并取代为GSGS (SEQ ID NO: 161)
<400> 312
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Gly Ser Gly Ser Pro Ala Pro Arg
115 120 125
Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro
130 135 140
Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser
145 150 155 160
Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr
165 170 175
Ala Ser
<210> 313
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 N149-D155缺失并取代为GSHSG (SEQ ID NO: 163)
<400> 313
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Gly Ser His Ser Gly Pro Ala Pro
115 120 125
Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro
130 135 140
Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser
145 150 155 160
Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 314
<211> 178
<212> PRT
<213> 人工序列
<220>
<223> FGF21 N149-D155缺失并取代为ATTS (SEQ ID NO: 164)
<400> 314
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Ala Thr Thr Ser Pro Ala Pro Arg
115 120 125
Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro
130 135 140
Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser
145 150 155 160
Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr
165 170 175
Ala Ser
<210> 315
<211> 178
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R159-R163缺失并取代为GA
<400> 315
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Gly Ala Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro
130 135 140
Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser
145 150 155 160
Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr
165 170 175
Ala Ser
<210> 316
<211> 178
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R159-R163缺失并取代为GY
<400> 316
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Gly Tyr Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro
130 135 140
Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser
145 150 155 160
Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr
165 170 175
Ala Ser
<210> 317
<211> 178
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R159-R163缺失并取代为HH
<400> 317
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro His His Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro
130 135 140
Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser
145 150 155 160
Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr
165 170 175
Ala Ser
<210> 318
<211> 178
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R159-R163缺失并取代为GE
<400> 318
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Gly Glu Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro
130 135 140
Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser
145 150 155 160
Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr
165 170 175
Ala Ser
<210> 319
<211> 178
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R159-R163缺失并取代为HE
<400> 319
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro His Glu Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Pro Pro
130 135 140
Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser
145 150 155 160
Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr
165 170 175
Ala Ser
<210> 320
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 A162Y
<400> 320
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Tyr Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 321
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R159H
<400> 321
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro His Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 322
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 S200M
<400> 322
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Met Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 323
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G202T
<400> 323
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Thr Arg Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 324
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R203E
<400> 324
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Glu Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 325
<211> 181
<212> PRT
<213> 人工序列
<220>
<223> FGF21 R203H
<400> 325
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly His Ser
165 170 175
Pro Ser Tyr Ala Ser
180
<210> 326
<211> 179
<212> PRT
<213> 人工序列
<220>
<223> FGF21 G202-R203缺失
<400> 326
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Ser Pro Ser
165 170 175
Tyr Ala Ser
<210> 327
<211> 198
<212> PRT
<213> 人工序列
<220>
<223> FGF21 PLSMVGPSQGRSPSYAS (SEQ ID NO: 169)插入在S209之后
<400> 327
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg
180 185 190
Ser Pro Ser Tyr Ala Ser
195
<210> 328
<211> 197
<212> PRT
<213> 人工序列
<220>
<223> FGF21 P199缺失, PLSMVGSQGRSPSYAS (SEQ ID NO: 170) inserted
after S209
<400> 328
His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val
1 5 10 15
Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His
20 25 30
Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser
35 40 45
Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln
50 55 60
Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly
65 70 75 80
Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg
85 90 95
Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His
100 105 110
Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro
115 120 125
Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro
130 135 140
Ala Pro Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val
145 150 155 160
Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser
165 170 175
Pro Ser Tyr Ala Ser Pro Leu Ser Met Val Gly Ser Gln Gly Arg Ser
180 185 190
Pro Ser Tyr Ala Ser
195

Claims (32)

1.一种人成纤维细胞生长因子21(FGF21)的变体，其包含SEQ ID NO:171的氨基酸序列，
其中
-Xaa55是Q或C；
-Xaa147是P或C；
-Xaa148是G或C；
-Xaa149是N或C；
-Xaa150是K；
-Xaa151是S；
-Xaa152是P；
-Xaa153是H；
-Xaa154是R；
-Xaa155是D或C；
-Xaa156是P；
-Xaa157是A；
-Xaa158是P；
-Xaa159是R；
-Xaa160是G；
-Xaa161是P或C；
-Xaa162是A或C；
-Xaa163是R；
-Xaa184是Q；
-Xaa185是P；
-Xaa186是P；
-Xaa187是D；
-Xaa188是V；
-Xaa189是G；
-Xaa190是S；
-Xaa191是S；
-Xaa192是D；
-Xaa193是P；
-Xaa194是L；
-Xaa195是S或C；
-Xaa196是M或另一氨基酸，如P，V或C，或是缺失的；
-Xaa197是V或另一氨基酸，如E，D，G或M，或是缺失的；
-Xaa198是G或另一氨基酸，如E，D，R，K，Y，P或V，或是缺失的；
-Xaa199是P或另一氨基酸，如S，Q，R，T，G，F，L，D或M，或是缺失的；
-Xaa200是S或另一氨基酸，如Q，M，C，P，N或H，或是缺失的；
-Xaa201是Q或另一氨基酸，如P或S，或是缺失的；
-Xaa202是G或另一氨基酸，如T，或是缺失的；
-Xaa203是R或另一氨基酸，如E，H或C，或是缺失的；
-Xaa204是S；
-Xaa205是P；
-Xaa206是S；且
-Xaa207是Y；
其中，任选地，SEQ ID NO:171包含以C对如下氨基酸中至少一个的取代：R47，L49，T51，A54，Q56，A59，H60，E62，I63，G67，V69，G71，A72，A73，S76，P77，E78，S79，L80，L81，Q82，L83，I91，L94，G95，V96，K97，T98，R100，L102，Q104，D107，G108，L110，G112，L114，A120，R124，D130，Y132，Q136，S137，A139，H140，L142，P143，H145，L146，L165，L167，L170，P174；
其条件是SEQ ID NO:171不是成熟人野生型FGF21(SEQ ID NO:2)且与成熟人野生型FGF21(SEQ ID NO:2)相比包含0，2，4，6或8个额外的半胱氨酸，
其中，任选地，SEQ ID NO:171还包含突变G141S和/或突变P174L。
2.如权利要求1的变体，其包含SEQ ID NO:175的氨基酸序列。
3.如权利要求1或2的变体，其中
-
-Xaa197至Xaa203被缺失并由蛋白酶抗性肽接头替代，如选自SEQ ID NO:166和SEQ IDNO:167；
-蛋白酶抗性肽接头，如SEQ ID NO:168，被插入Xaa198和Xaa199之间；和/或
-SEQ ID NO:169或SEQ ID NO:170的氨基酸序列被添加于SEQ ID NO:171的S209之后。
4.如权利要求1-3任一项的变体，其中SEQ ID NO:171与成熟人野生型FGF21(SEQ IDNO:2)具有至少90％或至少91％或至少92％或至少93％或至少94％或至少95％或至少96％或至少97％或至少98％序列同一性。
5.如权利要求1-4任一项的变体，其中
-Xaa55是Q或C；
-Xaa147是P或C；
-Xaa148是G；
-Xaa149是N或C；
-Xaa155是D或C；
-Xaa161是P；
-Xaa162是A；
-Xaa195是S；
-Xaa196是M；
-Xaa197是V；
-Xaa198是G或另一氨基酸，如E，D，R，K，Y，P或V，或是缺失的；
-Xaa199是P或另一氨基酸，如S，Q，R，T，G，F，L，D或M，或是缺失的；
-Xaa200是S；
-Xaa201是Q；
-Xaa202是G；且
-Xaa203是R；
其中，任选地，SEQ ID NO:171包含以C对如下氨基酸中至少一个的取代：R47，A59，H60，G71，S76，S79，D107，G108，L142，P174。
6.如权利要求5的变体，其包含SEQ ID NO:176的氨基酸序列。
7.如权利要求1-6任一项的变体，其中SEQ ID NO:171包含以C对如下氨基酸的取代：
-Xaa55和Xaa149；
-R47和P174；
-Xaa55和Xaa147；
-A59和G71；
-H60和S79；
-S76和S79；
-D107和Xaa155；和/或
-G108和L142。
8.如权利要求1-7任一项的变体，其中
-Xaa55是C，Xaa147是C，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是缺失的；
-Xaa55是C，Xaa147是C，Xaa149是N，Xaa155是D，Xaa198是Y，且Xaa199是P；
-Xaa55是C，Xaa147是P，Xaa149是C，Xaa155是D，Xaa198是G，且Xaa199是缺失的；
-Xaa55是C，Xaa147是P，Xaa149是C，Xaa155是D，Xaa198是Y，且Xaa199是P；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是缺失的；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是R，且Xaa199是P；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是K，且Xaa199是P；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是Y，且Xaa199是P；
-Xaa55是C，Xaa147是C，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是P；
-Xaa55是C，Xaa147是P，Xaa149是C，Xaa155是D，Xaa198是G，且Xaa199是P；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是缺失的，其中SEQ ID NO:171包含以C对A59和G71的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是Y，且Xaa199是P，其中SEQ IDNO:171包含以C对A59和G71的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是缺失的，其中SEQ ID NO:171包含以C对S76和S79的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是Y，且Xaa199是P，其中SEQ IDNO:171包含以C对S76和S79的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是G，且Xaa199是缺失的，其中SEQ ID NO:171包含以C对G108和L142的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是D，Xaa198是Y，且Xaa199是P，其中SEQ IDNO:171包含以C对G108和L142的取代；
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是C，Xaa198是G，且Xaa199是缺失的，其中SEQ ID NO:171包含以C对D107的取代；或
-Xaa55是Q，Xaa147是P，Xaa149是N，Xaa155是C，Xaa198是Y，且Xaa199是P，其中SEQ IDNO:171包含以C对D107的取代。
9.如权利要求1-8任一项的变体，其在其N末端进一步包含至少一个额外氨基酸。
10.如权利要求9的变体，其中该至少一个额外氨基酸选自由下列组成的组：G、A、N和C。
11.一种人FGF21的变体，其包含根据SEQ ID NO:269、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、61、62、63、64、69、70、71、72、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、89、90、91、92、93、99、100、101、102、103、104、105、106、107、108、109、110、111、112、113、114、115、116、117、118、119、120、121、122、123、153、154、155、156、157、158、159 160、177、178、179、180、181、182、183、184、185、186、187、188、189、190、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、230、231、232、233、238、239、240、241、243、244、245、246、247、248、249、250、251、252、253、254、255、256、257、258、259、260、261、262、268、270、271、272、273、274、275、276、277、278、279、280、281、282、283、284、285、286、287、288、289、290、291、292、322、323、324、325、326、327和328之一的氨基酸序列或由其组成，其中，任选地，该变体进一步包含突变G141S和/或突变P174L，其中该氨基酸的编号依照SEQ ID NO:1。
12.如权利要求1-11任一项的变体，其中该变体与成熟人野生型FGF21(SEQ ID NO:2)相比在人和/或鼠血浆中具有增加的蛋白水解稳定性。
13.如权利要求1-12任一项的变体，其中该变体与成熟人野生型FGF21(SEQ ID NO:2)相比具有增加的热稳定性。
14.如权利要求1-13任一项的变体，其中该变体诱导促分裂原活化蛋白激酶(MAPK)ERK1/2的磷酸化。
15.如权利要求14的变体，其中该变体以100nmol/L或更低、或90nmol/L或更低、或80nmol/L或更低、或70nmol/L或更低、或60nmol/L或更低、或50nmol/L或更低、或40nmol/L或更低、或30nmol/L或更低、或20nmol/L或更低、或15nmol/L或更低、或12nmol/L或更低、或11nmol/L或更低、或10nmol/L或更低、或9nmol/L或更低、或8nmol/L或更低、或7nmol/L或更低、或6nmol/L或更低、或5nmol/L或更低、或4nmol/L或更低、或3nmol/L或更低、或2nmol/L或更低的EC50诱导促分裂原活化蛋白激酶(MAPK)ERK1/2的磷酸化，该EC50如在In-CellWestern(ICW)测定法中所确定的。
16.如权利要求1-15任一项的变体，其还包含至少一个允许对该变体进行检测和/或分离的标记或标签。
17.如权利要求1-16任一项的变体，其融合或缀合至半衰期延长模块。
18.如权利要求17的变体，其中该半衰期延长模块选自由下列组成的组：聚合物(例如聚乙二醇(PEG)，羟乙基淀粉(HES)，透明质酸，聚唾液酸)，非结构化(多)肽链(例如PAS，XTEN)，弹性蛋白样多肽(ELP)，血清蛋白(例如白蛋白)，血清蛋白结合分子(例如白蛋白结合域(ABD)，白蛋白结合脂肪酸)，抗体，免疫球蛋白，免疫球蛋白的Fc区/域和免疫球蛋白结合域。
19.一种融合分子，其包含如权利要求1-18任一项的人FGF21的变体及至少一种其他活性药物成分。
20.一种核酸分子，其编码如权利要求1-18任一项的人FGF21的变体或权利要求19的融合分子。
21.如权利要求20的核酸分子，其包含于载体中或整合至基因组中。
22.一种宿主细胞，其含有如权利要求20或21的核酸分子。
23.一种生产如权利要求1-18任一项的人FGF21的变体或如权利要求19的融合分子的方法，其包括培养如权利要求22的宿主细胞并从培养基分离该变体或融合分子。
24.一种药物组合物，其包含如权利要求1-18任一项的人FGF21的变体或如权利要求19的融合分子或如权利要求20或21的核酸分子或如权利要求22的宿主细胞，连同医药上可接受之载剂和/或赋形剂。
25.如权利要求24的药物组合物，其进一步包含至少一种其他活性药物成分。
26.如权利要求19的融合分子或如权利要求25的药物组合物，其中该至少一种其他活性药物成分选自由下列组成的组：胰岛素和胰岛素衍生物，GLP-1，GLP-1类似物和GLP-1受体激动剂，聚合物结合的GLP-1和GLP-1类似物，双重GLP-1/GIP激动剂，双重GLP-1/胰高血糖素受体激动剂，PYY3-36或其类似物，胰多肽或其类似物，胰高血糖素受体激动剂或拮抗剂，GIP受体激动剂或拮抗剂，胃饥饿素(ghrelin)拮抗剂或反向激动剂，Xenin及其类似物，DDP-IV抑制剂，SGLT-2抑制剂，双重SGLT-2/SGLT-1抑制剂，双胍类，噻唑烷二酮类，PPAR激动剂，PPAR调节剂，磺酰脲类，氯茴苯酸类，α-葡萄糖苷酶抑制剂，胰淀素及胰淀素类似物，GPR119激动剂，GPR40激动剂，GPR120激动剂，GPR142激动剂，TGR5激动剂，AMPK刺激物，AMPK激活剂，11-β-HSD抑制剂，葡萄糖激酶激活剂，DGAT抑制剂，蛋白酪氨酸磷酸酶1抑制剂，葡萄糖-6-磷酸酶抑制剂，果糖-1,6-二磷酸酶抑制剂，糖原磷酸化酶抑制剂，磷酸烯醇丙酮酸羧激酶抑制剂，糖原合酶激酶抑制剂，丙酮酸脱氢酶激酶抑制剂，CCR-2拮抗剂，葡萄糖转运蛋白-4调节剂，生长抑素受体3激动剂，HMG-CoA-还原酶抑制剂，贝特类，烟酸及其衍生物，烟酸受体1激动剂，ACAT抑制剂，胆固醇吸收抑制剂，胆汁酸结合物质，IBAT抑制剂，MTP抑制剂，PCSK9调节剂，LDL受体上调调节剂(肝选择性甲状腺激素受体β激动剂)，HDL升高化合物，脂质代谢调节剂，PLA2抑制剂，ApoA-I增强剂，胆固醇合成抑制剂，ω-3脂肪酸及其衍生物，用于治疗肥胖症的活性物质，CB1受体拮抗剂，MCH-1拮抗剂，MC4受体激动剂和部分激动剂，NPY5或NPY2拮抗剂，NPY4激动剂，β-3肾上腺素能受体激动剂，瘦蛋白或瘦蛋白模拟物，5HT2c受体激动剂，脂肪酶抑制剂，血管生成抑制剂，H3拮抗剂，AgRP抑制剂，三重单胺摄取抑制剂，MetAP2抑制剂，针对成纤维细胞生长因子受体4或阻抑素(prohibitin)靶向肽-1的产生的反义寡核苷酸，用于影响高血压、慢性心力衰竭或动脉粥样硬化的药物，血管紧张素II受体拮抗剂，双重血管紧张素受体阻断剂(ARB)，血管紧张素转化酶(ACE)抑制剂，血管紧张素转化酶2(ACE-2)活化剂，肾素抑制剂，肾素原抑制剂，内皮素转化酶(ECE)抑制剂，内皮素受体阻断剂，内皮素拮抗剂，利尿剂，醛固酮拮抗剂，醛固酮合酶抑制剂，α-阻断剂，α-2肾上腺素能受体拮抗剂，β-阻断剂，混合的α/β-阻断剂，钙拮抗剂/钙通道阻断剂(CBB)，双重盐皮质激素/CCB，中枢作用的抗高压药，中性内肽酶抑制剂，氨基肽酶-A抑制剂，血管肽(vasopeptide)抑制剂，双重血管肽抑制剂，脑啡肽酶(neprilysin)-ACE抑制剂，脑啡肽酶-ECE抑制剂，双重作用血管紧张素(AT)受体-脑啡肽酶抑制剂，双重AT1/内皮素-1(ETA)拮抗剂，晚期糖基化终末产物破坏剂，重组肾酶，血压疫苗，抗RAAS疫苗，AT1-或AT2-疫苗，具有抗高压响应的遗传多态性调节剂和血小板聚集抑制剂。
27.一种试剂盒，其包含如权利要求1-18任一项的人FGF21的变体或如权利要求19或26的融合分子或如权利要求20或21的核酸分子或如权利要求22的宿主细胞或如权利要求24-26任一项的药物组合物。
28.如权利要求1-18任一项的人FGF21的变体或如权利要求19或26的融合分子或如权利要求20或21的核酸分子或如权利要求22的宿主细胞或如权利要求24-26任一项的药物组合物，其用作药物。
29.如权利要求1-18任一项的人FGF21的变体或如权利要求19或26的融合分子或如权利要求20或21的核酸分子或如权利要求22的宿主细胞或如权利要求24-26任一项的药物组合物，其用于治疗选自由下列组成的组的疾病或病症：肥胖症，超重，代谢综合征，糖尿病，高血糖症，血脂异常，非酒精性脂肪性肝炎(NASH)和动脉粥样硬化。
30.如权利要求1-18任一项的人FGF21的变体或如权利要求19或26的融合分子或如权利要求20或21的核酸分子或如权利要求22的宿主细胞或如权利要求24-26任一项的药物组合物在制造用于治疗选自由下列组成的组的疾病或病症的药物中的用途：肥胖症，超重，代谢综合征，糖尿病，高血糖症，血脂异常，非酒精性脂肪性肝炎(NASH)和动脉粥样硬化。
31.一种用于治疗选自由下列组成的组的疾病或病症的方法：肥胖症，超重，代谢综合征，糖尿病，高血糖症，血脂异常，非酒精性脂肪性肝炎(NASH)和动脉粥样硬化，该方法包括将如权利要求1-18任一项的人FGF21的变体或如权利要求19或26的融合分子或如权利要求20或21的核酸分子或如权利要求22的宿主细胞或如权利要求24-26任一项的药物组合物施用于对其有需要的受试者。
32.用于如权利要求29的用途或用于如权利要求30的用途或如权利要求31的方法的变体或融合分子或核酸分子或宿主细胞或药物组合物，其中该疾病或病症是糖尿病。
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US20180371041A1 (en) 2018-12-27
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