CN117616038A - 突变型人nax蛋白和筛选方法 - Google Patents

突变型人nax蛋白和筛选方法 Download PDF

Info

Publication number
CN117616038A
CN117616038A CN202280038599.0A CN202280038599A CN117616038A CN 117616038 A CN117616038 A CN 117616038A CN 202280038599 A CN202280038599 A CN 202280038599A CN 117616038 A CN117616038 A CN 117616038A
Authority
CN
China
Prior art keywords
human
leu
ile
residues
diii
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202280038599.0A
Other languages
English (en)
Inventor
C·L·诺兰
J·M-D·帕安德
S·A·普莱斯
谢翰超
M·克尚萨克
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kobenhavns Universitet
Genentech Inc
Original Assignee
Kobenhavns Universitet
Genentech Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kobenhavns Universitet, Genentech Inc filed Critical Kobenhavns Universitet
Publication of CN117616038A publication Critical patent/CN117616038A/zh
Pending legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G01N2500/02—Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Cell Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Toxicology (AREA)
  • Food Science & Technology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Peptides Or Proteins (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

本公开涉及突变型人NaX离子通道(“NaX”)蛋白和使用突变型人NaX的筛选方法，所述筛选方法可用于鉴定调节人NaX的活性的分子。例如，在一些实施方案中，筛选方法涉及在潜在NaX调节剂的存在下对突变型人NaX进行离子通道测定。本公开还涉及充当人NaX的调节剂的分子以及用于检测此类分子的相关试剂盒。

Description

突变型人NAX蛋白和筛选方法
相关申请的交叉引用
本申请要求于2021年5月30日提交的美国临时申请号63/195,039的优先权，该美国临时申请全文以引用方式并入本文。
技术领域
本公开涉及突变型人NaX离子通道(“NaX”)蛋白和使用突变型人NaX的筛选方法，该方法可用于鉴定调节人NaX的活性的分子。例如，在一些实施例中，筛选方法涉及在潜在NaX调节剂的存在下，对突变型人NaX进行离子通道测定。本公开还涉及充当人NaX的调节剂的分子以及用于检测此类分子的相关试剂盒。
背景技术
原型电压门控钠(NaV)通道通过启动并传播动作电位在电信号传导中发挥关键作用。九种密切相关的哺乳动物通道亚型Nav1.1-NaV1.9在一系列组织中表达，在这些组织处，它们有助于包括神经系统功能、肌肉收缩和激素分泌在内的基本生理过程。第十种NaV通道样基因SCN7A在近三十年前被克隆，但非典型序列特征和无法记录来自这种NaV2.1蛋白的电压活化钠(Na+)电流引发了人们对其可能具有不同生理作用的猜测，因而将其命名为NaX。
与典型NaV通道相比，已提出NaX作为由细胞外Na+浓度活化的Na+通道而发挥作用。多种证据表明，NaX可能有助于Na+稳态1，2。NaX在专门监测血液组成的大脑区域中显示出受限的表达5。NaX敲除小鼠即便在脱水的情况下也摄入盐分，并且对血液Na+含量升高引起的高血压具有抵抗力5，6。在人类中，针对Nax的自体免疫导致慢性高钠血症，伴有受损的口渴感知和盐食欲(salt appetite)7。在外周，NaX调节上皮Na+通道(ENaC)上游的Na+稳态，并被建议作为治疗特应性皮炎和肥厚性瘢痕的靶标8。
Nax与NaV1.7通道具有最高的序列同一性(～55％)16，但它是NaV通道家族中趋异性最大的成员。在传统的NaV通道中，电压传感器结构域(VSD)包含4-8个沿S4区段保守的门控电荷残基，这些残基在膜去极化时经历向外移动以打开通道门(VSD1-VSD3)或启动快速失活(VSD4)。与此范例相反，据报道NaX对电压不敏感，并且包含数量减少的S4门控电荷残基。因此，NaX中电压传感器样结构域(VSLD)的功能和构象状态尚不清楚，并且VSLD如何促成通道门控尚不清楚。此外，相较于NaV，在NaX中趋异的DIII-DIV接头区段已知是NaV通道快速失活所需的。
发明内容
本公开涉及突变型人NaX离子通道(“NaX”)蛋白和使用突变型人NaX的筛选方法，这些方法可用于鉴定调节人NaX的活性的分子，并且涉及通过此类方法和用于进行这些方法的试剂盒鉴定的分子。例如，可以使用突变型人NaX蛋白和相关方法来不仅鉴定调节NaX活性的分子，而且测试已知结合至或调节NaV蛋白的分子，以测试该分子对于其靶标蛋白的选择性。在本文所述的使用突变型Nax的测定中鉴定的调节剂可用作人Nax的调节剂/候选调节剂。本发明人惊奇地发现突变型Nax蛋白，其允许通过模拟离子通道的开放、导电状态进行人Nax相关的离子通道测量。不受理论束缚，突变型Nax可以通过模拟例如离子通道蛋白的磷酸化和/或正常情况下允许Nax离子通道开放的其他相关生理学或结构变化来模拟Nax离子通道的开放、导电状态。本公开包括例如以下实施例中的任一项或其组合。
在一些实施例中，本文的公开内容涉及鉴定人NaX离子通道蛋白(NaX)调节剂的方法，该方法包括：(a)提供突变型人NaX，其中人NaX的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换；(b)在潜在Nax调节剂的存在下对突变型人Nax进行离子通道测定；(c)如果在潜在调节剂的存在下测定中的突变型人NaX的活性高于或低于在潜在调节剂不存在下的活性，则将该潜在调节剂鉴定为人NaX调节剂。在一些情况下，突变型人NaX包含来自人或哺乳动物Nav的DIII S1-S6区域的全部或部分，任选地其中经置换的人NaX的DIII结构域的至少一部分包含以下或由以下组成：(a)DIII(残基920-1200)；(b)DIII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)；(c)DIII VSD3、S4-S5接头和S5(残基920-1078)；(d)DIII和DII-DIII接头(残基733-1200)；(e)DIII和DIII-DIV接头(920-1237)；(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。在一些情况下，经人或哺乳动物NaV的DIII结构域的对应部分置换人NaX的DIII结构域的至少一部分不包含S5且/或不包含S6。在一些情况下，经人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的至少一部分不包含S1、S2、S3、S4和/或S4-S5接头中的任一者。在一些情况下，人Nax的DIII结构域的至少一部分经哺乳动物Nav1.1、哺乳动物Nav1.2、哺乳动物Nav1.3、哺乳动物Nav1.4、哺乳动物Nav1.5、哺乳动物Nav1.6、哺乳动物Nav1.7、哺乳动物Nav1.8、哺乳动物Nav1.9、人Nav1.1、人Nav1.2、人Nav1.3、人Nav1.4、人Nav1.5、人Nav1.6、人Nav1.7、人Nav1.8或人Nav1.9的对应部分置换。例如，在一些情况下，人NaX的DIII结构域的至少一部分经人Nav1.7的对应部分置换。在一些情况下，突变型人NaX是嵌合体构建体2、嵌合体构建体3、嵌合体构建体7、嵌合体构建体9、嵌合体构建体11、嵌合体构建体15或嵌合体构建体19，或包含SEQ ID No：3、4、8、10、12、16或20中的任一者的氨基酸序列。
本公开还涵盖鉴定人NaX离子通道蛋白(NaX)调节剂的方法，该方法包括：(a)提供突变型人NaX，其包含以甘氨酸、脯氨酸或极性或带电残基对人NaX中的两个、三个或四个S6α螺旋中的每个S6α螺旋上的至少一个残基的取代；(b)在潜在调节剂的存在下对突变型人NaX进行离子通道测定；(c)如果在潜在调节剂的存在下测定中的突变型人NaX的活性高于或低于在潜在调节剂不存在下的活性，则将该潜在调节剂鉴定为人NaX调节剂。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的两个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的三个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的所有四个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的至少三个S6α螺旋上的氨基酸残基的一个取代。在一些情况下，突变型人NaX包含在SEQ ID NO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含在SEQ ID NO：1的以下区段中的至少三个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX在SEQ ID NO：1的以下区段中的每个区段内包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对在残基L390、F724、I1189和I1492中的两者或更多者处的氨基酸残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对在残基L390、F724、I1189和I1492中的三者或更多者处的氨基酸残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基L390、F724和I1189的取代或对氨基酸残基F724、I1189和I1492的取代。在一些情况下，相较于野生型人Nax，突变型人Nax包含取代F724Q、I1189T和I1492T。在一些情况下，相较于野生型人NaX，该人NaX包含取代L390E、I1189E和I1492E。
在任何上述方法中，离子通道测定也可以在突变型人NaX的经鉴定的调节剂(例如，本文经鉴定的调节剂或在本文方法的进行中所鉴定的其他调节剂)的存在下进行。在一些此类情况下，突变型人NaX的经鉴定的调节剂是河豚毒素(TTX)、奎尼丁(quinidine)、氟卡尼(flecainide)、丁卡因(tetracaine)或利多卡因(lidocaine)中的一者或多者。
在任何上述方法中，潜在调节剂可以是肽或大环或抗体。例如，在一些情况下，潜在调节剂是小分子。在一些情况下，小分子可以是例如河豚毒素(TTX)、奎尼丁、氟卡尼、丁卡因或利多卡因的衍生物。
任何上述方法也可进一步包括确定在(c)部分中鉴定的潜在调节剂对突变型人NaX和野生型人NaX中的任一者或两者的结合亲和力。在一些情况下，潜在调节剂以10μM或更少、10μM至50nM、10μM至500nM、1μM或更少、1μM至50nM、或100nM或更少的EC50或IC50，与突变型人NaX和野生型人NaX中的任一者或两者结合。在一些情况下，EC50或IC50可以在ELISA测定中确定。
在任何上述方法中，离子通道测定可以是膜片钳或自动化膜片钳测定、离子通量测定、或离子敏感或电压敏感染料测定。在任何上述方法中，在(c)部分中鉴定的潜在调节剂可以使该测定中的突变型人NaX的活性降低至少25％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％或至少90％。在一些情况下，在(c)部分中鉴定的潜在调节剂以10nM至500μM，50nM至500μM，10nM至50μM，100nM至500μM，100nM至50μM，1-500μM，1-50μM，10-500μM或50-250μM的半最大浓度使该测定中的突变型人NaX的活性降低。在其他情况下，在(c)部分中鉴定的潜在调节剂使该测定中的突变型人NaX的活性增加。
本公开还涉及通过本文所述的方法鉴定的人NaX调节剂，其可以任选地为肽、大环、小分子或抗体。
本公开还涵盖突变型人Nax离子通道(Nax)蛋白，其中人Nax的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换。在一些情况下，突变型人Nax包含来自人或哺乳动物Nav的DIIIS1-S6区域的全部或部分，任选地其中经置换的人NaX的DIII结构域的至少一部分包含以下或由以下组成：(a)DIII(残基920-1200)；(b)DIII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)；(c)DIII VSD3、S4-S5接头和S5(残基920-1078)；(d)DIII和DII-DIII接头(残基733-1200)；(e)DIII和DIII-DIV接头(920-1237)；(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。在一些情况下，经人或哺乳动物NaV的DIII结构域的对应部分置换人NaX的DIII结构域的至少一部分不包含S5且/或不包含S6。在一些情况下，经人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的至少一部分不包含S1、S2、S3、S4和/或S4-S5接头中的任一者。在一些情况下，人NaX的DIII结构域的至少一部分经哺乳动物Nav1.1、哺乳动物Nav1.2、哺乳动物Nav1.3、哺乳动物Nav1.4、哺乳动物Nav1.5、哺乳动物Nav1.6、哺乳动物Nav1.7、哺乳动物Nav1.8、哺乳动物Nav1.9、人Nav1.1、人Nav1.2、人Nav1.3、人Nav1.4、人Nav1.5、人Nav1.6、人Nav1.7、人Nav1.8或人Nav1.9的对应部分置换。例如，在一些情况下，人NaX的DIII结构域的至少一部分经人Nav1.7的对应部分置换。在一些情况下，突变型人NaX是嵌合体构建体2、嵌合体构建体3、嵌合体构建体7、嵌合体构建体9、嵌合体构建体11、嵌合体构建体15或嵌合体构建体19，或包含SEQ ID No：3、4、8、10、12、16或20中的任一者的氨基酸序列。
本公开进一步包括突变型人NaX离子通道(NaX)蛋白，其包含以甘氨酸、脯氨酸或极性或带电残基对两个、三个或四个S6α螺旋中的每个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的两个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的三个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的所有四个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的至少三个S6α螺旋上的氨基酸残基的一个取代。在一些情况下，突变型人Nax包含在SEQ ID NO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含在SEQ ID NO：1的以下区段中的至少三个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX在SEQ ID NO：1的以下区段中的每个区段内包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对在残基L390、F724、I1189和I1492中的两者或更多者处的氨基酸残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对在残基L390、F724、I1189和I1492中的三者或更多者处的氨基酸残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基L390、F724和I1189的取代或对氨基酸残基F724、I1189和I1492的取代。在一些情况下，相较于野生型人NaX，突变型人NaX包含取代F724Q、I1189T和I1492T。在一些情况下，相较于野生型人NaX，该人NaX包含取代L390E、I1189E和I1492E。
本公开进一步包括确定调节人离子通道蛋白活性的测试分子是否调节人Nax离子通道(NaX)蛋白的活性的方法，该方法包括：(a)提供突变型人NaX，其中人NaX的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(Nav)的DIII结构域的对应部分置换；(b)在测试分子的存在下对该突变型人NaX进行离子通道测定；以及(c)如果在测试分子的存在下该测定中的突变型人Nax的活性高于或低于在测试分子不存在下的活性，则将该测试分子确定为突变型人NaX调节剂；以及任选地，(d)如果根据(c)部分，测试分子不是人NaX调节剂，则选择该测试分子进行额外筛选。
本公开额外地包括确定调节人离子通道蛋白活性的测试分子是否调节人NaX离子通道(NaX)蛋白的活性的方法，该方法包括：(a)确定测试分子调节人离子通道蛋白的活性；(b)提供突变型人NaX，其中人NaX的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换；(c)在测试分子的存在下对该突变型人NaX进行离子通道测定；以及(d)如果在测试分子的存在下该测定中的突变型人NaX的活性高于或低于在测试分子的不存在下的活性，则将该测试分子确定为突变型人NaX调节剂；以及任选地，(e)如果根据(d)部分，测试分子不是人NaX调节剂，则选择该测试分子进行额外筛选。
在上述两种额外方法中，在一些情况下，突变型人NaX包含来自人或哺乳动物NaV的DIII S1-S6区域中的全部或部分，任选地，其中经置换的人NaX的DIII结构域的至少一部分包含以下或由以下组成：(a)DIII(残基920-1200)；(b)DiII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)；(c)DiII VSD3、S4-S5接头和S5(残基920-1078)；(d)DIII和DII-DIII接头(残基733-1200)；(e)DIII和DIII-DIV接头(920-1237)；(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。在一些情况下，经人或哺乳动物NaV的DIII结构域的对应部分置换人NaX的DIII结构域的至少一部分不包含S5且/或不包含S6。在一些情况下，经人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的至少一部分不包含S1、S2、S3、S4和/或S4-S5接头中的任一者。在一些情况下，人NaX的DIII结构域的至少一部分经哺乳动物Nav1.1、哺乳动物Nav1.2、哺乳动物Nav1.3、哺乳动物Nav1.4、哺乳动物Nav1.5、哺乳动物Nav1.6、哺乳动物Nav1.7、哺乳动物Nav1.8、哺乳动物Nav1.9、人Nav1.1、人Nav1.2、人Nav1.3、人Nav1.4、人Nav1.5、人Nav1.6、人Navl.7、人Nav1.8或人Nav1.9的对应部分置换。在一些此类情况下，人Nax的DIII结构域的至少一部分经人Nav1.7的对应部分置换。在一些情况下，突变型人NaX是嵌合体构建体2、嵌合体构建体3、嵌合体构建体7、嵌合体构建体9、嵌合体构建体11、嵌合体构建体15或嵌合体构建体19，或包含SEQ ID No：3、4、8、10、12、16或20中的任一者的氨基酸序列。
本公开还包括确定调节人离子通道蛋白活性的测试分子是否调节人NaX离子通道(NaX)蛋白的活性的方法，该方法包括：(a)提供突变型人NaX，其包含以甘氨酸、脯氨酸或极性或带电残基对人NaX的两个、三个或四个S6α螺旋中的每个S6α螺旋上的至少一个残基的取代；(b)在测试分子的存在下对突变型人Nax进行离子通道测定；(c)如果在测试分子的存在下该测定中的突变型人NaX的活性高于或低于在潜在测试分子不存在下的活性，则将该测试分子确定为人NaX调节剂；以及任选地，(d)如果根据(c)部分，测试分子不是人NaX调节剂，则选择该测试分子进行额外筛选。
本公开进一步涵盖确定调节人离子通道蛋白活性的测试分子是否也调节人NaX离子通道(NaX)蛋白的活性的方法，该方法包括：(a)确定测试分子调节人离子通道蛋白的活性；(b)提供突变型人NaX，其包含以甘氨酸、脯氨酸或极性或带电残基对人NaX的两个、三个或四个S6α螺旋中的每个S6α螺旋上的至少一个残基的取代；(c)在测试分子的存在下对突变型人Nax进行离子通道测定；(d)如果在测试分子的存在下该测定中的突变型人NaX的活性高于或低于在潜在测试分子不存在下的活性，则将该测试分子确定为人NaX调节剂；以及任选地，(e)如果根据(d)部分，测试分子不是人NaX调节剂，则选择该测试分子进行额外筛选。
在上述两种方法中的任一项中，在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6αα螺旋中的两个S6αα螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6αα螺旋中的三个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的所有四个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的至少三个S6α螺旋上的氨基酸残基的一个取代。在一些情况下，突变型人NaX包含在SEQ ID NO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含在SEQ ID NO：1的以下区段中的至少三个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX在SEQ ID NO：1的以下区段中的每个区段内包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对在残基L390、F724、I1189和I1492中的两者或更多者处的氨基酸残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对在残基L390、F724、I1189和I1492中的三者或更多者处的氨基酸残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基L390、F724和I1189的取代或对氨基酸残基F724、I1189和I1492的取代。在一些情况下，相较于野生型人NaX，突变型人NaX包含取代F724Q、I1189T和I1492T。在一些情况下，相较于野生型人NaX，该人NaX包含取代L390E、I1189E和I1492E。
本公开还包括通过上述方法鉴定的分子，其任选地可以是肽、大环、小分子或抗体。
本公开还包括分子复合物，其包含如本文所述的嵌合型或突变型人NaX蛋白以及该蛋白的调节剂或该蛋白的潜在调节剂。
本公开还包括用于测定人NaX的活性或用于鉴定人NaX调节剂和/或与人NaX结合的分子的试剂盒，其包括如本文所述的突变型人NaX，并且进一步包括以下中的至少一者：一种或多种进行离子通道测定的试剂、该突变型人NaX的调节剂，以及使用说明。
附图说明
图1A-1H显示人NaX的表征和β3-NaX通道复合物的整体结构。图1A显示来自表达人NaX或NaV1.7的非洲爪蟾(Xenopus laevis)卵母细胞的代表性电流。图1B显示来自表达NaX的卵母细胞响应于所指示化合物的细胞外施加的代表性电流。图1C显示来自表达NaX以及与NaV或CaV通道辅助亚基、Na+/K+-ATP酶亚基和突触相关蛋白97(SAP97)共表达的卵母细胞在所指示的细胞外Na+浓度的存在下的代表性电流。图1A-1C的电压程序如下：NaX：在+80到-100mV之间阶跃，从0 mV的HP开始以20mV为增量；NaV1.7：在-80与+65mV之间去极化阶跃，从-100 mV的HP开始，以5mV为增量。图1D显示图1A-1C中进行的独立实验的数据汇总。图1E显示来自表达人NaX或NaV1.7的小鼠Neuro-2细胞响应于细胞外Na+浓度(HP＝-60mV)变化的代表性电流，如所指示，或电压：在-60至+60mV之间去极化阶跃，从-100mV的HP开始，以10mV为增量。图1F显示如图1E中进行的独立实验的数据汇总。图1G显示从Neuro-2细胞中提取的蛋白质的总裂解物和表面部分的蛋白质印记，用于以探针探测所指示的蛋白质。图1H显示β3-NaX通道复合物的侧视图和细胞外视图。指示了近似的膜边界。DI、DII、DIII和DIV分别以绿色、蓝色、橙色和粉红色着色，且β3亚基以灰色表面表示。
图2A-2E显示，NaX孔模块的结构表明非导电状态。图2A显示作为灰色表面显示的NaX孔体积，其中DII和DIV在卡通渲染中显示(为清楚起见，省略了DI和DIII)。图2B显示S6螺旋的视图，其中侧链以棒状和半透明表面表示形式衬砌活化门。正交视图提供更广阔的视角，DIII和DIV以橙色和粉红色着色，来自DIII-DIV接头的IFI基序(绿色，虚线框)以棒状和半透明表面表示形式显示。图2C显示透过孔模块切片的正交视图，突出显示横向开窗和所结合的脂质。穿过S6门的磷脂酰乙醇胺以紫色棒状表示。在侧视图中，为清楚起见，已移除前景中的脂质。图2D中的视图类似于中间图C，但在以蓝色网格表示的模拟脂质周围显示cryo-EM图。图2E显示来自非洲爪蟾卵母细胞的代表性电流，该卵母细胞表达人NaX、人NaV1.7、或双结构域和单结构域调换的NaV1.7-NaX嵌合体，包括本文表4中所示的嵌合体构建体1-7。在+80到-100mV之间阶跃，从0 mV的HP开始，以20mV为增量。
图3A-3F显示携带靶向孔润湿S6突变的人NaX的表征。图3A显示衬砌S6门的所靶向的疏水侧链的位置和放大视图。图3B显示来自表达NaX-QTT构建体的非洲爪蟾卵母细胞在所指示的电压程序下的代表性电流。图3C显示来自表达所指示的NaX构建体的卵母细胞的代表性电流。图3D显示来自表达NaX-EEE构建体的卵母细胞的代表性电流。图3C-3D的电压程序如上图2E中所示。图3E显示来自表达NaX-QTT构建体且共表达所指示的NaV辅助亚基的卵母细胞的代表性电流。电压程序如上。图3F显示图2B-E中进行的独立实验的数据汇总。
图4A-4C显示NaX选择性过滤器的结构和表征。图4A显示作为棒状显示的NaX DENA-基序残基侧链。图4B上图显示在NaV通道中的选择性过滤器周围形成相互作用网络的NaX侧链和类似残基，以棒状表示(NaV1.7，PDB 6J8J)显示。图4B下图显示NaX和NaV1.7选择性过滤器的相同视图(如图4B上图中)，但作为静电表面渲染。为清楚起见，中央腔和活化门被排除。图4C显示DI-DIV界面的叠加和放大视图，比较NaX与NaV1.7(灰色，PDB 6J8J)结构，其中所选择的侧链显示为棒状。图4D显示来自表达带有C末端GFP-Flag标签的人NaX-QTT的HEK293细胞在生理(图4D，左图)或仅NMDG+细胞外溶液(图4D，中间图)中的代表性电流。在+80到-100mV之间阶跃，从0 mV的HP开始，以20mV为增量。图4D右图显示来自独立实验的I-V曲线数据汇总。图4E左图显示来自表达人NaX-QTT的HEK293细胞在具有所指示的单价阳离子的细胞外溶液中的代表性电流。施加从-80到+80mV的电压斜坡。图4E右图显示从独立实验中测量的反转电位和渗透性比率的汇总。
图5A-5C显示人NaX-QTT通道的药理学。图5A左图和中图显示来自表达人NaX-QTT的HEK293细胞在具有所指示的细胞外单价阳离子且具有或不具有所指示量的CaCl2的细胞外溶液中的代表性I-V电流。施加从-80到+80mV的电压斜坡。图5A右图显示在80mV时通过所指示的Ca2+浓度达到的外向Na+阻断的百分比。图5B显示当从0到+80mV阶跃时，表达人NaX-QTT的非洲爪蟾卵母细胞在具有或不具有所指示的二价和三价阳离子(单位是mM)的标准细胞外溶液中的代表性电流。图5C显示当从0到+80mV(左图)或从0到-100mV(右图)阶跃时，表达人NaX-QTT的卵母细胞在添加或不添加所指示的阻断剂的标准细胞外溶液中的代表性电流。图5C中图显示从独立实验中测量的电流汇总。
图6A-6E显示非典型NaX电压传感器样结构域的结构。图6A显示带有门控电荷(蓝色)、HCS(绿色)和ENC/INC(红色)侧链的VSLD1，以棒状表示。插图突出显示NaX中的一个独特脯氨酸，并与NaV1.7VSD1(PDB 6J8J)叠加以进行比较。图6B显示如图6A中渲染的VSLD2。插图突出显示His579-S4相互作用(图6B，上图)和S4-S5接头偏移(图6B，下图)，与NaV1.7DII(PDB 6J8J)叠加以进行比较。图6C显示如图6A中渲染的VSLD3。插图突出显示来自S3螺旋中的Phe1011。图6D显示了VSLD4，如图6A中渲染。插图显示NaX与NaV1.7VSD4(PDB6J8J)之间的S4/S4-S5接头区的比较，其中门控电荷残基的Cα表示为蓝色球体。图6E显示表达NaV1.7和各种NaV1.7-NaX-VSLD的非洲爪蟾卵母细胞响应于在-80与+65mV之间去极化阶跃的代表性电流，该阶跃从-100mV的HP开始，以5mV为增量。
图7A-7F显示人NaX在不同表达系统中的功能评估。图7A显示从表达所指示的构建体的HEK293T细胞中提取的蛋白质的总裂解物和表面部分的蛋白质印记。图7B显示来自表达人NaX或NaV1.7(带有C末端GFP和Flag标签)的HEK293T细胞在所指示的电压程序下的代表性电流。图7C显示从表达所指示的构建体的非洲爪蟾卵母细胞中提取的蛋白质的总裂解物和表面部分的蛋白质印记。图7D显示来自表达人NaX或NaV1.7的卵母细胞在所指示的电压程序下的代表性电流。图7E显示从表达所指示的构建体的小鼠Neuro-2A细胞中提取的蛋白质的总裂解物和表面部分的蛋白质印记。图7F显示表达人NaX(*，带有C末端GFP和Flag标签)的Neuro-2A细胞在所指示的细胞外Na+浓度(HP＝-60mV)下的代表性电流。提供了单个细胞的密封电阻(Rm)。
图8A-8D显示人NaX在非洲爪蟾卵母细胞中的功能评估。图8A左图和8B左图显示来自表达人NaX的非洲爪蟾卵母细胞在所指示的药理学调节剂的存在下在细胞外溶液中的代表性电流。图8A右图显示I-V图，在+80和-100mV之间去极化阶跃，从0mV的HP开始，以20mV为增量。图8B左图显示I-V图，在-100和+100mV之间去极化阶跃，从-100mV的HP开始，以20mV为增量。图8C左图和8D左图显示来自表达带有共表达的所指示的蛋白质的人NaX的卵母细胞的代表性电流。图8C右图显示I-V图，在+80和-100mV之间阶跃，从0mV的HP开始，以20mV为增量。图8D显示I-V图，在-100和+100mV之间去极化阶跃，从-100mV的HP开始，以20mV为增量。
图9显示NaX与NaV通道的多序列比对。显示了NaX结构域边界以供参考。NaX门控电荷用星号表示，且选择性过滤器残基用方框表示。NaX DIII-DIV接头IFI基序由三个连续的星号表示。
图10A-10C显示β3-NaX通道样品纯化。图10A显示β3-NaX表达、纯化和重组方案。图10B显示脂质纳米盘(MSP1E3D1)中β3-NaX样品的尺寸排阻色谱洗脱曲线。图10C显示尺寸排阻色谱洗脱级分的SDS-PAGE分析。
图11A-11L显示cryo-EM数据处理工作流程。图11A显示β3-NaX-纳米盘样品的代表性cryo-EM显微照片。图11B显示具有代表性的2D类平均值。图11C显示热图，其显示最终重建中指定粒子取向的总体分布。图11D显示基于两个半数据集之间的傅里叶壳层相关性(FSC)的全局分辨率估计。图11E显示通过局部分辨率着色的最终3D重建的等值面渲染，如通过窗口化FSC估计。图11F显示针对DI的跨膜区显示的cryo-EM图。图11G显示β3-NaX-洗涤剂(GDN)样品的代表性cryo-EM显微照片。图11H显示具有代表性的2D类平均值。图11I显示热图，其显示最终重建中指定粒子取向的总体分布。图11J显示基于两个半数据集之间的傅里叶壳层相关性(FSC)的全局分辨率估计。图11K显示通过局部分辨率着色的最终3D重建的等值面渲染，如通过窗口化FSC估计。图11L显示针对DI的跨膜区显示的cryo-EM图。
图12A-B显示cryo-EM数据处理工作流程。图12A显示β3-NaX-纳米盘样品的cryo-EM数据处理工作流程的示意图。图12B显示β3-NaX-洗涤剂样品的cryo-EM数据处理工作流程的示意图。
图13A-13F显示整体NaX结构、β3相互作用以及与NaV通道的比较。图13A显示β3-NaX-纳米盘复合物的cryo-EM重建。NaX NTD(N-末端结构域)位于细胞内侧，且β3-亚基ECD(细胞外结构域)位于细胞外侧。指示了近似的膜边界。图13B显示3-NaX复合物的侧视图。插图突出显示3与NaX的相互作用。图13C-13E显示3-NaX与人NaV1.2(PDB 6J8E)、NaV1.4(PDB6AGF)、NaV1.5(PDB 6UZ0)和NaV1.7(PDB 6J8J)结构以各种透视角度进行比对的特写视图。图13F显示中央腔的特写视图，突出显示与Nav1.7(PDB 6J8J)叠加的NaX的DIV W1484侧链。NaV1.7的DIV Phe以棒状表示。
图14A-14F显示NaX的DIII-DI接头、DIII嵌合通道和脂质结合孔结构。图14A显示来自β3-NaX-纳米盘结构的DIII和DIV的侧视图和细胞内视图，其中DIII-DIV接头的IFI-基序以球体显示，且显示NaV1.7(PDB 6J8J)的等效区用于比较。图14B显示DIII-DIV接头区与IFI/IFM-基序的多序列比对。图14C显示来自表达野生型或IFI>QQQ突变型人NaX的卵母细胞的代表性电流，其中在+80与-100mV之间电压阶跃，从0mV的HP开始，以20mV为增量，或在-100与+120mV之间去极化阶跃，从-100mV的HP开始，以20mV为增量。I-V图如图14C右图中所示。图14D显示来自表达各种DIII人NaV1.7-NaX通道嵌合体的卵母细胞响应于+80与-100mV之间的电压阶跃的示意图和代表性电流，该电压阶跃从0 mV的HP开始，以20mV为增量。图14D右图，各种DIII NaV1.7-NaX嵌合体在+80mV(上图)和-100mV(下图)时的最大电流振幅。图14E显示β3-NaX-纳米盘S6-门结构的侧视图，其中衬砌门的侧链以棒状和半透明表面表示形式显示，并且指定的磷脂酰乙醇胺以棒状显示。图14F显示孔的顶视图，比较基于β3-NaX-纳米盘和β3-NaX-洗涤剂的结构，其中该孔中的指定脂质以棒状表示形式显示。cryo-EM图围绕β3-NaX-GDN结构的网格表示中的模拟脂质显示。
图15A-15C显示NaX与NaV1.7选择性过滤器的比较。图15A显示NaX和NaV1.7(PDB6J8J)选择性过滤器的等效视图。NaV1.7中结合的河豚毒素显示具有选择相互作用。图15B显示来自表达野生型或突变型人NaV1.7和NaX通道构建体的非洲爪蟾卵母细胞在115mM细胞外Na+、K+、Li+或Cs+的存在下的代表性电流。显示在-50与+50mV之间的阶跃，从-100mV的HP开始，以10mV阶跃。图15C显示来自表达人NaX-QTT(含有C末端GFP-Flag标签)的HEK293细胞在氯(Cl-)离子(经甲磺酸盐MS-取代)存在(左)或不存在(右)下响应于从-80到+80mV的电压斜坡的代表性电流。
图16A-16E显示NaX电压传感器样结构域的结构和比较。图16A显示NaV1.7VSD4(PDB6J8J)和NaX VSLD4的等效视图，其中门控电荷侧链以棒状显示，且R3-R5经标记用于参考。图16B显示野生型NaV1.7和各种突变通道的活化和稳态失活的电压依赖性。活化，在-80和+65mV之间的去极化阶跃之后获得电流，该去极化阶跃从-100mV的HP开始，以5mV为增量。失活，在从-100mV的HP开始的一系列调节脉冲(在-120与-10mV之间，以5mV为增量，500ms)之后，在0或-20mV(25ms)的测试脉冲期间获得电流。归一化的活化和失活曲线显示在电流轨迹的右侧。图16C显示NaX VSDL3与KCNQ1 VSD(PDB 6UZZ)的叠加，其中NaX门控电荷和HCS显示为棒状。NaX S3 F1101和来自KCNQ1的对应S3丝氨酸以表面表示形式显示；请注意，该位置在所有人NaV通道中也被保守为丝氨酸。图16D显示NaXVSDL4的视图，突出显示与人NaV通道VSD4序列相比时独特的脯氨酸侧链。图16E显示野生型NaV1.5和三脯氨酸VSD4突变通道的活化和稳态失活的电压依赖性。活化，在-80和+40mV之间的去极化阶跃之后获得电流，该去极化阶跃从-100mV的HP开始，以5mV为增量。失活，在从-100mV的HP开始的一系列调节脉冲(在-120与-10mV之间，以5mV为增量，500ms)之后，在0或-20mV(25ms)的测试脉冲期间获得电流。归一化的活化和失活曲线显示在电流轨迹的右侧。
图17A-17B显示NaX选择性过滤器和S6门区域的结构和比较。图17A显示选择性过滤器的特写视图，其中在NaX中具有推定的离子结合位点(DEE基序)，显示了cryo-EM图(网格)的一部分和模拟为球体的Na+离子。显示在NaV1.2(PDB 6J8E)和NaVPaS(PDB 6NT4)中的DEE基序结合位点中指定的Na+离子用于比较。图17B显示NaX S6)门的细胞内视图，显示了Tyr1491(S6)和所结合的磷脂酰乙醇胺并且与NaV1.4(假设处于非导电、失活状态；PDB6AGF和药物结合的NaV1.5(假设处于非导电、药物阻断状态；PDB 6UZ0)比较。指定的脂质、洗涤剂和药物分子(PE、GDN或氟卡尼)分别显示为棒状表示。
图18显示表达构建体1-25的非洲爪蟾卵母细胞的电流振幅。(*p＜0.05；**p＜0.01；****p＜0.0001；针对NaX的单因素方差分析(one-way ANOVA)，Dunnett检验。)
图19A-B显示NaX通道的结构。在图19A中，显示了DIII+DIII-IV接头(920-1237)。在图19B中，显示了S6门区域。
图20显示NaX通道S6区段(部分)与NaV通道的多序列比对。
具体实施方式
I.定义
除非另有定义，否则与本发明结合使用的科学和技术术语应具有本领域普通技术人员通常理解的含义。如根据本公开所使用的，除非另外指出，否则以下术语应理解为具有以下含义：
在本申请中，除非另外陈述，否则所用的“或”意指“和/或”。在多项从属权利要求的上下文中，使用“或”仅以替代形式重新提及多于一个前述独立或从属权利要求。同样，除非另外具体说明，否则诸如“要素”或“组分”等术语涵盖包含一个单元的要素和组分以及包含多于一个亚单元的要素和组分两种情况。
如本文中所使用的过渡术语“基本上由…组成”，当涉及所要求保护的方法的步骤时，表示该方法除了规定的那些将实质上影响该方法的基本和新颖特征的步骤之外，不包括其他步骤。如本文中所使用的过渡术语“基本上由…组成”，当涉及诸如试剂盒的组合物或产品时，表示其不包含除了规定的那些将实质上影响其基本和新颖特征的组分以外的其他组分。
如本文所用，除非上下文另外明确指出，否则单数形式的“一种(a)”、“一个(an)”和“该(the)”包括复数指示内容。
如本文所述，除非另外指示，否则任何浓度范围、百分比范围、比率范围或整数范围应理解为包括在所引述范围及(在适当时)其分数部分(诸如整数的十分之一和百分之一)内的任何整数值。
单位、前缀、和符号以其国际单位制(Système International de Unite，SI)接受的形式来表示。数字范围包括定义范围的数字。本文提供的标题不是对本公开的各个方面的限制，可以通过引用整个说明书来获得本公开的各个方面。据此，通过整体引用说明书更完整地定义了下面直接定义的术语。
除非另外明确描述，否则本文所述的所有蛋白均为人蛋白，如在“哺乳动物NaV”或类似短语中。“哺乳动物”蛋白包括人蛋白以及来自其他哺乳动物物种的等效蛋白。
如本文所述，“Nav”蛋白或“Nav”或“NaV”蛋白是电压门控钠通道蛋白家族的成员。人和其他哺乳动物中的NaV家族成员的示例包括Nav1.1、Nav1.2、Nav1.3、Nav1.4、Nav1.5、Nav1.6、Nav1.7、Nav1.8和Nav1.9。人NaV家族成员蛋白包含四个结构域(DI、DII、DIII和DIV)，每个结构域包含六个跨膜α螺旋(S1、S2、S3、S4、S5和S6)，在一些情况下，在一个结构域内的某些α螺旋之间以及在四个结构域之间具有环或接头，共有24个跨膜α螺旋。
如本文所述，“NaX”离子通道蛋白或“Nax”或“NaX”蛋白是电压门控钠通道蛋白家族的成员，也称为Nav2.1或scn7a。除非另有说明，否则“NaX”蛋白指代“人NaX”蛋白。据报道，人NaX蛋白在中枢神经系统并且还在皮肤和其他上皮细胞中作为钠传感器。野生型人NaX的氨基酸序列在UniProt数据库中提供(登录号Q01118)，并在本文中作为SEQ ID NO：1提供。人NaX蛋白还包括该蛋白的天然存在的变体，其示例也可在例如UniProt数据库中的编号Q01118下找到。与人NaV蛋白一样，人NaX包含四个结构域(DI、DII、DIII和DIV)，每个结构域包含六个跨膜α螺旋(S1、S2、S3、S4、S5和S6)。
如本文所用，“突变型人NaX”指代，相较于SEQ ID NO：1的野生型NaX蛋白，包含至少一个经工程化的氨基酸取代、插入或缺失的人NaX蛋白。在一些情况下，“突变型人NaX”包含至少一个氨基酸取代，其中野生型人NaX的至少一个残基经哺乳动物或人NaV的等效/对应残基取代。在一些情况下，野生型人NaX的区域或完整结构域经哺乳动物或人NaV的等效区域或结构域取代，例如，DIII和/或DIII-DIV接头中的一些或全部。在此类情况下，突变型人NaX在本文中可替代地指代为“嵌合型人NaX”或“嵌合NaX”或“嵌合体”，因为它含有来自不同蛋白质、人或哺乳动物NaV的氨基酸序列片段。
本文中的术语“嵌合”，当指代蛋白质时，意指该蛋白质由来自多于一种天然蛋白质的氨基酸序列构成。例如，如上所述，如本文所用，“嵌合型人NaX”指代一种类型的突变型人NaX，其中NaX蛋白的至少一个区域已经与来自人NaV家族成员蛋白的对应区域交换。
本文中的术语“嵌合”，当指代蛋白质时，意指该蛋白质由来自多于一种天然蛋白质的氨基酸序列构成。例如，本文所用的“嵌合人NaX”指代一种类型的突变型人NaX，其中NaX蛋白的至少一个区域已经与来自NaV家族成员蛋白(诸如来自哺乳动物或人NaV家族成员蛋白，诸如人或哺乳动物Nav1.1、Nav1.2、Nav1.3、Nav1.4、Nav1.5、Nav1.6、Nav1.7、Nav1.8或Nav1.9)的对应区域交换。在一些情况下，交换区域可以是完整结构域，或结构域内的一个或多个单独的跨膜α螺旋或接头区或环，或结构域内的α螺旋和/或环的一部分。
如本文所用，术语“对应”或“等效物”指代来自人或哺乳动物NaV蛋白的残基或区域时可互换使用，其在制备突变型或嵌合型人NaX时置换从人NaX中缺失的残基或区域。如本文所用，“对应”或“等效”残基或区域是当正确折叠时位于两个蛋白质内相同位置的那些残基或区域。在一些情况下，可以使用针对两个蛋白质的序列比对和结构信息来鉴定此类对应或等效区域或氨基酸残基。
如本文所用，“调节剂”指代能够改变蛋白质行为的分子，诸如离子通道蛋白，如人NaX。例如，在一些情况下，调节剂可以通过与蛋白质结合来改变靶标蛋白质的行为。本文的调节剂可用于增加或降低蛋白质活性，例如，蛋白质调节离子流过细胞膜的程度。例如，降低人NaX活性的调节剂是人NaX活性的“抑制剂”。例如，增加人NaX活性的调节剂是人Nax活性的“活化剂”。在一些实施例中，调节剂可以仅在某些条件下(诸如在某些离子的存在下或在某些二级分子的存在下)调节人NaX的活性。
如本文所用，人NaX的“潜在调节剂”是待测试以确定其是否作为人Nax的调节剂的分子。
本文中，“离子通道测定”指代用于测量离子通道蛋白(诸如电压门控钠通道)的活性的测定。本领域经常使用多种测定和测定形式，并且许多以自动化形式提供用于高通量筛选(HTS)分析。示例包括离子通量测定，例如，使用放射性离子诸如放射性Na+离子，离子敏感或电压敏感染料测定诸如荧光测定，例如使用其荧光信号随离子浓度变化而增加或减少的荧光指示剂分子，以及各种类型的膜片钳测定。此类测定可以直接或间接地测量在各种条件下穿过包含离子通道蛋白的膜的离子电流的变化。在一些情况下，此类测定用于评估离子通道蛋白在潜在或经鉴定的调节剂存在或不存在下的“活性”。术语“活性”在这种意义上意指最广泛的意义，因为这些不同的测定直接或间接地并通过测量不同的参数诸如荧光、放射性或离子电流的变化测量蛋白质的活性。
“膜片钳测定”在本文中以最广泛的意义使用，指代用于评估离子在例如不同溶液条件下穿过含有离子通道蛋白诸如NaX或NaV的细胞膜的小膜片的运动的变化的测定。
如本文所用，术语“肽”指代通过肽键连接的五十个或更少氨基酸的链，包括2至50、2至15、2至10、2至8或6至14个氨基酸的氨基酸链。
如本文所用，术语“小分子”指代具有50道尔顿至2500道尔顿的分子量的有机分子。
如本文所用，术语“大环”或“大环分子”指代环状大分子或大分子的大分子环状部分。大环的大小在500道尔顿至7500道尔顿范围内。在本文的一些情况下，大环是环状肽或肽衍生物。
如本文所用，术语“结合片段”指代预期直接与靶标蛋白接触的较大分子的一部分，诸如小分子、肽或抗体。结合片段可用于高通量筛选。
在本公开中，“结合(binds)”或“结合(binding)”或“特异性结合”及类似术语，当指代“结合”至蛋白质诸如NaX或NaV蛋白的分子时，例如，意指结合亲和力足够强烈，使得结合对成员之间的相互作用不能归因于随机分子结合(即，“非特异性结合”)。因此，该结合是选择性的或特异性的。
如本文所用，术语“竞争测定”指代一种测定，其中被测试分子阻止或抑制参考分子与共同靶标的特异性结合。
其他定义酌情包含在以下部分中。
II.突变型NAx蛋白
在一些实施例中，本发明包含突变型人NaX离子通道蛋白。
人NaX是电压门控钠通道蛋白家族的成员，且也称为Nav2.1或scn7a。例如，关于该蛋白质的基因组和氨基酸序列的信息可以在UniProt数据库中在登录号Q01118下找到。在一些实施例中，人NaX的氨基酸序列，包括该信号序列，是SEQ ID NO：1的氨基酸序列。据报道，人NaX蛋白在中枢神经系统并且还在皮肤和其他上皮细胞中作为钠传感器。人NaX和NaV蛋白包含四个结构域(DI、DII、DIII和DIV)，每个结构域包含六个跨膜α螺旋(S1、S2、S3、v4、S5和S6)。这些结构域内存在其他区域，如下表1中所述，每个区域具有对应的氨基酸残基位置。例如，每个结构域还包含“电压传感器结构域”或“VSD”(例如，VSD1、VSD2、VSD3、VSD4)或“电压传感器样结构域”或“VSLD””(例如，VSLD1、VSLD2、VLSD3、VSLD4)，包含S1-S4螺旋及其中间的接头或环；α螺旋之间的特异性接头或环，其出现在蛋白质的细胞外或细胞内表面上，并且其包括例如S4-S5接头和在S5与S6之间的成孔选择性过滤环；以及在四个结构域之间的接头。SEQ ID NO：1的人NaX的特异性螺旋、接头、环和其他区域如下所示：
本公开包括例如突变型人NaX，其中至少一个氨基酸残基经野生型哺乳动物或人NaV的至少一个对应残基取代。在一些情况下，野生型人NaX的区域或完整结构域经哺乳动物或人NaV的等效区域或结构域取代，例如，DIII或DIII/DIV接头中的一些或全部。在此类情况下，突变型人NaX在本文中可替代地指代为“嵌合型人NaX”或“嵌合型NaX”或“嵌合体”，因为它含有来自不同蛋白质、哺乳动物或人NaV的氨基酸序列片段。在一些情况下，经取代的残基或区域来自哺乳动物或人Nav1.1、Nav1.2、Nav1.3、Nav1.4、Nav1.5、Nav1.6、Nav1.7、Nav1.8或Nav1.9。在一些情况下，经取代的残基或区域来自人NaV，诸如来自人Nav1.1、Nav1.2、Nav1.3、Nav1.4、Nav1.5、Nav1.6、Nav1.7、Nav1.8或Nav1.9，例如，来自人Nav1.7。
在一些实施例中，人NaX的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换。在一些情况下，突变型人NaX包含来自人或哺乳动物Nav的DIII S1-S6区域中的全部或部分，任选地其中经置换的人NaX的DIII结构域的至少一部分包含：(a)DIII(残基920-1200)；(b)DiII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)；(c)DiII VSD3、S4-S5接头和S5(残基920-1078)；(d)DIII和DII-DIII接头(残基733-1200)；(e)DIII和DIII-DIV接头(920-1237)；(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。在一些情况下，突变型人NaX包含来自人或哺乳动物Nav的DIII S1-S6区域中的全部或部分，任选地其中经置换的人NaX的DIII结构域的至少一部分由以下组成：(a)DIII(残基920-1200)；(b)DIII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)；(c)DIII VSD3、S4-S5接头和S5(残基920-1078)；(d)DIII和DII-DIII接头(残基733-1200)；(e)DIII和DIII-DIV接头(920-1237)；(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。在一些情况下，人NaX的经人或哺乳动物NaV的DIII结构域的对应部分置换的DIII结构域的至少一部分不包含S5。在一些情况下，人NaX的经人或哺乳动物NaV的DIII结构域的对应部分置换的DIII结构域的至少一部分不包含S6。在一些情况下，经人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的至少一部分不包含S1、S2、S3、S4和/或S4-S5接头中的任一者。在一些情况下，人人NaX的DIII结构域的至少一部分经哺乳动物Nav1.1、哺乳动物Nav1.2、哺乳动物Nav1.3、哺乳动物Nav1.4、哺乳动物Nav1.5、哺乳动物Nav1.6、哺乳动物Nav1.7、哺乳动物Nav1.8、哺乳动物Nav1.9、人Nav1.1、人Nav1.2、人Nav1.3、人Nav1.4、人Nav1.5、人Nav1.6、人Nav1.7、人Nav1.8或人Nav1.9的对应部分置换。例如，在一些情况下，人Nax的DIII结构域的至少一部分经人Nav1.7的对应部分置换。在一些情况下，突变型人人NaX是嵌合体构建体2、嵌合体构建体3、嵌合体构建体7、嵌合体构建体9、嵌合体构建体11、嵌合体构建体15或嵌合体构建体19，或包含SEQ ID No：3、4、8、10、12、16或20中的任一者的氨基酸序列。(参见下表4和图18。)
在其他情况下，突变型人人NaX具有以极性或带电残基或甘氨酸或脯氨酸残基对其S6区段中的一个、两个、三个或所有四个S6区段上的氨基酸残基的至少一个取代。在一些情况下，经取代的残基是极性的或荷电的。示例性极性氨基酸残基包括丝氨酸、苏氨酸、酪氨酸、天冬酰胺、谷氨酰胺、组氨酸。示例性带电氨基酸残基包括天冬氨酸、谷氨酸、赖氨酸和精氨酸。这些氨基酸取代可以跨越S6区段中的一个、两个、三个或所有四个S6区段的细胞内到中间跨膜区域(即，最多是跨膜双层的半程)。对用于设计突变的NaX跨膜区段的预测可以基于可从蛋白质数据库(PDB)获得的可用实验结构和与NaV通道(例如ClustalW)的基于标准结构的多序列比对作出。图20描述人NaX中氨基酸取代的潜在位置，在DI、DII、DIII和DIV中的每一者的S6处，以及它们与其他人人NaV蛋白中的对应残基的比对。如每个序列右侧的虚线框所示，可以在残基L390、F724、I1189或I1492中的任何一者处或在任一方向上最多7个残基的任何残基处进行氨基酸取代，例如，在残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。因此，在一些实施例中，D1的S6、DII的S6、DIII的S6和DIV的S6中的至少一者含有以甘氨酸、脯氨酸、极性或带电残基对上述范围内的氨基酸残基的至少一个取代。
在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的一个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的两个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6αα螺旋中的三个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的所有四个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的至少三个S6α螺旋上的氨基酸残基的一个取代。在一些情况下，突变型人NaX包含在SEQ ID NO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含在SEQ ID NO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人Nax包含在SEQ ID NO：1的以下区段中的至少三个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX在SEQ ID NO：1的以下区段中的每个区段内包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对在残基L390、F724、I1189和I1492中的两者或更多者处的氨基酸残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对残基L390、F724、I1189和I1492中的三者处的氨基酸残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基L390、F724和I1189的取代或对氨基酸残基F724、I1189和I1492的取代。在一些情况下，相较于野生型人Nax，突变型人NaX包含取代F724Q、I1189T和I1492T。在一些情况下，相较于野生型人NaX，该人NaX包含取代L390E、I1189E和I1492E。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对残基L390、F724、I1189和I1492中的所有四者处的氨基酸残基的取代。
在一些实施例中，突变型人NaX蛋白在离子通道测定中具有活性。例如，在一些情况下，离子通道测定涉及在非洲爪蟾卵母细胞中表达该突变蛋白。在一些此类情况下，表达该构建体的非洲爪蟾卵母细胞在从0 mV的保持电位开始在+80mV或-100mV引发的电流振幅显著高于来自表达人NaX野生型的卵母细胞的电流振幅。(参见图18针对几种活性和非活性突变型人NaX蛋白构建体的说明，其中星号表示统计学显著性)。
III.使用突变型人NAx蛋白的筛选方法
本公开尤其还涵盖鉴定作为人NaX调节剂的分子的方法，该方法包括例如在离子通道测定及/或结合测定中筛选针对突变型人NaX的潜在调节剂。
在一些实施例中，人NaX的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换。在一些情况下，突变型人NaX包含来自人或哺乳动物NaV的DIII S1-S6区域中的全部或部分，任选地其中经置换的人NaX的DIII结构域的至少一部分包含：(a)DIII(残基920-1200)；(b)DIII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)；(c)DIII VSD3、S4-S5接头和S5(残基920-1078)；(d)DIII和DII-DIII接头(残基733-1200)；(e)DIII和DIII-DIV接头(920-1237)；(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。在一些情况下，突变型人NaX包含来自人或哺乳动物NaV的DIII S1-S6区域中的全部或部分，任选地其中经置换的人NaX的DIII结构域的至少一部分由以下组成：(a)DIII(残基920-1200)；(b)DIII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)；(c)DIII VSD3、S4-S5接头和S5(残基920-1078)；(d)DIII和DII-DIII接头(残基733-1200)；(e)DIII和DIII-DIV接头(920-1237)；(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。在一些情况下，人NaX的经人或哺乳动物Nav的DIII结构域的对应部分置换的DIII结构域的至少一部分不包含S5。在一些情况下，人NaX的经人或哺乳动物NaV的DIII结构域的对应部分置换的DIII结构域的至少一部分不包含S6。在一些情况下，经人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的至少一部分不包含S1、S2、S3、S4和/或S4-S5接头中的任一者。在一些情况下，人Nax的DIII结构域的至少一部分经哺乳动物Nav1.1、哺乳动物Nav1.2、哺乳动物Nav1.3、哺乳动物Nav1.4、哺乳动物Nav1.5、哺乳动物Nav1.6、哺乳动物Nav1.7、哺乳动物Nav1.8、哺乳动物Nav1.9、人Nav1.1、人Nav1.2、人Nav1.3、人Nav1.4、人Nav1.5、人Nav1.6、人Nav1.7、人Nav1.8或人Nav1.9的对应部分置换。例如，在一些情况下，人NaX的DIII结构域的至少一部分经Nav1.7的对应部分置换。在一些情况下，突变型人NaX是嵌合体构建体2、嵌合体构建体3、嵌合体构建体7、嵌合体构建体9、嵌合体构建体11、嵌合体构建体15或嵌合体构建体19，或包含SEQ ID No：3、4、8、10、12、16或20中的任一者的氨基酸序列。(参见下表4和图18。)
本公开还涵盖鉴定人NaX离子通道蛋白(NaX)调节剂的方法，该方法包括：(a)提供突变型人NaX，其包含以甘氨酸、脯氨酸或极性或带电残基对人NaX中的两个、三个或四个S6α螺旋中的每个S6α螺旋上的至少一个残基的取代；(b)在潜在调节剂的存在下对突变型人NaX进行离子通道测定；(c)如果在潜在调节剂的存在下测定中的突变型人NaX的活性高于或低于在潜在调节剂不存在下的活性，则将该潜在调节剂鉴定为人NaX调节剂。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的两个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人Nax包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的一个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人Nax包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的两个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的三个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的所有四个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的至少三个S6α螺旋上的氨基酸残基的一个取代。在一些情况下，突变型人Nax包含在SEQ ID NO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含在SEQ IDNO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含在SEQ ID NO：1的以下区段中的至少三个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人Nax在SEQ ID NO：1的以下区段中的每个区段内包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对在残基L390、F724、I1189和I1492中的两者或更多者处的氨基酸残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对残基L390、F724、I1189和I1492中的三者处的氨基酸残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基L390、F724和I1189的取代或对氨基酸残基F724、I1189和I1492的取代。在一些情况下，相较于野生型人NaX，突变型人NaX包含取代F724Q、I1189T和I1492T。在一些情况下，相较于野生型人NaX，该人NaX包含取代L390E、I1189E和I1492E。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对残基L390、F724、I1189和I1492中的所有四者处的氨基酸残基的取代。
本公开进一步包括确定调节人离子通道蛋白(例如，人NaV蛋白)活性的测试分子是否也调节人NaX离子通道(NaX)蛋白的活性的方法，该方法包括：(a)提供突变型人NaX，其中人NaX的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换；(b)在测试分子的存在下对该突变型人NaX进行离子通道测定；以及(c)如果在测试分子的存在下该测定中的突变型人NaX的活性高于或低于在测试分子不存在下的活性，则将该测试分子确定为突变型人NaX调节剂；以及任选地，(d)如果根据(c)部分，测试分子不是人NaX调节剂，则选择该测试分子进行额外筛选。
本公开额外包括确定调节人离子通道蛋白(诸如人Nav蛋白)活性的测试分子是否也调节人NaX离子通道(NaX)蛋白的活性的方法，该方法包括：(a)确定测试分子调节人离子通道蛋白的活性；(b)提供突变型人Nax，其中人NaX的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换；(c)在测试分子的存在下对该突变型人Nax进行离子通道测定；和(d)如果在测试分子的存在下该测定中的突变型人Nax的活性高于或低于在测试分子的不存在下的活性，则将该测试分子确定为突变型人NaX调节剂；以及任选地，(e)如果根据(d)部分，测试分子不是人Nax调节剂，则选择该测试分子进行额外筛选。
在一些实施例中，人Nax的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换。在一些情况下，突变型人NaX包含来自人或哺乳动物NaV的DIII S1-S6区域中的全部或部分，任选地其中经置换的人NaX的DIII结构域的至少一部分包含：(a)DIII(残基920-1200)；(b)DIII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)；(c)DIII VSD3、S4-S5接头和S5(残基920-1078)；(d)DIII和DII-DIII接头(残基733-1200)；(e)DIII和DIII-DIV接头(920-1237)；(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。在一些情况下，突变型人Nax包含来自人或哺乳动物NaV的DIII S1-S6区域中的全部或部分，任选地其中经置换的人NaX的DIII结构域的至少一部分由以下组成：(a)DIII(残基920-1200)；(b)DIII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)；(c)DIII VSD3、S4-S5接头和S5(残基920-1078)；(d)DIII和DII-DIII接头(残基733-1200)；(e)DIII和DIII-DIV接头(920-1237)；(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。在一些情况下，人NaX的经人或哺乳动物Nav的DIII结构域的对应部分置换的DIII结构域的至少一部分不包含S5。在一些情况下，人NaX的经人或哺乳动物NaV的DIII结构域的对应部分置换的DIII结构域的至少一部分不包含S6。在一些情况下，经人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的至少一部分不包含S1、S2、S3、S4和/或S4-S5接头中的任一者。在一些情况下，人NaX的DIII结构域的至少一部分经哺乳动物Nav1.1、哺乳动物Nav1.2、哺乳动物Nav1.3、哺乳动物Nav1.4、哺乳动物Nav1.5、哺乳动物Nav1.6、哺乳动物Nav1.7、哺乳动物Nav1.8、哺乳动物Nav1.9、人Nav1.1、人Nav1.2、人Nav1.3、人Nav1.4、人Nav1.5、人Nav1.6、人Nav1.7、人Nav1.8或人Nav1.9的对应部分置换。例如，在一些情况下，ANaX的DIII结构域的至少一部分经Nav1.7的对应部分置换。在一些情况下，突变型人NaX是嵌合体构建体2、嵌合体构建体3、嵌合体构建体7、嵌合体构建体9、嵌合体构建体11、嵌合体构建体15或嵌合体构建体19，或包含SEQ ID No：3、4、8、10、12、16或20中的任一者的氨基酸序列。(参见下表4和图18。)
本公开还包括确定调节人离子通道蛋白活性的测试分子是否调节人NaX离子通道(NaX)蛋白的活性的方法，该方法包括：(a)提供突变型人NaX，其包含以甘氨酸、脯氨酸或极性或带电残基对人NaX的两个、三个或四个S6α螺旋中的每个S6α螺旋上的至少一个残基的取代；(b)在测试分子的存在下对突变型人Nax进行离子通道测定；(c)如果在测试分子的存在下该测定中的突变型人Nax的活性高于或低于在潜在测试分子不存在下的活性，则将该测试分子确定为人Nax调节剂；以及任选地，(d)如果根据(c)部分，测试分子不是人NaX调节剂，则选择该测试分子进行额外筛选。
本公开进一步涵盖确定调节人离子通道蛋白活性的测试分子是否也调节人NaX离子通道(Nax)蛋白的活性的方法，该方法包括：(a)确定测试分子调节人离子通道蛋白的活性；(b)提供突变型人NaX，其包含以甘氨酸、脯氨酸或极性或带电残基对人NaX的两个、三个或四个S6α螺旋中的每个S6α螺旋上的至少一个残基的取代；(c)在测试分子的存在下对突变型人Nax进行离子通道测定；(d)如果在测试分子的存在下该测定中的突变型人Nax的活性高于或低于在潜在测试分子不存在下的活性，则将该测试分子确定为人NaX调节剂；以及任选地，(e)如果根据(d)部分，测试分子不是人NaX调节剂，则选择该测试分子进行额外筛选。
在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的一个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的两个S6αα螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的三个S6αα螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的所有四个S6α螺旋上的至少一个残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对四个S6α螺旋中的至少三个S6α螺旋上的氨基酸残基的一个取代。在一些情况下，突变型人NaX包含在SEQ ID NO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含在SEQ ID NO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含在SEQ ID NO：1的以下区段中的至少三个区段内以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX在SEQ ID NO：1的以下区段中的每个区段内包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对在残基L390、F724、I1189和I1492中的两者或更多者处的氨基酸残基的取代。在一些情况下，突变型人NaX包含以甘氨酸、脯氨酸或极性或带电残基对残基L390、F724、I1189和I1492中的三者处的氨基酸残基的取代。在一些情况下，突变型人Nax包含以甘氨酸、脯氨酸或极性或带电残基对氨基酸残基L390、F724和I1189的取代或对氨基酸残基F724、I1189和I1492的取代。在一些情况下，相较于野生型人NaX，突变型人NaX包含取代F724Q、I1189T和I1492T。在一些情况下，相较于野生型人NaX，该人NaX包含取代L390E、I1189E和I1492E。在一些情况下，突变型人Nax包含以甘氨酸、脯氨酸或极性或带电残基对残基L390、F724、I1189和I1492中的所有四者处的氨基酸残基的取代。
在确定已知或发现调节人离子通道蛋白活性的测试分子是否也调节人NaX离子通道(NaX)蛋白活性的某些方法中，这些方法可用作反选择。例如，如果实验者希望确保特定NaV蛋白的调节剂对该NaV蛋白和/或相关NaV蛋白具有特异性，则实验者可以进行本文的方法以确认测试分子不是该突变型人NaX的调节剂。
在任何上述方法中，离子通道测定可以是，例如，用于检测突变型NaX蛋白作为离子通道的活性的任何合适测定。示例包括但不限于膜片钳测定，包括自动化膜片钳测定、离子通量测定以及离子敏感或电压敏感染料测定。示例性测定在例如H.Yu等人，“highthroughput screening technologies for ion channels,”Acta PhaFm.Sinica 37：34-43(2016)中描述，且材料可从制造商获得。在离子通量测定中，例如，放射性同位素，诸如钠22(22Na+)，可用于追踪钠离子或其他离子的细胞流入和流出。另一种类型的测定是电压敏感或离子敏感染料测定。在电压敏感染料测定中，使用荧光共振能量转移(FRET)测量跨过包含离子通道蛋白的膜的电压变化，例如，使用染料诸如氧喏(oxonol)衍生物诸如双-(1，3-二丁基巴比妥酸)三甲基氧喏(DiBAC4)或FMP。在一些情况下，FRET染料可能会定位或束缚至膜。离子敏感染料测定可以使用根据离子浓度显示信号差异的染料。一个示例是钠指示染料SBFI。在其他实施例中，可以在筛选方法中使用膜片钳测定。在一些实施例中，可以使用自动化钳测定。用于进行膜片钳测定的示例性平台和仪器由多家制造商售卖。示例性平台测定包括IonWorksTM平台测定、PatchXpressTM和IonFluxTM(Molecular Devices)、QpatchTM HT/HTX(Sophion)、以及PatchlinerTM和SynchroPatchTM(Nanion Technologies)。
在任何上述方法中，在该方法中鉴定的潜在调节剂调节突变型人Nax的活性，例如，具有低于或高于野生型人Nax的活性。在一些情况下，潜在调节剂降低突变型人NaX的活性，诸如具有低于野生型人NaX的活性。在一些情况下，可以将活性降低至少25％、至少30％、至少40％、至少50％、至少60％、至少70％、至少75％、至少80％或至少90％。在一些情况下，突变型人NaX在调节剂的存在下的活性可以是，例如，突变型人Nax在调节剂不存在下的活性的1-90％、10-90％、1-10％、1-20％、25-90％、50-90％、25-75％、40-80％或50-75％。在一些情况下，在该测定中鉴定的潜在调节剂以10nM至500μM，50nM至500μM，10nM至50μM，100nM至500μM，100nM至50μM，1-500μM，1-50μM，10-500μM或50-250μM的半最大浓度使该测定中的突变型人Nax的活性降低。在其他情况下，该测定中鉴定的潜在调节剂使该测定中的突变型人NaX的活性增加，诸如具有高于野生型人NaX的活性。在一些情况下，可以使活性增加至少10％、至少20％、至少25％、至少30％、至少40％、至少50％、至少60％、至少70％、至少75％、至少80％、或至少100％(即，两倍)、至多100％、或至多三倍。在一些情况下，突变型人NaX在调节剂的存在下的活性可以比突变型人Nax在调节剂不存在下的活性高10-100％、10-80％、20-80％、25-100％、25-75％、50-100％、两至三倍或100-200％。
在任何上述方法中，离子通道测定也可以在突变型人NaX的潜在调节剂和经鉴定的调节剂两者的存在下进行。例如，在这种情况下，人们可能会确定所鉴定的调节剂是否与潜在调节剂竞争以改变蛋白质的活性。在一些此类情况下，突变型人NaX的经鉴定的调节剂是河豚毒素(TTX)、奎尼丁(quinidine)、氟卡尼(flecainide)、丁卡因(tetracaine)或利多卡因(lidocaine)中的一者或多者。
上述任何方法还可进一步包含确定在该测定中鉴定的潜在调节剂与突变型人NaX和野生型人NaX中任一者或两者的结合亲和性，以及在嵌合型人NaX的情况下，与经取代的氨基酸自其衍生的哺乳动物或人Nav蛋白的结合亲和性。例如，ELISA测定可用于确定结合亲和力，例如，使用在基质诸如固体表面(诸如珠、板等)上的脂质稳定形式的突变型人NaX蛋白。珠可以具有任何形状，诸如薄片或碎片、球体、丸粒等。在一些实施例中，此类珠是经链霉亲和素包被的珠、经亲和素包被的珠或经去糖基化的亲和素包被的珠。在一些实施例中，此类珠是磁珠。在一些情况下，潜在调节剂以10μM或更少、10μM至50nM、10μM至500nM、1μM或更少、1μM至50nM、或100nM或更少的EC50或IC50，与突变型人NaX和野生型人NaX中的任一者或两者结合。
在一些实施例中，可以对在上述筛选中选择的分子进行进一步实验，例如，以确定分子的其他生物活性。例如，可以使用进一步的测定来确定该分子是否也结合其他离子通道蛋白，诸如人或哺乳动物NaV蛋白，从而确定该分子作为离子通道调节剂或结合剂的特异性。例如，在使用嵌合型人NaX的方法中，可以进行进一步测定以确定该分子是否也调节自其衍生出突变型NaX的嵌合部分的NaV蛋白的活性，或者该分子是否与该NaV蛋白结合。
IV.用于筛选方法的测试分子
在任何上述方法中，潜在调节剂(即测试分子)可以是肽或大环或抗体。例如，在一些情况下，潜在调节剂是小分子。本公开还涉及通过本文所述的方法鉴定的人NaX调节剂，其可以任选地为肽、大环、小分子或抗体。
在一些实施例中，待测试的潜在调节剂分子是肽。在一些实施例中，肽是6-14-mer肽，诸如6-12-mer、6-10-mer、6-8-mer、8-12-mer、8-10-mer等。参见载玻片10。在一些实施例中，肽是14-mer。参见载玻片15-16。在一些实施例中，肽是6-10mer。在一些实施例中，肽是8-10mer。在一些实施例中，肽是6-8mer。在一些实施例中，肽是8-mer。参见载玻片21-22。在一些实施例中，肽是3-40-mer、3-20-mer、4-16-mer、4-14-mer或6-14-mer，诸如3-mer、4-mer、5-mer、6-mer、7-mer、8-mer、9-mer、10-mer、11-mer、12-mer、13-mer、14-mer、15-mer、16-mer、17-mer、18-mer、19-mer、20-mer、21-mer、22-mer、23-mer、24-mer、25-mer、26-mer、27-mer、28-mer、29-mer、30-mer、31-mer、32-mer、33-mer、34-mer、35-mer、36-mer、37-mer、38-mer、39-mer或40-mer。
在一些实施例中，肽是大环。在一些实施例中，大环是6-14-mer大环，诸如6-12-mer、6-10-mer、6-8-mer、8-12-mer、8-10-mer等。参见载玻片10。在一些实施例中，大环是14-mer大环。参见载玻片15-16。在一些实施例中，大环是6-10-mer大环。在一些实施例中，大环是8-10-mer大环。在一些实施例中，大环是6-8-mer大环。在一些实施例中，大环是8-mer大环。参见载玻片21-22。在一些实施例中，大环是3-40-mer、3-20-mer、4-16-mer、4-14-mer或6-14-mer，诸如3-mer、4-mer、5-mer、6-mer、7-mer、8-mer、9-mer、10-mer、11-mer、12-mer、13-mer、14-mer、15-mer、16-mer、17-mer、18-mer、19-mer、20-mer、21-mer、22-mer、23-mer、24-mer、25-mer、26-mer、27-mer、28-mer、29-mer、30-mer、31-mer、32-mer、33-mer、34-mer、35-mer、36-mer、37-mer、38-mer、39-mer或40-mer大环。在一些实施例中，大环具有至少一个亲脂侧链和至少一个带正电荷的侧链。
在一些实施例中，本文筛选中待测试的分子是小分子。在一些实施例中，待测试的分子是抗体，其不仅可以包括任何IgG、IgM、IgA、IgD和IgE的全长抗体，而且可以包括抗体的抗原结合片段，诸如Fv，Fab′、(Fab′)2、scFv等、纳米抗体、单链抗体、双特异性或多特异性抗体。
在一些实施例中，待测试的分子是肽的结合片段、小分子的结合片段或抗体的结合片段(例如，抗原结合片段)。
V.分子复合物
在一些实施例中，本公开包括分子复合物，该分子复合物包含与分子(诸如潜在调节剂分子，诸如肽、小分子、抗体，或者肽、小分子或抗体的结合片段)结合的如本文所述的突变型人NaX。
在一些实施例中，该分子是肽。在一些实施例中，肽是6-14-mer肽，诸如6-12-mer、6-10-mer、6-8-mer、8-12-mer、8-10-mer等。参见载玻片10。在一些实施例中，肽是14-mer。参见载玻片15-16。在一些实施例中，肽是6-10mer。在一些实施例中，肽是8-10mer。在一些实施例中，肽是6-8mer。在一些实施例中，肽是8-mer。参见载玻片21-22。在一些实施例中，肽是3-40-mer、3-20-mer、4-16-mer、4-14-mer或6-14-mer，诸如3-mer、4-mer、5-mer、6-mer、7-mer、8-mer、9-mer、10-mer、11-mer、12-mer、13-mer、14-mer、15-mer、16-mer、17-mer、18-mer、19-mer、20-mer、21-mer、22-mer、23-mer、24-mer、25-mer、26-mer、27-mer、28-mer、29-mer、30-mer、31-mer、32-mer、33-mer、34-mer、35-mer、36-mer、37-mer、38-mer、39-mer或40-mer。
在一些实施例中，肽是大环。在一些实施例中，大环是6-14-mer大环，诸如6-12-mer、6-10-mer、6-8-mer、8-12-mer、8-10-mer等。参见载玻片10。在一些实施例中，大环是14-mer大环。参见载玻片15-16。在一些实施例中，大环是6-10-mer大环。在一些实施例中，大环是8-10-mer大环。在一些实施例中，大环是6-8-mer大环。在一些实施例中，大环是8-mer大环。参见载玻片21-22。在一些实施例中，大环是3-40-mer、3-20-mer、4-16-mer、4-14-mer或6-14-mer，诸如3-mer、4-mer、5-mer、6-mer、7-mer、8-mer、9-mer、10-mer、11-mer、12-mer、13-mer、14-mer、15-mer、16-mer、17-mer、18-mer、19-mer、20-mer、21-mer、22-mer、23-mer、24-mer、25-mer、26-mer、27-mer、28-mer、29-mer、30-mer、31-mer、32-mer、33-mer、34-mer、35-mer、36-mer、37-mer、38-mer、39-mer或40-mer大环。在一些实施例中，大环具有至少一个亲脂侧链和至少一个带正电荷的侧链。
在一些实施例中，复合物中的该分子是小分子。在一些实施例中，该分子是抗体，其不仅可以包括任何IgG、IgM、IgA、IgD和IgE的全长抗体，而且可以包括抗体的抗原结合片段，诸如Fv，Fab′、(Fab′)2、scFv等、纳米抗体、单链抗体、双特异性或多特异性抗体。
在一些实施例中，该分子是肽的结合片段、小分子的结合片段或抗体的结合片段(例如，抗原结合片段)。
VII.试剂盒
本公开还包括包含与本文的筛选方法相关的试剂的试剂盒。在一些情况下，试剂盒包括一种或多种如本文所述的突变型人NaX。在一些情况下，试剂盒包括用于本文的筛选方法的试剂，具有或不具有特定的突变型人Nax。在某些情况下，试剂盒包括超过一种类型的突变型人NaX。在一些情况下，试剂盒还包括至少一种哺乳动物或人NaV和/或野生型人NaX，例如，作为对照。
在一些实施例中，本文的试剂盒可包括一种或多种用于进行离子通道测定的试剂。在一些情况下，试剂盒可包括突变型人NaX的特定调节剂，例如，作为阳性对照。本文的试剂盒还可包括经鉴定为不调节突变型人NaX的阴性对照。在使用试剂盒来确定分子是否与突变蛋白结合的情况下，试剂盒可以包括与基质粒子(诸如珠)或另一种类型的基质(诸如板)附接的突变型人NaX。珠可以具有任何形状，诸如薄片或碎片、球体、丸粒等。在一些实施例中，此类珠是经链霉亲和素包被的珠、经亲和素包被的珠或经去糖基化的亲和素包被的珠。在一些实施例中，此类珠是磁珠。突变型人NaX可以或可以不预先附接至基质。在一些实施例中，包括试剂以促进蛋白质附接至基质，诸如通过生物素-链霉亲和素或类似系统。
在一些实施例中，试剂盒可包括的筛选方法相关的试剂，诸如进行本文方法中所述的离子通道测定所需的一些或所有试剂。在一些情况下，还可包括一种或多种对照试剂，诸如蛋白质的调节剂。
在一些实施例中，试剂盒可包括测试分子或测试分子文库，诸如肽、小分子和/或抗体。在一些实施例中，试剂盒中的肽可以是大环。在一些实施例中，试剂盒可包括测试分子，该测试分子是肽、小分子或抗体的结合片段。
在一些实施例中，试剂盒还可包括使用说明。
实例
以下是本公开的方法和组合物的实例。应理解，这些实例并非意在限制本公开，而仅用于举例说明，并且在给出以上提供的一般描述的情况下可以实施各种其他实施例。
实例1-材料和方法
实例1.1-实验模型和受试者细节
细胞系Expi293F是经转化并用于悬浮生长(Thermo Fisher Scientific)的人胚肾细胞系(雌性)，并用于蛋白质表达。Expi293F细胞在SMM 293T-I培养基中在37℃和5％CO2下培养。验证Expi293F细胞并进行支原体(Mycoplasma)污染测试。
实例1.2-蛋白质表达和纯化
在其N末端含有串联2xStrepII和2xFLAG标签的全长人NaX以及未标记的全长人β3在CMV启动子的控制下分别克隆到pRK载体中。两种构建体均使用PEI转染到Expi293细胞中，并在SMM 293T-I培养基中在5％CO2和37℃下培养48小时。通过在800g离心10分钟收获细胞，并通过杜恩斯(dounce)均质化在缓冲液A(25mM ADA pH 6.0、200mM NaCl、1mMPMSF、1μg/mL全能核酸酶(benzonase)和1x Roche蛋白酶抑制剂混合物)中重悬和裂解。为了溶解细胞膜，将葡糖-薯蓣皂苷元(glyco-diosgenin(GDN))和胆固醇半琥珀酸盐(CHS)添加到样品中，最终浓度分别为2％和0.3％，并在4℃轻轻搅拌样品两小时。然后通过在4℃以125，000g超速离心1小时收集不溶物质。将上清液施加至已在缓冲液B(25mM ADA pH 6.0、200mMNaCl、0.01％GDN)中预平衡的抗FLAG M2琼脂糖树脂上，并在4℃分批结合1小时。然后将样品施加至重力管柱上，并且所收集的树脂用10CV缓冲液B洗涤，然后用补充有5mM ATP和10mM MgCl2的10CV洗涤。用补充有300μg/mL FLAG肽的5CV缓冲液B洗脱蛋白质。将洗脱液直接施加至Strep-Tactin XT Superflow高容量树脂上，该树脂已在缓冲液B中预平衡并分批结合三小时，然后用10CV缓冲液B洗涤，然后用补充有50mM生物素的5CV进行样品洗脱。对于纳米盘并入，使用Amicon Ultra离心过滤装置(100kDa MWCO)将样品浓缩至15μM。对于洗涤剂(GDN)中的结构分析，将洗脱液浓缩至100μL，然后施加至已在缓冲液B中预平衡的Superose 6 3.2/300管柱。
实例1.3-质谱分析
在尺寸排阻色谱之后，用8M胍(1：1v/v)使10μg的含NaX的峰级分(在分别与β1-或β3-亚基共表达后)变性，用1M二硫苏糖醇(DTT)(Sigma-Aldrich，St Louis，MO)还原至最终浓度为100mM DTT，并在95℃培育10分钟，然后离心。将10μL的1M Tris，pH 8添加到溶液中，用水将胍稀释至2M最终浓度。用2μL的PNGaseF(15000U，New England Biolabs)进行去糖基化，然后在37℃培育过夜。在60℃用10mM DTT进一步还原样品(15分钟)，然后在室温用20mM碘乙酰胺进行烷基化。将蛋白质用0.2μg胰蛋白酶(Promega)或胰凝乳蛋白酶在50mM碳酸氢铵，pH 8中在37℃消化过夜。用甲酸淬灭消化物，并将上清液在C18 PhyTips(PhyNexus)上脱盐，冻干，在含有2％乙腈的0.1％甲酸中重构，未经进一步处理即通过反相纳米LC/MS/MS在与Elite Orbitrap质谱仪(ThermoFisher Scientific)串接的Waters NanoAcquityHPLC系统(Waters Corp.)上分析。将肽加载至SymmetryC18管柱(1.7mm BEH-130，0.1x100mm，Waters)上，并且用从2％至25％溶剂B(0.1％甲酸，98％乙腈)的60分钟梯度以1μL/min流速分离。肽直接洗脱到喷雾电压为1.2kV的质谱仪中。通过FT获取完整MS数据，范围为350-1250m/z且分辨率为60,000。通过2-Da分离窗口自全MS扫描选择MS/MS丰度最高的离子。
使用具有胰蛋白酶或胰凝乳蛋白酶特异性的Mascot算法(Matrix Sciences)搜索获取的串联MS谱。搜索标准包括50ppm的完整MS容差、0.5Da的MS/MS容差，其中甲硫氨酸氧化(+15.9949Da)作为可变修饰，且半胱氨酸的胺基甲酰甲基化(+57.0215Da)作为静态修饰。针对重叠在Uniprot哺乳动物数据库上的内部构建体的特异性序列(包括反向蛋白质序列)搜索数据。肽分配首先在肽层次过滤至2％的错误发现率(FDR)，然后在蛋白质层次过滤至2％的FDR。
实例1.4-将β3-NaX重建至脂质纳米盘中
为了重建至纳米盘中，将多个50μL样品等分试样分别施加至具有200倍摩尔过量的POPC：POPE：POPG脂质混合物(3：1：1比率，在超音浴中以10mg/mL溶解在含有50mM HEPESpH 7.5、100mM NaCl、5mM MgCl2和1％CHAPS的缓冲液中)的2mL管中，并在4℃培育30分钟。然后将膜支架蛋白MSP1E3D1(Sigma)以4倍摩尔过量施加至样品，并在4℃再培育30分钟。然后将样品稀释至1.5mL，并将Bio-Beads施加至最终浓度为0.25mg/mL。然后在去除Bio-Bead之前，将样品在4℃突变过夜。然后将样品汇集，并使其通过已在缓冲液C(25mM HEPES pH7.0、200mM NaCl)中预平衡的管柱中的1mL Strep-Tactin XT树脂以去除空纳米盘。在用含有50mM生物素的5CV进行样品洗脱之前，用5CV缓冲液C洗涤管柱。然后将样品浓缩至100μL并施加至已在缓冲液C中预平衡的Superose 63.2/300管柱。汇集峰级分并浓缩至3mg/mL。
实例1.5-Cryo-EM样品制备和数据采集
样品在室温与最终浓度为0.05％的EM级戊二醛交联10分钟。通过添加1M Tris pH7.0淬灭交联反应。使用Solarus等离子体清洁器(Gatan，Pleasanton，CA)对Ultrafoil R2/2(200目)cryoEM网格(Quantifoil GMBH)进行等离子体清洁5秒。将三微升最终样品以2.25mg/mL(纳米盘)或1.5mg/mL(GDN)施加至cryoEM网格，并使用Vitribot Mark IV(ThermoFIsher Scientific，Waltham，MA)使用在100％湿度下设置为8的印迹力进行印迹2.5秒，然后投入液态乙烷中。然后，在带有生物量子能量过滤器的在300keV下操作的TitanKrios电子显微镜(ThermoFisher Scientific，Waltham，MA)上，使用K2(纳米盘)或K3(GDN)Summit直接电子检测器(Gatan，Pleasanton，CA)对网格进行成像。以165,000倍放大倍率记录纳米盘的图像，这对应于使用20eV能量狭缝的/像素。图像堆栈含有50张图像，在10秒内以0.2秒的间隔记录，总曝光量为使用20eV能量狭缝以105,000倍放大率(对应于/像素)以超分辨率模式记录GDN样品的图像。图像堆栈含有60张图像，在3秒内以0.05秒的间隔记录，总曝光量为所有数据收集都使用serialEM67完成。
实例1.6-Cryo-EM数据处理
纳米盘样品
使用WARP、RELION和cisTEM软件包的组合来处理cryo-EM数据68-71。对于第一个数据集，在RELION中使用MotionCor272对11,658个视频影像进行了帧移动校正。过滤所得图像以仅保留那些累积运动总和低于250的值的图像，并使用CTFFIND4.173使用光谱的频带拟合反差转换函数(CTF)参数。图像经过滤以仅包括那些所检测的拟合分辨率优于的图像，总共提供了9,988个良好图像以供进一步处理。在WARP68中使用基于深度学习的算法挑选了1,669,107个粒子。粒子在cisTEM中进行两轮2D分类，选择30个最佳类别(76,392个粒子)并输出到RELION进行3D分类。使用在纳米盘(EMD-9024)74中解析的经低通过滤(LPF)的KV1.2重建作为初始3D参考，该重建的纳米盘外的所有密度都已擦除。然后将获得的最佳3D体积用作第二轮3D分类的参考，使用cisTEM中2D分类中更广泛的粒子选择(350,901个粒子)。然后将最佳3D体积及其对应的91,435个粒子重新输入cisTEM，以进行迭代轮次的无需遮蔽的自动细化，并使用经过迭代调整的遮蔽和遮蔽外的LPF(外部权重为0.8)进行手动细化。然后将得到的 图用作cisTEM中最后一轮无遮蔽自动细化和经遮蔽手动细化的参考，使用来自cisTEM中仅单轮2D分类的粒子堆栈，给出最终的图。
洗涤剂(GDN)样品
使用RELION、Gautomatch[K.Zhang，MRC LMB(mrc-lmb.cam.ac.uk/kzhang/)]和cisTEM软件包的组合处理cryo-EM数据。在RELION中使用MotionCor272对10,850个视频影像进行帧移动校正，并合并为/像素。过滤所得图像以仅保留那些累积运动总和低于250的值的图像，并使用CTFFIND4.173使用光谱的频带拟合反差转换函数(CTF)参数。图像经过滤以仅包括那些所检测的拟合分辨率优于的图像，总共提供了10,569个良好图像以供进一步处理。使用Gautomatch自动挑选出总共2,780,663个粒子，使用NaV1.4与β1辅助亚基(EMD-9617)复合的重建的16个均匀投影作为模板。粒子经历在RELION中的单轮2D分类，分为250个类别，并从55个最佳类别中提取包含1,420,422个粒子的粒子堆栈并将其输出到cisTEM。使用在纳米盘中解析的β3-NaX的结构作为第一轮无遮蔽自动细化中的初始参考，随后进行几轮使用迭代遮蔽细化和遮蔽外的LPF(外部权重为0.8)的手动细化。手动细化运行包括散焦细化并使用0.2的得分阈值。迭代轮次的无遮蔽自动细化和遮蔽手动细化产生了分辨率的最终重建。
实例1.7-模型构建和结构分析
使用NaV1.4与β1辅助亚基(PDB 6AGF)28复合的结构作为模板，以使用Phyre2服务器58生成NaX同源模型。将该模型与β1一起刚体对接至cryo-EM图中，用于在Coot59中手动重建，以提供β3-NaX复合物的初始模型。在Phenix60中进行多轮真实空间细化并在Coot中进行手动重建，然后在UCSF ChimeraX61中使用ISOLDE62进行分子动力学辅助的手动细化。在Phenix中进行最终细化后，使用Phenix验证包对模型进行验证。使用PyMol63、UCSFChimera64和UCSF ChimeraX生成结构图。使用Caver3.0来分析通道孔65。使用ClustalOmega66进行序列比对，并使用ESPript 3.067进行渲染。
实例1.8-用于生化和电生理实验的分子生物学
将针对智人进行密码子优化的人NaX、NaX-eGFP-2×FLAG、NaV1.7、NaX-NaV1.7结构域调换嵌合体、ATP1A1、ATP1B1和SAP97的互补DNA(cDNA)克隆到pcDNA3.1/Hygro(+)载体中，用于本研究。将人NaVβ1、NaVβ3、CaVβ1、CaVγ2、CaVα2δδ1、ATP1A1、ATP1B1、ATP1G1和SAP97构建体克隆到pcDNA3.1(+)载体中。使用经定制设计的引物(Eurofins Genomics)、PfuUltraII融合HSDNA聚合酶(Agilent Technologies)进行定点诱变来生成NaX、NaV1.5和NaV1.7突变体(NaX N1142K、QTT(F724Q/I1189T/I1492T)；NaV1.5S1610P/S1618P/A1640P；Nav1.7S211P、ΔL966、S1269F、K1406N和A1596P/S1605P/A1627P)，并通过Sanger DNA测序(EurofinsGenomics)验证DNA序列。为了在非洲爪蟾卵母细胞中表达，使用NotI、BamHI或XbaI限制性内切酶对cDNA进行线性化，然后使用T7 mMessage mMachine试剂盒(Ambion)将其转录为封端RNA。为了在HEK293T和Neuro-2a细胞中表达，使用了用NucleoBond Xtra Midi Plus试剂盒(Macherey-Nagel)纯化的质粒DNA。
实例1.9-双电极电压钳电生理学
如前所述制备卵母细胞68。使用Nanoliter 2010注射器(World PrecisionInstruments)向看起来健康的V-VI期卵母细胞注射2.5-40ng RNA(在5-41nL中)。当排除一种或多种构建体时，添加等体积的无核酸酶水以在不同构建体组合中保持恒定的RNA浓度。经注射的卵母细胞在18℃，140rpm，在补充有50μg/mL庆大霉素和50μg/mL四环素的ND96(以mM为单位：96NaCl、2KCl、1MgCl2、1.8CaCl2、2.5丙酮酸钠、0.5茶碱、5HEPES；用NaOH调节至pH7.4)中保持两到五天。使用Warner OC-725C卵母细胞钳放大器(Wamer Instrument Corp，USA)在室温并在以ND96溶液(以mM为单位：96NaCl、2KCl、1MgCl2、1.8CaCl2、5HEPES，用NaOH调节至pH 7.4)进行的恒定灌注下进行双电极电压钳测量。对于离子选择性实验，外部溶液含有115mM的测试阳离子，如氯化物盐，1.2mM CaCl2、2mM MgCl2、5mM HEPES(用对应氢氧化物调节至pH 7.4)。使用Digidara 1550数字转换器(Moleculardevices；以10kHz采样)和pCLAMP 10软件(Molecular Devices)进行数据采集。使用P-1000火焰/棕色微移液管拉拔器系统(Sutter Instrument)从硼硅酸盐玻璃毛细管(Harvard Apparatus)制备微电极，使其电阻约为0.2-1.0MΩ，并用3M KCl回填。将乌头碱和藜芦定碱(Alomone Labs)溶解在DMSO中制成50mM储备液，并在ND96中进一步稀释至300和100μM用于电生理实验。将其他NaV活化剂(Alomone Labs；图8A-8D)溶解在水中制成储备溶液。离子电流的电压依赖性使用由阶跃组成的程序确定，该阶跃从0mV的保持电位到+80至-100mV的电压范围，持续1s，以20mV为增量。
实例1.10-细胞培养、细胞表面生物素化和蛋白质印记
HEK293T和Neuro-2a细胞(ATCC)如前所述生长并维持68。将大约800,000个HEK293T细胞和250,000个Neuro-2a细胞接种在35mm细胞培养皿中～20小时，然后使用LipoD293版本II(tebu-bio)用1μg cDNA进行瞬时转染。对于生化实验，将NaX-eGFP-2×FLAG和β3的cDNA以1：1的质量比混合。当β3从转染中排除时，添加等量的空载体DNA以保持总cDNA量恒定。在转染后24小时，将经转染的HEK293T细胞用于生化实验。为了诱导分化，Neuro-2a细胞在转染后24小时进行血清饥饿处理(在DMEM中再培养24-30小时)，然后用于生化和电生理实验。如前所述进行细胞表面生物素化和蛋白质印记68，仅稍作修改：1)淬灭步骤在冰上在轻轻搅拌下进行30分钟，2)非洲爪蟾卵母细胞在裂解前用tris缓冲盐水洗涤两次，以及3)由于蛋白质过多，在加载到凝胶上之前，卵母细胞的总裂解物级分在SDS样品缓冲液中按1：5稀释。如68中所述使用抗体，但小鼠抗Na+/K+-ATP酶抗体来自Santa Cruz Biotechnology(sc-21712)。印迹代表至少三个独立实验。
实例1.11-膜片钳电生理学
在膜片钳实验前一小时，将表达NaX-eGFP-2×FLAG的经分化的Neuro-2a细胞接种在玻璃盖玻片上。使用Axopatch 200B放大器(Molecular Devices)、Digidara 1550A数字转换器(Molecular Devices；以10kHz采样)和pCLAMP 10软件(Molecular Devices)在全细胞配置中以-60mV电压钳位细胞(图1E-1F和7F)。用于膜片钳实验的玻璃移液管从硼硅酸盐玻璃毛细管(World Precision Instruments；Kwik-Fil 1.5/1.12；OD/ID)拉出，并经回火抛光至最终电阻在2.0-5.5MΩ之间。细胞外溶液含有(以mM为单位)：140或190NaCl、5KCl、2.5CaCl2、1MgCl2、5HEPES、20葡萄糖(用NaOH调节至pH 7.4)。这些溶液的渗透压值为～302mOsm/L(对于140NaCl)和～395mOsm/L(对于190NaCl)。细胞内溶液含有(以mM为单位)：120葡萄糖酸钾、20TEA-Cl、2MgCl2、2Na2ATP、1EGTA和10HEPES，用KOH调节至pH 7.3(～281mOsm/L)。
在全细胞膜片钳实验前一小时，将表达NaX或NaX-QTT-eGFP-2×FLAG的HEK293T细胞接种在玻璃盖玻片上。对于更接近生理的条件(图4D和7B)，细胞外溶液含有150mM NaCl、5mM KCl、0.5mM CaCl2、1.2mM MgCl2、10mM HEPES和13mM D-(+)-葡萄糖，用NaOH调节至pH7.4，～320mOsm/L；且细胞内溶液含有140mM CsCl、10mM CsF、5mM EGTA、10mM HEPES和2mMNa2ATP，用CsOH调节至pH 7.2，～304mOsm/L。为了确定NaX-QTT是否具有阳离子渗透性，我们首先用NMDG+代替所有细胞外阳离子(图4E)。细胞外溶液含有150mM NMDG和10mM HEPES(相应地添加D-(+)-葡萄糖以达到渗透压～325mOsm/L，并用盐酸将pH调节至7.4)。为了确定NaX-QTT是否具有阴离子渗透性，我们用大阴离子甲磺酸盐(MS-)取代细胞内和细胞外溶液中的所有Cl-离子(图15C)。细胞外溶液含有150mM NaMS和10mM HEPES(相应地添加D-(+)-葡萄糖以达到渗透压～325mOsm/L，并用NaOH将pH调节至7.4)。细胞内溶液含有136mMNaMS、10mM NaF、5mM EGTA、10mM HEPES和2mM Na2ATP(用NaOH调节至pH 7.2，～309mOsm/L)。细胞外NMDG溶液含有150NMDG和10mM HEPES(相应地添加D-(+)-葡萄糖以达到渗透压～325mOsm/L，并用甲磺酸将pH调节至7.4)。
对于离子选择性实验(图4E和5A)，(i)涉及单价阳离子，细胞外溶液含有150mMXCl、10mM HEPES，并相应地添加D-(+)-葡萄糖以达到渗透压～325mOsm/L，并使用XOH将pH调节至7.4，其中X指示感兴趣的阳离子(Na+、K+、Li+或Cs+)；(ii)涉及Ca2+，细胞外溶液含有110mM CaCl2、10mM HEPES，并相应地添加D-(+)-葡萄糖以达到渗透压～325mOsm/L，并使用Ca(OH)2将pH调节至7.4；(iii)细胞内溶液含有136mM NaCl、10mM NaF、5mM EGTA、10 mMHEPES和2mM Na2ATP，用NaOH调节至pH 7.2，～309mOsm/L。
实例1.12-数据分析
对于数据分析，原始电流轨迹通常使用8极贝塞尔(Bessel)低通滤波器在500-800Hz进行过滤。电流轨迹经历5倍数据缩减以显示在图中。数据表示为来自至少两批细胞的至少3个细胞的平均值±标准偏差(SD)。对于使用Na+细胞内溶液进行的离子选择性实验(图4E和5A)，参考Na+细胞内溶液测量液体接界电位(LJP)，并在记录后进行校正。相对离子渗透率(PNa/PX)使用方程式PX/PNa＝[Na+]IC exp(ErevF/RT)/[X+]EC(X＝Na、K、Li或Cs)计算，其中F是法拉第常数，R是气体常数，T＝274K，且Erev从-80-mV到+80-mV斜坡测量(针对LJP进行校正)。
实例2-人Nax功能的评估
在响应于膜电压的变化而在HEK293T细胞或非洲爪蟾卵母细胞中表达人Nax后，未检测到可靠的离子电流(图1A-1D、7A-7D和8A-8D)。在卵母细胞的不同电压阶跃程序中，经典NaV通道活化剂如乌头碱、藜芦定碱、蛛蜂毒素(pompilidotoxin)、降血压物质-I和神经毒素-II的施加并未引起任何明确的人NaX介导的电流(图1B、1D和8A-8B)。尽管人NaX的表面表达强劲，但与NaV或电压门控钙(CaV)通道辅助亚基(即β1、β3、α2δ1、γ2)的共表达也未能产生可靠的电流(图1C、1D、7A、7C和8C-8D)。这些结果大大扩展了先前的发现，即NaX不能作为传统的电压门控通道进行操作14，15。
据报道，细胞外钠浓度[Na+]e的增加会活化表达鼠NaX的神经胶质细胞和神经母细胞瘤(Neuro-2a)细胞系中的非失活Na+电流。还表明鼠NaX与质膜上的Na+/K+-ATP酶和突触相关蛋白97(SAP97)相互作用。尽管具有表面表达且我们具备也将ATP1A1α-亚基在一些条件(表2)下共免疫沉淀的能力，但在曝露于200mM[Na+]e时，在共表达人NaX与Na+/K+-ATP酶亚基和SAP97的卵母细胞中未检测到可靠的内向Na+电流(图1C和1D)。
表2：所分析的Nax通道样品
SCN7A+SCN1B
SCN7A+SCN3B
当人NaX在Neuro-2a细胞中表达时，我们观察到在全细胞膜片钳配置中[Na+]e从140mM变化到190mM时会出现小而闪烁的内向电流(图1E-1G和7E-7F)。表达NaV1.7通道的Neuro-2a细胞在渗透压变化后表现出类似的闪烁行为，表明这种现象并非表达人NaX的细胞所特有(图1E-1G)。仅当密封电阻低(＜1GΩ)时才观察到非失活内向电流，并且在模拟转染和经人NaX转染的Neuro-2a细胞中均发现了这种表型(图7F)。总体而言，尚不清楚我们的研究结果是否反映了人与小鼠NaX通道之间的功能趋异性(图9)，或实验程序的细微差异，但我们未能通过人NaX测得任何可靠电流。我们接下来考虑了对人NaX的结构确定是否可以提供对其功能或调节的洞察。
实例3-脂质纳米盘中β3-Nax复合物的结构确定
人NaX在HEK293细胞中重组地表达，并在温和的洗涤剂葡糖-薯蓣皂苷元中纯化(图10A-10C)。受Nav1.7通道与辅助β-亚基形成稳定的复合物的报道的启发23-25，将β1或β3亚基共表达并发现它们与NaX共纯化(表2)。为了提供更自然的环境，β3-NaX复合物使用常见的磷脂混合物重建在纳米盘中(图10A-10C)。在样品玻璃化之前，将Na+浓度提高到报道的活化NaX的阈值以上，达到200mM。由此产生的人β3-Nax复合物的cryo-EM重建扩展至分辨率(图11A-11F、12A和表3)，并代表了在重建脂质膜环境中确定的真核NaV通道家族成员的第一结构。首先，下列表3的标题行，β3-NaX-纳米盘的EMDB参考是EMDB-25919，且PDB参考是PDB 7TJ8。β3-Nax-GDN的EMDB参考是EDMB-25920且PDB参考是PDB 7TJ9。
表3：cryo-EM数据收集、细化和验证统计
实例4-β3-Nax通道复合物的整体结构
β3-NaX通道复合物类似于四叶草，其中VSLD排列在中央孔模块周围的结构域调换组织中(图1H)，其符合NaV、CaV和NALCN通道之间共享的架构。在Nax中，可溶性细胞内胺基末端结构域停靠在VSLD1下方，细胞内DIII-DIV接头结合在孔旁边，而细胞内羧基末端结构域(CTD)没有得到很好的解析(图13A)，特征让人联想到可用的人NaV通道结构。在细胞外侧，发现两个和四个N连接聚糖分别从NaX和β3亚基延伸。
β3-亚基通过在NaX孔模块的精致型细胞外环之间楔入其免疫球蛋白结构域并将其单个跨膜螺旋定位于VSLD3，形成广泛的相互作用(图1H和13B)。尽管当前的β1或β3辅助亚基似乎与NaX形成稳定的复合物(表2)，但其孔模块不具有与β2或β4亚基形成共价复合物所需的半胱氨酸残基23，27。
在脂质纳米盘中重组的人NaX的整体结构与在洗涤剂中确定的NaV1.1、NaV1.2、NaV1.4、NaV1.5和NaV1.7通道结构高度相似23，28-31(图13C)。相反，NaX确实在经常与NaV通道门控有关的区域内显示出明显的偏差，包括VSLD4的位置偏移，以及沿DII S4-S5连接器的位移(图13D和13E)。观察到的NaX结构变化似乎是其高序列趋异性的结果，并有助于其独特的功能(图9)。
实例5-NaX孔模块处于非导电状态
NaX孔模块含有外前庭、离子选择性过滤器、中央腔和S6活化门(图2A)。S6门很窄并由疏水侧链的双环衬砌，而疏水侧链将会形成水合离子通过的屏障，这明确定义了非导电状态(图2A、2B和13C)23,28-31。然而，由于我们的NaX结构是在高Na+浓度下确定的，因此这些发现似乎与鼠NaX形成Na+活化的非失活通道的报道不一致。相反，观察到的孔模块的非导电状态与我们对人NaX的生理研究的预测完全一致(图1A-1F、7A-7F和8A-8D)。
NaX中央腔的体积与NaV通道中的那些相似，但孔阻断药物通常所靶向的DIV-S6苯丙氨酸32，33经DIV-Trp1484置换(图13F)。NaX揭示了四个穿透孔模块的横向开窗，鉴定了疏水性药物或脂质直接进入中央腔的膜进出通路(图2C和2D)，如在NaV通道中首次提出28-30,34。因此可以想象，经典的NaV或CaV通道拮抗剂可能靶向中央腔内的Nax。
实例6-Nax DIII-DIV接头限制S6门扩张
Nax的细胞内DIII-DIV接头以非常类似于人NaV通道的快速失活状态结构的方式与孔模块受体位点相互作用(图2B和14A)。NaX IFI基序结合在DIII-S4-S5接头与DIV-S5和S6之间，类似于限制NaV通道中S6门扩张的复合物(图2B和14A)。尽管长期以来认为NaX的DIII-DIV接头是一个具有实质性序列趋异性的位点(图14B)，但它显然与NaV通道中的这个关键功能区具有高度的结构同源性(图14A)。尽管如此，将NaX的IFI基序突变为经典的非失活NaV通道QQQ基序不足以在经注射的卵母细胞中产生功能型通道(图14C)，这进一步突出了对NaX通道进行门控的不同要求的存在。
我们利用NaX和NaV1.7结构模型构建了一组精确靶向的嵌合通道，以评估导致人NaX非导电状态的决定因素。双结构域DII-DIII和DIII-DIV NaV1.7-NaX嵌合体和单结构域DIII NaV1.7-NaX嵌合体在爪蟾卵母细胞中单独表达时产生大量向内和向外电流(图2E)。最终，调换NaV1.7-VSD3和相关的DIII-S4-S5接头两者是足以转移泄漏通道表型的最小区域(图14D)。因为这些区域是形成IFM基序的受体位点(其限制NaV通道中的S6扩张)所需的(图14A)，并且它们取代到NaX中会产生强劲的泄漏电流(图2E和14D)，我们建议DIII-NaX的DIV接头用于限制S6扩张并稳定孔模块的非导电状态。
实例7-NaX孔经膜脂质漫润
根据定义，离子通道的关闭或失活状态是不导电的。在β3-NaX结构中，四个脂质穿过横向开窗进入狭窄的疏水S6门上方的中央腔。这些惊人的密度可以明确指定为三个磷脂和一个胆固醇(图2C和2D)。一个磷脂甚至横跨并密封细胞内S6活化门(图2C和14E)。
为了评估脂质纳米盘环境是否导致β3-Nax中膜脂质的强劲可视化，我们在没有外源性磷脂补充的洗涤剂葡糖-薯蓣皂苷元中制备了一个新样品，并将其结构确定为分辨率(图11G-11L、12B和表3)。我们观察到相同的NaX孔结构被四个明确定义的共纯化脂质穿透(图14F)。因此，β3-NaX中的离子传导通路在该孔模块构形中是明确不导电的，因为它被脂质渗透并封闭(图2A-2D和14E-14F)。结合在Nax内的解析良好的共纯化脂质的存在可能反映了其非导电状态的内在稳定性以及S6门和中央腔的疏水性。
实例8-所靶向的孔润湿突变可以活化人Nax
基于我们的总体观察结果，即S6门是狭窄的、疏水的和脂质结合的，并且IFI基序受体位点的破坏可能促进S6扩张，我们推断在S6门周围有针对性地引入极性残基可能促进孔水合、结合脂质的不稳定化以及NaX转变为开放的导电状态(图3A-3F)。据此，当响应于电压阶跃程序而在非洲爪蟾卵母细胞中单独表达时，在三个S6门衬砌位置处含有极性取代的三突变型NaX通道构建体(F724Q-I1189T-I1492T)，即NaX-QTT，产生强劲的离子电流(图3A和3B)。对应的单点突变型NaX通道构建体未能产生强劲的电流，表明存在一个孔润湿阈值，以通过非导电孔模块状态的不稳定来实现S6门扩张(图3C)。L390E-I189E-I1492E三突变型NaX通道构建体(NaX-EEE)也显示出功能，但相对于NaX-QTT，具有更低的电流振幅和更高的电流可变性(图3D和3F)。
在经注射的卵母细胞中，NaX-QTT电流外向整流，即使在长(5秒)去极化测试脉冲期间也没有失活迹象(图3B)。因为NaX可以与β1或β3亚基稳定地相互作用(图1H和表2)，所以评估了这些辅助亚基的功能效应。当NaX-QTT分别在p1或β3亚基的存在下表达时，没有观察到电流振幅或动力学的差异(图3E和3F)。在HEK293T细胞中，当与C末端GFP-2xFLAG标签(见下文)一起表达时，NaX-QTT在全细胞膜片钳实验中也表现为非失活通道。至关重要的是，Nax-QTT通道构建体仅在细胞内S6门周围含有三个被靶向的突变(图3A)，远离选择性过滤器和其他重要通道区，因此下面我们将Nax-QTT表征为代表以评估人NaX通道的离子选择性和药理学。
实例9-NaX选择性过滤器
独特的Nax的离子选择性过滤器(DENA基序)由局部架构支持，该架构与支持NaV通道中的经典DEKA基序的支架存在根本性差异(图4A-4C)。NaV通道中的四个标志性色氨酸残基形成涉及三个保守苏氨酸侧链的结构域间相互作用(图4B)23，28-30，35，36。在NaX中，Pro355置换DI色氨酸，而Ala351和Ile1432分别取代DI和DIV苏氨酸对应物(图4B)。在NaV1.5中，在等效于NaX Pro355和Ile1432的位置处的取代是致病性的，突出了这些改变的潜在功能相关性。因此，NaX选择性过滤器比NaV通道中的选择性过滤器更宽且更具电负性(图4B)。
在NaX的DENA基序处，Asp353(DI)与DII-Trp689的骨架形成氢键，Glu688(DII)指向离子渗透通路内，且Asn1142(DIII)与DIV-Phe1433羰基键合以将的径向移动(相较于NaV通道中的DIV丙氨酸)施加于Ala1434(DIV)上(图4A和4C)。Ala1434的相对偏移由Nax中的DI-Pro355调节，这也允许DI-Tyr359重新定向～90°以填充由不存在的常规DI色氨酸空出的体积(图4C)。这种结构重塑使外前庭中的DI-Glu356和DII-Glu691位移了这改变了典型河豚毒素(TTX)结合位点，并可能突出发现NaX选择性抑制剂的潜力(图15A)。
实例10-Nax不辨别单价阳离子
相对于在建立NaV通道中的Na+选择性中发挥关键作用的传统DIII-赖氨酸，标志性的DIII-Asn1142侧链显著改变了NaX选择性过滤器的结构和化学特征(图4A-4C)。据此，我们通过单个突变将DENA基序引入人NaV1.7通道中，并观察到经注射的爪蟾卵母细胞中Na+和Li+选择性的显著损失，但记录了具有异常动力学的可测量的K+电流(图15B)。在交互实验(reciprocal experiment)中，典型DEKA基序到NaX的突变转移没有产生可测量的电流(图15B)，证实了NaX DENA基序之外的分子特征有助于其活化。
我们检查了来自在HEK293T细胞中表达的NaX-QTT构建体的电流，并且发现当细胞外溶液中的所有阳离子均由大阳离子N-甲基-D-葡糖胺(NMDG)置换时，反转电位从+5mV位移到低于-100mV(图4D)，证实该构建体表现为阳离子通道，并且大阳离子不渗透。用大阴离子甲磺酸根置换所有细胞外Cl-离子未改变电流特征(图15C)，表明NaX表现为阳离子选择性通道，这与选择性过滤器的尺寸和电负性特征相一致(图4B)。正如预期的那样，使用斜坡程序(-80至+80mV)和简单Na+细胞内溶液(150mM)，在对称NaCl/NaCl条件下，反转电位接近0mV(图4E)。值得注意的是，当细胞暴露于不同的细胞外单价阳离子(150mM)时，仅测量到反转电位的轻微变化，证实NaX-QTT不能有效地辨别Na+、K+、Cs+或Li+离子(图4E)
实例11-通过细胞外Ca2+抑制Nax
我们测量了来自NaX-QTT的电流并观察到Ca2+并未渗透(图5A)。然而，细胞外Ca2+显示出抑制效应，其中生理相关浓度的Ca2+(1mM)抑制了～70％的外向Na+电流(图5A)。值得注意的是，相对于传统的NaV通道，细胞外Ca2+阻断对NaX的效力高～10倍，证实了选择性过滤器内的物理化学差异(图4A和4B)。二价阳离子Zn2+和Co2+在1mM时抑制NaX-QTT电流，而三价阳离子Gd3+的抑制作用更强(图5B)，这表明由多价阳离子造成的孔阻断基本上不受更宽且电负性更高的Nax选择性过滤器结构的影响(图4B)。
实例12-Nax对经典Nav通道阻断剂敏感
我们测试了一系列经典的NaV通道阻断剂，以确定NaX-QTT的药理特性。值得注意并且与之前关于NaX是TTX抗性通道的报道形成鲜明对比的是，河豚毒素在低至10nM的浓度下显示出对NaX-QTT的抑制效应(图5C)。TTX阻断的效力表明它发生在选择性过滤器处，与敏感性NaV通道亚型的TTX阻断中所涉及的侧链的实质性保守相一致(图9和15A)。
全身麻醉剂利多卡因在低mM浓度下有效阻断NaX-QTT电流(图5C)，可能是通过在通道的中央腔内结合。奎尼丁和洛哌丁胺(loperamide)等其他经典的孔阻断药物在高μM浓度下同样有效地抑制NaX-QTT电流(图5C)。相比之下，氟卡尼、雷诺嗪(ranolazine)或苯妥英(phenytoin)在浓度高达300μM时未观察到对NaX-QTT电流的强烈抑制作用(图5C)。
实例13-Nax揭示了非典型的电压传感器样结构域
尚未确定鼠或人NaX通道在何种条件下产生电压活化电流，这种电压敏感性的明显缺乏仍然是一个谜。矛盾的是，人NaX中S4门控电荷的数量仅适度减少14，15，22(图9)，且其VSLD与Nav和CaV通道的VSD具有高度的整体结构相似性(图13D和13E)。有趣的是，虽然NaX中的VSLD1、VSLD2和VSLD3都存在于活化的构形中，但VSLD4是失活的(图6A-6D)。对NaX VSLD进行更深入的评估会提供对其明显的电压敏感性缺陷的洞察。
相对于人NaV通道，NaX中的VSLD1仅含有四个预期S4门控电荷中的三者(203PTLQTARTLRILKIIP218；SEQ ID NO：87；图6A和9)。K4由疏水构建位点(HCS)下方的细胞内负电荷簇(INC)协调，类似于活化的构形(图6A)37，38。值得注意的是，已知在R1处的谷氨酰胺中和会降低其他通道中的电压敏感性40，并且在Nav通道中作为丝氨酸保守的位置处，Pro203在细胞外S3-S4环内产生径向弯曲，这可能影响NaX中的S4运动(图6A和9)。
NaX VSLD2含有所有五个预想的S4门控电荷(593ALLRLFRMLRIFKLGK 608；SEQ ID NO：88)，并且其活化状态似乎由与S4羰基骨架键合的独特的S3 His579侧链稳定化(图6B和9)。此外，相对于人Nav通道结构，DII S4-S5接头向胞质溶胶的径向位移可能改变VSLD2偶合(图6B和13D)。这是通过NaX DII-S6螺旋中的单个残基缺失传播的，它重新定位Phe731以进行～180°重新定向以强制S4-S5接头偏移(图6B和9)。
NaX VSLD3仅显示六个典型S4门控电荷(1024 KPLISMKFLRPLRVLSQ 1040；SEQ ID NO：89)中的四者，这种改变预计会稳定其活化的构形并降低内在的电压敏感性(图6C和9)。此外，Phe1011(S3)显著且独特地扩展了VSLD3中的HCS(图6C和9)，预计这将进一步影响S4门控电荷运动。
NaX VSLD4仅含有四个S4门控电荷(13466 QLILLSRIIHMLRLGKGPKVFH 1367；SEQ ID NO：90)，因为H4和H8在生理pH下不可能带电37,39,44,45(图6D和9)。相比之下，典型NaV通道中的VSD4和相关的DIV S4-S5接头保留了八个与快速失活机制密不可分的S4门控电荷。与人NaV通道结构23,28,29,31，37和在0mV的预期形成鲜明对比的是，Nax的VSLD4以类似失活的状态存在，如通过S4向胞质溶胶的相对偏移所示(图6D和16A)。这一观察结果令人惊讶，但与迄今为止针对NaX描述的电压敏感性缺陷相一致。
实例14-探究趋异型Nax VSLD的功能
在NaX的VSLD中鉴定的结构特征表明对通道功能的非典型贡献。此外，许多突出的序列特性发生在NaX的据报道为相关人通道中的致病改变的位置处。NaX VSLD11中的Pro203等效于NaV1.7中的致病性Ser211Pro突变，其据报道可使通道活化超极化-12mV(图6A和16B)。基于结构的序列比对表明，类似的框内DII-S6缺失在NaV1.7(ΔLeu966)中是致病性的，并产生-16mV的活化超极化(图6B和16B)。NaX VSLD3中的Phe1011对应于KCNQ1中的致病性Ser209Phe突变，其据报道可对通道活化产生强烈的超极化(-45mV)(图16C)43，而NaV1.7中的等效Ser1269Phe取代产生了～8mV的去极化位移(图6C和16B)。此外，三个脯氨酸独特地存在于NaX的VSLD4中(图9和16D)，并且NaV1.5中的对应VSD4残基突变为脯氨酸赋予稳态失活强烈的去极化位移(15mV)，这与在VSLD4中观察到失活的构形相一致(图6D、16A和16E)。
为了进一步探究NaX VSLD功能，我们通过将单个VSLD定向转移到NaV1.7中来设计嵌合构建体。值得注意的是，记录了来自表达NaV1.7-NaX-VSLD2通道嵌合体的经注射的非洲爪蟾卵母细胞的强劲电流(图6E)，指示VSLD2要么是经组成性活化的，要么本质上能够进行机电偶合。相比之下，NaX-VSLD1、VSLD3或VSLD4的批量转移使Nav1.7嵌合通道失去功能(图6E)。因此，尽管NaX与NaV、Cav和KV通道共享全局结构相似性，但我们的研究结果表明，累积序列适应可能导致跨通道超家族的高度趋异的功能结果。
实例15-示例性Nax/Nav1.7嵌合构建体
如下表4中所示，构建了一系列嵌合型人NaX蛋白，其中插入了来自人NaV1.7的一个或多个区域以代替对应的NaX序列。如果表达构建体的非洲爪蟾卵母细胞在从0mV的保持电位开始在+80mV或-100mV引发的电流振幅显著高于来自表达人NaX野生型的卵母细胞的电流振幅(参见图18)，则认为该构建体是功能性的(参见表4)。关于表4的前7个嵌合体和hNaX-hNaV1.7DIV嵌合体的数据也在实例6中描述，并显示在图2E中。
讨论
上述实例对结构、功能和药理学进行了综合分析，以提供对NaX通道的深入了解。通过对不同表达系统、辅因子和药理活化剂的综合评估，我们证明了人NaX不能作为电压活化通道发挥作用。导致NaX的电压敏感性改变的分子特征可能包括选择性过滤器、电压传感器和孔模块内的单个残基变化，因为类似的变化会极大地影响NaV1.5和Nav1.7通道的门控特性(图6A-6E、9、16B和16E)。事实上，许多导致NaX结构改变的位置变化似乎是NaV通道中疾病热点位置的基础，但这些观察结果的意义仍然未知。
使用结构引导的蛋白质工程，我们发现整个同源结构域或子结构域(例如VSLD)在NaX与NaV1.7之间的转移会产生非功能性构建体或具有非常不寻常的门控特征的通道(图2E和6E)。这些结果表明，NaX门控的偶合或能量学与NaV通道实质性地趋异，这可能指向这些密切相关的通道支架之间的偶合界面的结构差异。值得注意的是，我们的研究并未排除NaX在特定细胞环境中可能作为电压调节通道的可能性。可以想象，假定的辅助亚基或辅因子的结合可能赋予NaX一定程度的电压敏感性，如最近对于NALCN或TMEM16A通道的描述。
调和Na+感测的差异
据报道，当增加细胞外Na+浓度>150mM时，鼠NaX产生非失活的内向电流。我们仅观察到具有低密封阻力的定性上相似的记录，并且未能在人NaX的异源表达时重建Na+活化电流，即使在与预期将其稳定在质膜上的因子(即SAP97)共同表达时(图1D和7F)。鼠与人NaX蛋白之间的序列趋异性或细胞环境差异可能是这些看似矛盾的功能结果的原因(图9)。
我们的β3-NaX的cryo-EM样品在200mM NaCl中，在高于所报道的Na+依赖性门控所需的阈值9-11的情况下制备。在结构分析时没有观察到明确的Na+感测基因座或基于活化的机制，并且我们可能指定为细胞外Na+离子的唯一图谱特征是在NaX选择性过滤器附近与NaV通道中也发现的位点结合(图17A)29,49。我们推断，针对NaX描述的Na+感测机制很可能是由外源性蛋白因子或通路，诸如Na+/K+ATP酶、内皮素受体或尚未鉴定的系统促成的。可以想象，沿NaX DIII-DIV接头(图14B)的预计磷酸化位点的转译后修饰可能有助于将IFI基序从其孔模块受体位点移开，以促进S6扩张和通道门控。或者，在Nax的DIV和VSLD4中发现的不常见但高度保守的组氨酸残基的存在也可能引发对潜在细胞内二价阳离子敏感或pH敏感的活化或调节机制的推测(图6D和9)。
NaX假设：非选择性Na+泄漏通道
在结构分析和反复性蛋白质工程实验的指导下，我们发现在疏水性S6门区周围的三个残基的靶向突变足以通过在非洲爪蟾卵母细胞和HEK293T细胞中表达的人NaX产生强劲的离子电流(图3、4D、4E和5)。据推测，在NaX-QTT通道构建体中引入三个极性侧链将使S6门水合，去除结合的膜脂质，并允许孔扩张和离子传导(图2B-2D)。这一概念和技术上的突破使我们首次能够表征人NaX的基本特性。
与NaV通道不同，NaX-QTT在细胞外Ca2+离子不存在下作为欧姆泄漏通道有效地传导单价阳离子(图3、4D、4E)。Ca2+不渗透但阻断NaX-QTT，因此在生理含量的细胞外Ca2+存在下，NaX-QTT电流外向整流，在负膜电位下仅有少量内向泄漏组分(图5A)。这种独特的电流表型具有针对NaV样通道预期的药理学敏感性，包括通过TTX、Zn2+、Co 2+和Gd3+的明显选择性过滤器阻断，以及通过利多卡因、奎尼丁和洛哌丁胺的假定孔阻断(图5B和5C)。氟卡尼、雷诺嗪或苯妥英没有对NaX-QTT电流产生强烈的抑制作用(图5C)，表明人NaX相对于典型NaV通道具有独特的药理学特征，并且可能反映了独特的中央腔衬砌残基如DIV-Trp1484(图13F)，或Nax中可能不存在失活状态(图3B和4D)。尽管Nax-QTT电流是非失活的并且不受电压调节，但未来的研究将需要在野生型人Nax通道中定义这些重要特征，因为孔润湿QTT突变被靶向至S6门区。尽管如此，值得注意的是，早期的一项研究表明Nax在体内确实起到了泄漏通道的作用。
NaX选择性过滤器结构的物理、化学和静电特征符合阳离子选择性通道的预期(图4A和4B)，但它缺乏在NaV通道中赋予Na+选择性的DIII-赖氨酸侧链(图9)。事实上，我们对NaX-QTT的电生理学表征显示，在所检查的单价阳离子(Na+、K+、Cs+和Li+)中没有选择性(图4E)。因此，我们假设人NaX作为Na+泄漏通道发挥作用，该通道在生理条件下对细胞外Ca2+的调节敏感(图5A)。值得注意的是，NaX-QTT的Ca2+阻断比NaV通道上的Ca2+阻断强～10倍(图5A)，但类似于略微相关的非选择性NALCN Na+泄漏通道的Ca2+阻断敏感性，这可能反映了一个共享的控制机制以避免过度去极化Na+流入细胞。
可视化孔结合的膜脂质
我们在洗涤剂或脂质纳米盘中确定的NaX通道结构的一个惊人特征是非导电状态的可视化，其中脂质完全阻塞离子传导通路并密封S6门(图2C、2D和14F)。堵塞S6门的磷脂让人联想到在NaV通道结构中结合在S6门上的胆固醇样分子28，29，31(图17B)。这些疏水性配体在S6门处的占据似乎与保守的DIV-S6酪氨酸侧链的“向上”构形相关，该构形传统上涉及NaV通道中的药物阻断(图9和17B)23，31-33。值得注意的是，中央腔内的药物结合可以强制这种保守的S6酪氨酸的“向下”构形30，31，同时在S6门处发生类脂密度的偏移(图17B)31。因此，我们的NaX结构支持一种新兴范式，通过该范式，膜脂可以密封S6门，并且脂质占据可以调节离子电导、药物结合和孔结构。这种发展中的模型与长期以来的S6门必须在NaV通道中紧密关闭并完全关闭30的观点形成鲜明对比。
总结与展望
NaX通道的基本生理学和药理学特性自其在近30年前被克隆以来一直是个谜。我们发现，在我们所研究的条件下，人NaX既不能作为Na+活化通道独立地发挥作用，也不能作为电压活化通道独立地发挥作用。通过结构引导的蛋白质工程工作，我们发现了一种使用传统异源表达系统来表征人NaX通道变体的方法。NaX-QTT通道构建体显示出独特但类似于NaV通道的药理学特征(包括对Ca2+和TTX的有效抑制)，并显露出非选择性阳离子电导，表明NaX通道可能在体内作为Ca2+调节的Na+泄漏通道起作用。总体而言，这项研究为进一步探究NaX的生理学和药理学提供了必要的工具，并极大地促进了我们对这种非典型离子通道的理解。
参考文献
1Noda，M.&Hiyama，T.Y.Sodium sensing in the brain.Pflugers Arch 467，465-474，doi：10.1007/s00424-014-1662-4(2015)。
2Andersson，B.Thirst--and brain control of water balance.Am Sci 59，408-415(1971)。
3Liamis，G.等人Electrolyte disorders in community subjects：prevalenceand risk factors.AmJMed 126，256-263，doi：10.1016/j.amjmed.2012.06.037(2013)。
4Adrogue，H.J.Consequences of inadequate management of hyponatremia.AmJ Nephrol25，240-249，doi：10.1159/000086019(2005)。
5Watanabe，E.等人Nav2/NaG channel is involved in control of salt-intake behaViorin the CNS.J Neurosci 20，7743-7751(2000)。
6Nomura，K.等人[Na(+)]Increases in Body Fluids Sensed by Central NaXInduce Sympathetically Mediated Blood Pressure EleVations Via H(+)-DependentActiVation of ASIC 1a.Neuron 101，60-75e66，doi：10.1016/j.neuron.2018.11.017(2019)。
7Hiyama，T.Y.等人Autoimmunity to the sodium-level sensor in the braincauses essential hypernatremia.Neuron66，508-522，doi：10.1016/j.neuron.2010.04.017(2010)。
8Xu，W.等人Sodium channel NaX is a regulator in epithelial sodiumhomeostasis.Sci Transl Med 7，312ra177，doi：10.1126/scitranslmed.aad0286(2015)。
9Hiyama，T.Y.等人Na(x)channel involved in CNS sodium-level sensing.NatNeurosci 5，511-512，doi：10.1038/nn0602-856(2002)。
10Shimizu，H.等人Glial NaX channels control lactate signaling toneurons for brain[Na+]sensing.Neuron54，59-72，doi：10.1016/j.neuron.2007.03.014(2007)。
11Hiyama，T.Y.等人Endothelin-3expression in the subfornical organenhances the sensitiVity of Na(x)，the brain sodium-leVel sensor，to suppresssalt intake.Cell Metab 17，507-519，doi：10.1016/j.cmet.2013.02.018(2013)。
12Matsumoto，M.等人Channel properties of NaX expressed in neurons.PLoSOne 10，e0126109，doi：10.1371/journal.pone.0126109(2015)。
13Noda，M.&Hiyama，T.Y.The Na(x)Channel：What It Is and What ItDoes.Neuroscientist21，399-412，doi：10.1177/1073858414541009(2015)。
14Akopian，A.N.，SouSlova，V.，Sivilotti，L.&Wood，J.N.Structure anddistribution of a broadly expressed atypical sodium channel.FEBSLett400，183-187，doi：10.1016/s0014-5793(96)01389-0(1997)。
15Felipe，A.，Knittle，T.J.，Doyle，K.L.&Tamkun，M.M.Primary structure anddifferential expression during development and pregnancy of a novel voltage-gated sodium channelin the mouse.J Biol Chem 269，30125-30131(1994)。
16Goldin，A.L.等人Nomenclature of voltage-gated sodium channels.Neuron28，365-368，doi：10.1016/s0896-6273(00)00116-1(2000)。
17Noda，M.等人Existence of distinct sodium channel messenger RNAs inrat brain.Nature 320，188-192，doi：10.1038/320188a0(1986)。
18Shen，H.等人Structure of a eukaryotic voltage-gated sodium channelat near-atomic resolution.Science 355，doi：10.1126/science.aal4326(2017)。
19Chanda，B.&Bezanilla，F.Tracking voltage-dependent conformationalchanges in skeletal muscle sodium channel during activation.J Gen Physiol120，629-645，doi：10.1085/jgp.20028679(2002)。
20Capes，D.L.，Goldschen-Ohm，M.P.，Arcisio-Miranda，M.，Bezanilla，F.&Chanda，B.Domain IV voltage-sensor movement is both sufficient and ratelimiting for fast inactivation in sodium channels.J Gen Physiol 142，101-112，doi：10.1085/jgp.201310998(2013)。
21Clairfeuille，T.等人Structural basis of alpha-scorpion toxin actionon NaV channels.Science 363，doi：10.1126/science.aav8573(2019)。
22George，A.L.，Jr.，Knittle，T.J.&Tamkun，M.M.Molecular cloning of anatypical voltage-gated sodium channel expressed in human heart and uterus：evidence for a distinct gene family.Proc Natl Acad Sci U S A89，4893-4897，doi：10.1073/pnas.89.11.4893(1992)。
23Shen，H.，Liu，D.，Wu，K.，Lei，J.&Yan，N.Structures of human Nav1.7channelin complex with auxiliary subunits and animal toxins.Science 363，1303-1308，doi：10.1126/science.aaw2493(2019)。
24Laedermann，C.J.，Syam，N.，Pertin，M.，Decosterd，I.&Abriel，H.beta1-andbeta3-voltage-gated sodium channel subunits modulate cell surface expressionand glycosylation of Nav1.7in HEK293 cells.Front Cell NeuFosci 7，137，doi：10.3389/fncel.2013.00137(2013)。
25Kanellopoulos，A.H.等人Mapping protein interactions of sodiumchannel NaV1.7using epitope-tagged gene-targeted mice.EMBO J 37，427-445，doi：10.15252/embj.201796692(2018)。
26Sokolov，M.V.等人Co-expression of beta Subunits with the Voltage-Gated Sodium Channel NaV 1.7：the Importance of Subunit Association andPhosphorylation and Their Effects on Channel Pharmacology and Biophysics.JMol Neurosci 65，154-166，doi：10.1007/s12031-018-1082-6(2018)。
27Das，S.，Gilchrist，J.，Bosmans，F.&Van Petegem，F.Binary architecture ofthe Nav1.2-beta2 signaling complex.Elife 5，doi：10.7554/eLife.10960(2016)。
28Pan，X.等人Structure of the human voltage-gated sodium channelNav1.4in complex with betal.Science 362，doi：10.1126/science.aau2486(2018)。
29Pan，X.等人Molecular basis for pore blockade of human Na(+)channelNav1.2by the mu-conotoxin KIIIA.Science 363，1309-1313，doi：10.1126/science.aaw2999(2019)。
30Jiang，D.等人SStructure of the Cardiac Sodium Channel.Cell180，122-134e110，doi：10.1016/j.cell.2019.11.041(2020)。
31Li，Z.等人Structural basis for pore blockade of the human cardiacsodium channel Nav1.5by tetrodotoxin and quinidine.bioRxiv doi.org/10.1101/2019.12.30.890681(2020)。
32Ragsdale，D.S.，McPhee，J.C.，Scheuer，T.&Catterall，W.A.Moleculardeterminants of state-dependent block of Na+channels by localanesthetics.Science 265，1724-1728，doi：10.1126/science.8085162(1994)。
33Pless，S.A.，Galpin，J.D.，Frankel，A.&Ahern，C.A.Molecular basis forclass Ib anti-arrhythmic inhibition of cardiac sodium channels.Nat Commun 2，351，doi：10.1038/ncomms1351(2011)。
34Hille，B.Local anesthetics：hydrophilic and hydrophobic pathways forthe drug-receptor reaction.JGen Physiol69，497-515，doi：10.1085/jgp.69.4.497(1977)。
35Payandeh，J.，Scheuer,T.，Zheng，N.&Catterall，W.A.The crystal structureof a voltage-gated sodium channel.Nature 475，353-358，doi：10.1038/nature10238(2011)。
36Payandeh，J.，Gamal El-Din，T.M.，Scheuer，T.，Zheng，N.&Catterall，W.A.Crystal structure of a voltage-gated sodium channel in two potentiallyinactiVated states.Nature 486，135-139，doi：10.1038/nature11077(2012)。
37Ahuja，S.等人Structural basis of Nav1.7inhibition by an isoform-selective small-molecule antagonist.Science 350，aac5464，doi：10.1126/science.aac5464(2015)。
38Tao，X.，Lee，A.，Limapichat，W.，Dougherty，D.A.&MacKinnon，R.A gatingcharge transfer center in voltage sensors.Science 328，67-73，doi：10.1126/science.1185954(2010)。
39Gagnon，D.G.&Bezanilla，F.A single charged voltage sensor is capableof gating the Shaker K+channel.J Gen Physiol 133，467-483，doi：10.1085/jgp.200810082(2009)。
40Kontis，K.J.，Rounaghi，A.&Goldin，A.L.Sodium channel activation gatingis affected by substitutions of voltage sensor positiVe charges in all fourdomains.J Gen Physiol 110，391-401，doi：10.1085/jgp.110.4.391(1997)。
41Estacion，M.等人Can robots patch-clamp as well as humans？Characterization of a novel sodium channel mutation.J Physiol 588，1915-1927，doi：10.1113/jphysiol.2009.186114(2010)。
42Yang，Y.，Estacion，M.，Dib-Hajj，S.D.&Waxman，S.G.Molecular architectureof a sodium channel S6 helix：radial tuning of the voltage-gated sodiumchannel 1.7activation gate.J Biol Chem 288，13741-13747，doi：10.1074/jbc.M113.462366(2013)。
43Eldstrom，J.等人Mechanistic basis for LQT1 caused by S3 mutations inthe KCNQ1 subunit of IKs.J Gen Physiol 135，433-448，doi：10.1085/jgp.200910351(2010)。
44Kuhn，F.J.&Greeff,N.G.Movement of voltage sensor S4 in domain 4istightly coupled to sodium channel fast inactivation and gating chargeimmobilization.JGen Physiol 114，167-183，doi：10.1085/jgp.114.2.167(1999)。
45Amold，W.D.等人Defective fast inactivation recovery of NaV 1.4incongenital myasthenic syndrome.Ann Neurol 77，840-850，doi：10.1002/ana.24389(2015)。
46Bosmans，F.，Martin-Eauclaire，M.F.&Swartz，K.J.Deconstructing Voltagesensor function and pharmacology in sodium channels.Nature 456，202-208，doi：10.1038/nature07473(2008)。
47Bosmans，F.，Puopolo，M.，Martin-Eauclaire，M.F.，Bean，B.P.&Swartz，K.J.Functional properties and toxin pharmacology of a dorsal root ganglionsodium channel viewed through its voltage sensors.J Gen Physiol 138，59-72，doi：10.1085/jgp.201110614(2011)。
48Xu，H.等人Structural Basis of Nav1.7Inhibition by a Gating-ModifierSpider ToXin.Cell 176，702-715e714，doi：10.1016/j.cell.2018.12.018(2019)。
49Shen，H.等人Structural basis for the modulation of voltage-gatedsodium channels by animaltoxins.Science 362，doi：10.1126/science.aau2596(2018)。
50Mastronarde，D.N.Automated electron microscope tomography usingrobust prediction of specimen movements.J Struct Biol 152，36-51，doi：10.1016/j.jsb.2005.07.007(2005)。
51Tegunov，D.&Cramer，P.Real-time cryo-electron microscopy datapreprocessing with Warp.Nat Methods 16，1146-1152，doi：10.1038/s41592-019-0580-y(2019)。
52Zivanov，J.等人New tools for automated high-resolution cryo-EMstructure determination in RELION-3.Elife 7，doi：1O.7554/eLife.42166(2018)。
53Grant，T.，Rohou，A.&Grigorieff,N.cisTEM，user-friendly software forsingle-particle image processing.Elife 7，doi：10.7554/eLife.35383(2018)。
54Scheres，S.H.RELION：implementation of a Bayesian approach to cryo-EMstructure determination.J Struct Biol 180，519-530，doi：10.1016/j.jsb.2012.09.006(2012)。
55Zheng，S.Q.等人MotionCor2：anisotropic correction of beam-inducedmotion for improved cryo-electron microscopy.Nat Methods 14，331-332，doi：10.1038/nmeth.4193(2017)。
56Rohou，A.&Grigorieff,N.CTFFIND4：Fast and accurate defocus estimationfrom electron micrographs.J Struct Biol 192，216-221，doi：10.1016/j.jsb.2015.08.008(2015)。
57Matthies，D.等人Single-particle cryo-EM structure of a voltage-activated potassium channel inlipid nanodiscs.Elife 7，doi：10.7554/eLife.37558(2018)。
58Kelley，L.A.，Mezulis，S.，Yates，C.M.，Wass，M.N.&Sternberg，M.J.ThePhyre2 web portal for protein modeling，prediction and analysis.Nat Protoc 10，845-858，doi：10.1038/nprot.2015.053(2015)。
59Emsley，P.，Lohkamp，B.，Scott，W.G.&CoWtan，K.Features and developmentof Coot.Acta Crystallogr D Biol Crystallogr 66，486-501，doi：10.1107/S0907444910007493(2010)。
60Afonine，P.V.等人Real-space refnement in PHENIX for cryo-EM andcrystallography.Acta Crystallogr D Struct Biol 74，531-544，doi：10.1107/S2059798318006551(2018)。
61Goddard，T.D.等人UCSF ChimeraX：Meeting modern challenges inVisualization and analysis.Protein Sci 27，14-25，doi：10.1002/pro.3235(2018)。
62Croll，T.I.ISOLDE：a physically realistic environment for modelbuilding into low-resolution electron-density maps.Acta Crystallogr DStructBiol74，519-530，doi：10.1107/S2059798318002425(2018)。
63The PyMOL Molecular Graphics System，V.r.p.，LLC。
64Pettersen，E.F.等人UCSF Chimera--a visualization system forexploratory research and analysis.JComputChem 25，1605-1612，doi：10.1002/jcc.20084(2004)。
65Chovancova，E.等人CAVER 3.0：a tool for the analysis of transportpathways in dynamic protein structures.PLoS Comput Biol8，e1002708，doi：10.1371/journal.pcbi.1002708(2012)。
66Sievers，F.等人Fast，scalable generation of high-quality proteinmultiple sequence alignments using Clustal Omega.Mol Syst Biol7，539，doi：10.1038/msb.2011.75(2011)。
67Robert，X.&Gouet，P.Deciphering key features in protein structureswith the new ENDscript server.Nucleic Acids Res 42，W320-324，doi：10.1093/nar/gku316(2014)。
68Chua，C.H.，Wulf，M.，Weidling，C.，Rasmussen，L.P.&Pless，S.A.The NALCNchannel complex is voltage sensitiVe and directly modulated by extracellularcalcium.Sci Adv 6，eaaz3154(2020)。
序列表
下表进一步描述本文引用的某些序列。
尽管为了清楚理解起见，通过图示和实例的方式对上述发明进行了详细描述，但是这些描述和实例不应被解释是限制本发明的范围。本文引用的所有专利和科学文献的公开内容均以其全文通过引用明确地并入本文。
序列表
<110> 基因泰克公司
<120> 突变型人 NAX 蛋白和筛选方法
<130> 01164-0016-00PCT
<150> US 63/195,039
<151> 2021-05-30
<160> 90
<170> PatentIn 版本 3.5
<210> 1
<211> 1682
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(1682)
<223> 野生型人 NaX (hNaX) (Uniprot Q01118-1)
<400> 1
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys Thr Cys Cys
915 920 925
Lys Ile Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly Leu Val Thr
930 935 940
Leu Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln
945 950 955 960
Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe
980 985 990
Lys Ala Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val
995 1000 1005
Ile Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg Glu Glu Leu
1010 1015 1020
Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu
1025 1030 1035
Ser Gln Phe Glu Arg Met Lys Val Val Val Arg Ala Leu Ile Lys
1040 1045 1050
Thr Thr Leu Pro Thr Leu Asn Val Phe Leu Val Cys Leu Met Ile
1055 1060 1065
Trp Leu Ile Phe Ser Ile Met Gly Val Asp Leu Phe Ala Gly Arg
1070 1075 1080
Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg Phe Pro Ser
1085 1090 1095
Ser Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu Leu Phe Asn
1100 1105 1110
Glu Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe Asp Asn Val
1115 1120 1125
Gly Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr Phe Asn Gly
1130 1135 1140
Trp Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val Ala Val Asn
1145 1150 1155
Ile Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr Cys Tyr Phe
1160 1165 1170
Ile Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu Ser Met Leu
1175 1180 1185
Ile Thr Val Ile Ile Asp Asn Phe Asn Lys His Lys Ile Lys Leu
1190 1195 1200
Gly Gly Ser Asn Ile Phe Ile Thr Val Lys Gln Arg Lys Gln Tyr
1205 1210 1215
Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser Gln Arg Pro Val
1220 1225 1230
Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe Ile Phe Asp Val Val
1235 1240 1245
Thr Ser Gln Ala Phe Asn Val Ile Val Met Val Leu Ile Cys Phe
1250 1255 1260
Gln Ala Ile Ala Met Met Ile Asp Thr Asp Val Gln Ser Leu Gln
1265 1270 1275
Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile Phe Val Met Leu
1280 1285 1290
Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile Ala Phe Arg Cys Phe
1295 1300 1305
Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe Met Val Val Ile
1310 1315 1320
Phe Ser Ile Thr Gly Leu Cys Leu Pro Met Thr Val Gly Ser Tyr
1325 1330 1335
Leu Val Pro Pro Ser Leu Val Gln Leu Ile Leu Leu Ser Arg Ile
1340 1345 1350
Ile His Met Leu Arg Leu Gly Lys Gly Pro Lys Val Phe His Asn
1355 1360 1365
Leu Met Leu Pro Leu Met Leu Ser Leu Pro Ala Leu Leu Asn Ile
1370 1375 1380
Ile Leu Leu Ile Phe Leu Val Met Phe Ile Tyr Ala Val Phe Gly
1385 1390 1395
Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala Gly Ile Asn Asp
1400 1405 1410
Val Ser Asn Phe Glu Thr Phe Gly Asn Ser Met Leu Cys Leu Phe
1415 1420 1425
Gln Val Ala Ile Phe Ala Gly Trp Asp Gly Met Leu Asp Ala Ile
1430 1435 1440
Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp Lys Ile Asn Pro
1445 1450 1455
Gly Thr Gln Val Arg Gly Asp Cys Gly Asn Pro Ser Val Gly Ile
1460 1465 1470
Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp Leu Ile Ile Val
1475 1480 1485
Asn Met Tyr Ile Val Val Val Met Glu Phe Leu Asn Ile Ala Ser
1490 1495 1500
Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp Phe Arg Lys Phe
1505 1510 1515
Phe Gln Val Trp Lys Arg Phe Asp Pro Asp Arg Thr Gln Tyr Ile
1520 1525 1530
Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu Asp Pro Pro
1535 1540 1545
Leu Phe Met Ala Lys Pro Asn Lys Gly Gln Leu Ile Ala Leu Asp
1550 1555 1560
Leu Pro Met Ala Val Gly Asp Arg Ile His Cys Leu Asp Ile Leu
1565 1570 1575
Leu Ala Phe Thr Lys Arg Val Met Gly Gln Asp Val Arg Met Glu
1580 1585 1590
Lys Val Val Ser Glu Ile Glu Ser Gly Phe Leu Leu Ala Asn Pro
1595 1600 1605
Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr Leu Lys Arg Lys
1610 1615 1620
Gln Glu Ala Val Ser Ala Thr Ile Ile Gln Arg Ala Tyr Lys Asn
1625 1630 1635
Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser Asp Ile His Met
1640 1645 1650
Ile Asp Gly Asp Arg Asp Val His Ala Thr Lys Glu Gly Ala Tyr
1655 1660 1665
Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln Ser Gln Ile
1670 1675 1680
<210> 2
<211> 1940
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_DI-DII_嵌合体
<400> 2
Met Ala Met Leu Pro Pro Pro Gly Pro Gln Ser Phe Val His Phe Thr
1 5 10 15
Lys Gln Ser Leu Ala Leu Ile Glu Gln Arg Ile Ala Glu Arg Lys Ser
20 25 30
Lys Glu Pro Lys Glu Glu Lys Lys Asp Asp Asp Glu Glu Ala Pro Lys
35 40 45
Pro Ser Ser Asp Leu Glu Ala Gly Lys Gln Leu Pro Phe Ile Tyr Gly
50 55 60
Asp Ile Pro Pro Gly Met Val Ser Glu Pro Leu Glu Asp Leu Asp Pro
65 70 75 80
Tyr Tyr Ala Asp Lys Lys Thr Phe Ile Val Leu Asn Lys Gly Lys Thr
85 90 95
Ile Phe Arg Phe Asn Ala Thr Pro Ala Leu Tyr Met Leu Ser Pro Phe
100 105 110
Ser Pro Leu Arg Arg Ile Ser Ile Lys Ile Leu Val His Ser Leu Phe
115 120 125
Ser Met Leu Ile Met Cys Thr Ile Leu Thr Asn Cys Ile Phe Met Thr
130 135 140
Met Asn Asn Pro Pro Asp Trp Thr Lys Asn Val Glu Tyr Thr Phe Thr
145 150 155 160
Gly Ile Tyr Thr Phe Glu Ser Leu Val Lys Ile Leu Ala Arg Gly Phe
165 170 175
Cys Val Gly Glu Phe Thr Phe Leu Arg Asp Pro Trp Asn Trp Leu Asp
180 185 190
Phe Val Val Ile Val Phe Ala Tyr Leu Thr Glu Phe Val Asn Leu Gly
195 200 205
Asn Val Ser Ala Leu Arg Thr Phe Arg Val Leu Arg Ala Leu Lys Thr
210 215 220
Ile Ser Val Ile Pro Gly Leu Lys Thr Ile Val Gly Ala Leu Ile Gln
225 230 235 240
Ser Val Lys Lys Leu Ser Asp Val Met Ile Leu Thr Val Phe Cys Leu
245 250 255
Ser Val Phe Ala Leu Ile Gly Leu Gln Leu Phe Met Gly Asn Leu Lys
260 265 270
His Lys Cys Phe Arg Asn Ser Leu Glu Asn Asn Glu Thr Leu Glu Ser
275 280 285
Ile Met Asn Thr Leu Glu Ser Glu Glu Asp Phe Arg Lys Tyr Phe Tyr
290 295 300
Tyr Leu Glu Gly Ser Lys Asp Ala Leu Leu Cys Gly Phe Ser Thr Asp
305 310 315 320
Ser Gly Gln Cys Pro Glu Gly Tyr Thr Cys Val Lys Ile Gly Arg Asn
325 330 335
Pro Asp Tyr Gly Tyr Thr Ser Phe Asp Thr Phe Ser Trp Ala Phe Leu
340 345 350
Ala Leu Phe Arg Leu Met Thr Gln Asp Tyr Trp Glu Asn Leu Tyr Gln
355 360 365
Gln Thr Leu Arg Ala Ala Gly Lys Thr Tyr Met Ile Phe Phe Val Val
370 375 380
Val Ile Phe Leu Gly Ser Phe Tyr Leu Ile Asn Leu Ile Leu Ala Val
385 390 395 400
Val Ala Met Ala Tyr Glu Glu Gln Asn Gln Ala Asn Ile Glu Glu Ala
405 410 415
Lys Gln Lys Glu Leu Glu Phe Gln Gln Met Leu Asp Arg Leu Lys Lys
420 425 430
Glu Gln Glu Glu Ala Glu Ala Ile Ala Ala Ala Ala Ala Glu Tyr Thr
435 440 445
Ser Ile Arg Arg Ser Arg Ile Met Gly Leu Ser Glu Ser Ser Ser Glu
450 455 460
Thr Ser Lys Leu Ser Ser Lys Ser Ala Lys Glu Arg Arg Asn Arg Arg
465 470 475 480
Lys Lys Lys Asn Gln Lys Lys Leu Ser Ser Gly Glu Glu Lys Gly Asp
485 490 495
Ala Glu Lys Leu Ser Lys Ser Glu Ser Glu Asp Ser Ile Arg Arg Lys
500 505 510
Ser Phe His Leu Gly Val Glu Gly His Arg Arg Ala His Glu Lys Arg
515 520 525
Leu Ser Thr Pro Asn Gln Ser Pro Leu Ser Ile Arg Gly Ser Leu Phe
530 535 540
Ser Ala Arg Arg Ser Ser Arg Thr Ser Leu Phe Ser Phe Lys Gly Arg
545 550 555 560
Gly Arg Asp Ile Gly Ser Glu Thr Glu Phe Ala Asp Asp Glu His Ser
565 570 575
Ile Phe Gly Asp Asn Glu Ser Arg Arg Gly Ser Leu Phe Val Pro His
580 585 590
Arg Pro Gln Glu Arg Arg Ser Ser Asn Ile Ser Gln Ala Ser Arg Ser
595 600 605
Pro Pro Met Leu Pro Val Asn Gly Lys Met His Ser Ala Val Asp Cys
610 615 620
Asn Gly Val Val Ser Leu Val Asp Gly Arg Ser Ala Leu Met Leu Pro
625 630 635 640
Asn Gly Gln Leu Leu Pro Glu Val Ile Ile Asp Lys Ala Thr Ser Asp
645 650 655
Asp Ser Gly Thr Thr Asn Gln Ile His Lys Lys Arg Arg Cys Ser Ser
660 665 670
Tyr Leu Leu Ser Glu Asp Met Leu Asn Asp Pro Asn Leu Arg Gln Arg
675 680 685
Ala Met Ser Arg Ala Ser Ile Leu Thr Asn Thr Val Glu Glu Leu Glu
690 695 700
Glu Ser Arg Gln Lys Cys Pro Pro Trp Trp Tyr Arg Phe Ala His Lys
705 710 715 720
Phe Leu Ile Trp Asn Cys Ser Pro Tyr Trp Ile Lys Phe Lys Lys Cys
725 730 735
Ile Tyr Phe Ile Val Met Asp Pro Phe Val Asp Leu Ala Ile Thr Ile
740 745 750
Cys Ile Val Leu Asn Thr Leu Phe Met Ala Met Glu His His Pro Met
755 760 765
Thr Glu Glu Phe Lys Asn Val Leu Ala Ile Gly Asn Leu Val Phe Thr
770 775 780
Gly Ile Phe Ala Ala Glu Met Val Leu Lys Leu Ile Ala Met Asp Pro
785 790 795 800
Tyr Glu Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Leu Ile Val
805 810 815
Thr Leu Ser Leu Val Glu Leu Phe Leu Ala Asp Val Glu Gly Leu Ser
820 825 830
Val Leu Arg Ser Phe Arg Leu Leu Arg Val Phe Lys Leu Ala Lys Ser
835 840 845
Trp Pro Thr Leu Asn Met Leu Ile Lys Ile Ile Gly Asn Ser Val Gly
850 855 860
Ala Leu Gly Asn Leu Thr Leu Val Leu Ala Ile Ile Val Phe Ile Phe
865 870 875 880
Ala Val Val Gly Met Gln Leu Phe Gly Lys Ser Tyr Lys Glu Cys Val
885 890 895
Cys Lys Ile Asn Asp Asp Cys Thr Leu Pro Arg Trp His Met Asn Asp
900 905 910
Phe Phe His Ser Phe Leu Ile Val Phe Arg Val Leu Cys Gly Glu Trp
915 920 925
Ile Glu Thr Met Trp Asp Cys Met Glu Val Ala Gly Gln Ala Met Cys
930 935 940
Leu Ile Val Tyr Met Met Val Met Val Ile Gly Asn Leu Val Val Leu
945 950 955 960
Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe Ser Ser Asp Asn Leu
965 970 975
Thr Ala Ile Glu Glu Asp Pro Asp Ala Asn Asn Leu Gln Ile Ala Val
980 985 990
Thr Arg Ile Lys Lys Gly Ile Asn Tyr Val Lys Gln Thr Leu Arg Glu
995 1000 1005
Phe Ile Leu Lys Ala Phe Ser Lys Lys Pro Lys Ile Ser Arg Glu
1010 1015 1020
Ile Arg Gln Ala Glu Asp Leu Asn Thr Lys Lys Glu Asn Tyr Ile
1025 1030 1035
Ser Asn His Thr Leu Ala Glu Met Ser Lys Gly His Asn Phe Leu
1040 1045 1050
Lys Glu Lys Asp Lys Ile Ser Gly Phe Gly Ser Ser Val Asp Lys
1055 1060 1065
His Leu Met Glu Asp Ser Asp Gly Gln Ser Phe Ile His Asn Pro
1070 1075 1080
Ser Leu Thr Val Thr Val Pro Ile Ala Pro Gly Glu Ser Asp Leu
1085 1090 1095
Glu Asn Met Asn Ala Glu Glu Leu Ser Ser Asp Ser Asp Ser Glu
1100 1105 1110
Tyr Ser Lys Val Arg Leu Asn Arg Ser Ser Ser Ser Glu Cys Ser
1115 1120 1125
Thr Val Asp Asn Pro Leu Pro Gly Glu Gly Glu Glu Ala Glu Ala
1130 1135 1140
Glu Pro Met Asn Ser Asp Glu Pro Glu Ala Cys Phe Thr Asp Gly
1145 1150 1155
Cys Val Trp Arg Phe Ser Cys Cys Gln Val Asn Ile Glu Ser Gly
1160 1165 1170
Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys Thr Cys Cys Lys Ile
1175 1180 1185
Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly Leu Val Thr Leu
1190 1195 1200
Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln
1205 1210 1215
Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe
1220 1225 1230
Thr Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr
1235 1240 1245
Gly Phe Lys Ala Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe
1250 1255 1260
Val Val Val Ile Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg
1265 1270 1275
Glu Glu Leu Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu
1280 1285 1290
Arg Val Leu Ser Gln Phe Glu Arg Met Lys Val Val Val Arg Ala
1295 1300 1305
Leu Ile Lys Thr Thr Leu Pro Thr Leu Asn Val Phe Leu Val Cys
1310 1315 1320
Leu Met Ile Trp Leu Ile Phe Ser Ile Met Gly Val Asp Leu Phe
1325 1330 1335
Ala Gly Arg Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg
1340 1345 1350
Phe Pro Ser Ser Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu
1355 1360 1365
Leu Phe Asn Glu Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe
1370 1375 1380
Asp Asn Val Gly Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr
1385 1390 1395
Phe Asn Gly Trp Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val
1400 1405 1410
Ala Val Asn Ile Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr
1415 1420 1425
Cys Tyr Phe Ile Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu
1430 1435 1440
Ser Met Leu Ile Thr Val Ile Ile Asp Asn Phe Asn Lys His Lys
1445 1450 1455
Ile Lys Leu Gly Gly Ser Asn Ile Phe Ile Thr Val Lys Gln Arg
1460 1465 1470
Lys Gln Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser Gln
1475 1480 1485
Arg Pro Val Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe Ile Phe
1490 1495 1500
Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val Met Val Leu
1505 1510 1515
Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp Val Gln
1520 1525 1530
Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile Phe
1535 1540 1545
Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile Ala Phe
1550 1555 1560
Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe Met
1565 1570 1575
Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met Thr Val
1580 1585 1590
Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile Leu Leu
1595 1600 1605
Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro Lys Val
1610 1615 1620
Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro Ala Leu
1625 1630 1635
Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile Tyr Ala
1640 1645 1650
Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala Gly
1655 1660 1665
Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser Met Leu
1670 1675 1680
Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly Met Leu
1685 1690 1695
Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp Lys
1700 1705 1710
Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn Pro Ser
1715 1720 1725
Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp Leu
1730 1735 1740
Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu Phe Leu Asn
1745 1750 1755
Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp Phe
1760 1765 1770
Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro Asp Arg Thr
1775 1780 1785
Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu
1790 1795 1800
Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly Gln Leu Ile
1805 1810 1815
Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile His Cys Leu
1820 1825 1830
Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly Gln Asp Val
1835 1840 1845
Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly Phe Leu Leu
1850 1855 1860
Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr Leu
1865 1870 1875
Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile Gln Arg Ala
1880 1885 1890
Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser Asp
1895 1900 1905
Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala Thr Lys Glu
1910 1915 1920
Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln Ser
1925 1930 1935
Gln Ile
1940
<210> 3
<211> 1730
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_DIII-DIV_嵌合体
<400> 3
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys Thr Cys Tyr
915 920 925
Lys Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile Val Leu Met Ile
930 935 940
Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Arg
945 950 955 960
Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala Tyr Gly Tyr
980 985 990
Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val
995 1000 1005
Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu Gly Tyr Ser
1010 1015 1020
Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg
1025 1030 1035
Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val
1040 1045 1050
Asn Ala Leu Ile Gly Ala Ile Pro Ser Ile Met Asn Val Leu Leu
1055 1060 1065
Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile Met Gly Val Asn
1070 1075 1080
Leu Phe Ala Gly Lys Phe Tyr Glu Cys Ile Asn Thr Thr Asp Gly
1085 1090 1095
Ser Arg Phe Pro Ala Ser Gln Val Pro Asn Arg Ser Glu Cys Phe
1100 1105 1110
Ala Leu Met Asn Val Ser Gln Asn Val Arg Trp Lys Asn Leu Lys
1115 1120 1125
Val Asn Phe Asp Asn Val Gly Leu Gly Tyr Leu Ser Leu Leu Gln
1130 1135 1140
Val Ala Thr Phe Lys Gly Trp Thr Ile Ile Met Tyr Ala Ala Val
1145 1150 1155
Asp Ser Val Asn Val Asp Lys Gln Pro Lys Tyr Glu Tyr Ser Leu
1160 1165 1170
Tyr Met Tyr Ile Tyr Phe Val Val Phe Ile Ile Phe Gly Ser Phe
1175 1180 1185
Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe Asn
1190 1195 1200
Gln Gln Lys Lys Lys Leu Gly Gly Gln Asp Ile Phe Met Thr Glu
1205 1210 1215
Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys
1220 1225 1230
Lys Pro Gln Lys Pro Ile Pro Arg Pro Gly Asn Lys Ile Gln Gly
1235 1240 1245
Cys Ile Phe Asp Leu Val Thr Asn Gln Ala Phe Asp Ile Ser Ile
1250 1255 1260
Met Val Leu Ile Cys Leu Asn Met Val Thr Met Met Val Glu Lys
1265 1270 1275
Glu Gly Gln Ser Gln His Met Thr Glu Val Leu Tyr Trp Ile Asn
1280 1285 1290
Val Val Phe Ile Ile Leu Phe Thr Gly Glu Cys Val Leu Lys Leu
1295 1300 1305
Ile Ser Leu Arg His Tyr Tyr Phe Thr Val Gly Trp Asn Ile Phe
1310 1315 1320
Asp Phe Val Val Val Ile Ile Ser Ile Val Gly Met Phe Leu Ala
1325 1330 1335
Asp Leu Ile Glu Thr Tyr Phe Val Ser Pro Thr Leu Phe Arg Val
1340 1345 1350
Ile Arg Leu Ala Arg Ile Gly Arg Ile Leu Arg Leu Val Lys Gly
1355 1360 1365
Ala Lys Gly Ile Arg Thr Leu Leu Phe Ala Leu Met Met Ser Leu
1370 1375 1380
Pro Ala Leu Phe Asn Ile Gly Leu Leu Leu Phe Leu Val Met Phe
1385 1390 1395
Ile Tyr Ala Ile Phe Gly Met Ser Asn Phe Ala Tyr Val Lys Lys
1400 1405 1410
Glu Asp Gly Ile Asn Asp Met Phe Asn Phe Glu Thr Phe Gly Asn
1415 1420 1425
Ser Met Ile Cys Leu Phe Gln Ile Thr Thr Ser Ala Gly Trp Asp
1430 1435 1440
Gly Leu Leu Ala Pro Ile Leu Asn Ser Lys Pro Pro Asp Cys Asp
1445 1450 1455
Pro Lys Lys Val His Pro Gly Ser Ser Val Glu Gly Asp Cys Gly
1460 1465 1470
Asn Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Ile Ile
1475 1480 1485
Ser Phe Leu Val Val Val Asn Met Tyr Ile Ala Val Ile Leu Glu
1490 1495 1500
Asn Phe Ser Val Ala Thr Glu Glu Ser Thr Glu Pro Leu Ser Glu
1505 1510 1515
Asp Asp Phe Glu Met Phe Tyr Glu Val Trp Glu Lys Phe Asp Pro
1520 1525 1530
Asp Ala Thr Gln Phe Ile Glu Phe Ser Lys Leu Ser Asp Phe Ala
1535 1540 1545
Ala Ala Leu Asp Pro Pro Leu Leu Ile Ala Lys Pro Asn Lys Val
1550 1555 1560
Gln Leu Ile Ala Met Asp Leu Pro Met Val Ser Gly Asp Arg Ile
1565 1570 1575
His Cys Leu Asp Ile Leu Phe Ala Phe Thr Lys Arg Val Leu Gly
1580 1585 1590
Glu Ser Gly Glu Met Asp Ser Leu Arg Ser Gln Met Glu Glu Arg
1595 1600 1605
Phe Met Ser Ala Asn Pro Ser Lys Val Ser Tyr Glu Pro Ile Thr
1610 1615 1620
Thr Thr Leu Lys Arg Lys Gln Glu Asp Val Ser Ala Thr Val Ile
1625 1630 1635
Gln Arg Ala Tyr Arg Arg Tyr Arg Leu Arg Gln Asn Val Lys Asn
1640 1645 1650
Ile Ser Ser Ile Tyr Ile Lys Asp Gly Asp Arg Asp Asp Asp Leu
1655 1660 1665
Leu Asn Lys Lys Asp Met Ala Phe Asp Asn Val Asn Glu Asn Ser
1670 1675 1680
Ser Pro Glu Lys Thr Asp Ala Thr Ser Ser Thr Thr Ser Pro Pro
1685 1690 1695
Ser Tyr Asp Ser Val Thr Lys Pro Asp Lys Glu Lys Tyr Glu Gln
1700 1705 1710
Asp Arg Thr Glu Lys Glu Asp Lys Gly Lys Asp Ser Lys Glu Ser
1715 1720 1725
Lys Lys
1730
<210> 4
<211> 1707
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_DII-DIII_嵌合体
<400> 4
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Tyr Trp Ile Lys Phe Lys Lys
485 490 495
Cys Ile Tyr Phe Ile Val Met Asp Pro Phe Val Asp Leu Ala Ile Thr
500 505 510
Ile Cys Ile Val Leu Asn Thr Leu Phe Met Ala Met Glu His His Pro
515 520 525
Met Thr Glu Glu Phe Lys Asn Val Leu Ala Ile Gly Asn Leu Val Phe
530 535 540
Thr Gly Ile Phe Ala Ala Glu Met Val Leu Lys Leu Ile Ala Met Asp
545 550 555 560
Pro Tyr Glu Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Leu Ile
565 570 575
Val Thr Leu Ser Leu Val Glu Leu Phe Leu Ala Asp Val Glu Gly Leu
580 585 590
Ser Val Leu Arg Ser Phe Arg Leu Leu Arg Val Phe Lys Leu Ala Lys
595 600 605
Ser Trp Pro Thr Leu Asn Met Leu Ile Lys Ile Ile Gly Asn Ser Val
610 615 620
Gly Ala Leu Gly Asn Leu Thr Leu Val Leu Ala Ile Ile Val Phe Ile
625 630 635 640
Phe Ala Val Val Gly Met Gln Leu Phe Gly Lys Ser Tyr Lys Glu Cys
645 650 655
Val Cys Lys Ile Asn Asp Asp Cys Thr Leu Pro Arg Trp His Met Asn
660 665 670
Asp Phe Phe His Ser Phe Leu Ile Val Phe Arg Val Leu Cys Gly Glu
675 680 685
Trp Ile Glu Thr Met Trp Asp Cys Met Glu Val Ala Gly Gln Ala Met
690 695 700
Cys Leu Ile Val Tyr Met Met Val Met Val Ile Gly Asn Leu Val Val
705 710 715 720
Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe Ser Ser Asp Asn
725 730 735
Leu Thr Ala Ile Glu Glu Asp Pro Asp Ala Asn Asn Leu Gln Ile Ala
740 745 750
Val Thr Arg Ile Lys Lys Gly Ile Asn Tyr Val Lys Gln Thr Leu Arg
755 760 765
Glu Phe Ile Leu Lys Ala Phe Ser Lys Lys Pro Lys Ile Ser Arg Glu
770 775 780
Ile Arg Gln Ala Glu Asp Leu Asn Thr Lys Lys Glu Asn Tyr Ile Ser
785 790 795 800
Asn His Thr Leu Ala Glu Met Ser Lys Gly His Asn Phe Leu Lys Glu
805 810 815
Lys Asp Lys Ile Ser Gly Phe Gly Ser Ser Val Asp Lys His Leu Met
820 825 830
Glu Asp Ser Asp Gly Gln Ser Phe Ile His Asn Pro Ser Leu Thr Val
835 840 845
Thr Val Pro Ile Ala Pro Gly Glu Ser Asp Leu Glu Asn Met Asn Ala
850 855 860
Glu Glu Leu Ser Ser Asp Ser Asp Ser Glu Tyr Ser Lys Val Arg Leu
865 870 875 880
Asn Arg Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Asn Pro Leu Pro
885 890 895
Gly Glu Gly Glu Glu Ala Glu Ala Glu Pro Met Asn Ser Asp Glu Pro
900 905 910
Glu Ala Cys Phe Thr Asp Gly Cys Val Trp Arg Phe Ser Cys Cys Gln
915 920 925
Val Asn Ile Glu Ser Gly Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys
930 935 940
Thr Cys Tyr Lys Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile Val
945 950 955 960
Leu Met Ile Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr
965 970 975
Ile Glu Arg Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys
980 985 990
Ile Phe Thr Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala
995 1000 1005
Tyr Gly Tyr Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp
1010 1015 1020
Phe Leu Ile Val Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr
1025 1030 1035
Leu Gly Tyr Ser Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu
1040 1045 1050
Arg Ala Leu Arg Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met
1055 1060 1065
Arg Val Val Val Asn Ala Leu Ile Gly Ala Ile Pro Ser Ile Met
1070 1075 1080
Asn Val Leu Leu Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile
1085 1090 1095
Met Gly Val Asn Leu Phe Ala Gly Lys Phe Tyr Glu Cys Ile Asn
1100 1105 1110
Thr Thr Asp Gly Ser Arg Phe Pro Ala Ser Gln Val Pro Asn Arg
1115 1120 1125
Ser Glu Cys Phe Ala Leu Met Asn Val Ser Gln Asn Val Arg Trp
1130 1135 1140
Lys Asn Leu Lys Val Asn Phe Asp Asn Val Gly Leu Gly Tyr Leu
1145 1150 1155
Ser Leu Leu Gln Val Ala Thr Phe Lys Gly Trp Thr Ile Ile Met
1160 1165 1170
Tyr Ala Ala Val Asp Ser Val Asn Val Asp Lys Gln Pro Lys Tyr
1175 1180 1185
Glu Tyr Ser Leu Tyr Met Tyr Ile Tyr Phe Val Val Phe Ile Ile
1190 1195 1200
Phe Gly Ser Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile
1205 1210 1215
Asp Asn Phe Asn Gln Gln Lys Lys Lys Leu Gly Gly Gln Asp Ile
1220 1225 1230
Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys
1235 1240 1245
Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg Pro Gly Asn
1250 1255 1260
Lys Leu Gln Gly Phe Ile Phe Asp Val Val Thr Ser Gln Ala Phe
1265 1270 1275
Asn Val Ile Val Met Val Leu Ile Cys Phe Gln Ala Ile Ala Met
1280 1285 1290
Met Ile Asp Thr Asp Val Gln Ser Leu Gln Met Ser Ile Ala Leu
1295 1300 1305
Tyr Trp Ile Asn Ser Ile Phe Val Met Leu Tyr Thr Met Glu Cys
1310 1315 1320
Ile Leu Lys Leu Ile Ala Phe Arg Cys Phe Tyr Phe Thr Ile Ala
1325 1330 1335
Trp Asn Ile Phe Asp Phe Met Val Val Ile Phe Ser Ile Thr Gly
1340 1345 1350
Leu Cys Leu Pro Met Thr Val Gly Ser Tyr Leu Val Pro Pro Ser
1355 1360 1365
Leu Val Gln Leu Ile Leu Leu Ser Arg Ile Ile His Met Leu Arg
1370 1375 1380
Leu Gly Lys Gly Pro Lys Val Phe His Asn Leu Met Leu Pro Leu
1385 1390 1395
Met Leu Ser Leu Pro Ala Leu Leu Asn Ile Ile Leu Leu Ile Phe
1400 1405 1410
Leu Val Met Phe Ile Tyr Ala Val Phe Gly Met Tyr Asn Phe Ala
1415 1420 1425
Tyr Val Lys Lys Glu Ala Gly Ile Asn Asp Val Ser Asn Phe Glu
1430 1435 1440
Thr Phe Gly Asn Ser Met Leu Cys Leu Phe Gln Val Ala Ile Phe
1445 1450 1455
Ala Gly Trp Asp Gly Met Leu Asp Ala Ile Phe Asn Ser Lys Trp
1460 1465 1470
Ser Asp Cys Asp Pro Asp Lys Ile Asn Pro Gly Thr Gln Val Arg
1475 1480 1485
Gly Asp Cys Gly Asn Pro Ser Val Gly Ile Phe Tyr Phe Val Ser
1490 1495 1500
Tyr Ile Leu Ile Ser Trp Leu Ile Ile Val Asn Met Tyr Ile Val
1505 1510 1515
Val Val Met Glu Phe Leu Asn Ile Ala Ser Lys Lys Lys Asn Lys
1520 1525 1530
Thr Leu Ser Glu Asp Asp Phe Arg Lys Phe Phe Gln Val Trp Lys
1535 1540 1545
Arg Phe Asp Pro Asp Arg Thr Gln Tyr Ile Asp Ser Ser Lys Leu
1550 1555 1560
Ser Asp Phe Ala Ala Ala Leu Asp Pro Pro Leu Phe Met Ala Lys
1565 1570 1575
Pro Asn Lys Gly Gln Leu Ile Ala Leu Asp Leu Pro Met Ala Val
1580 1585 1590
Gly Asp Arg Ile His Cys Leu Asp Ile Leu Leu Ala Phe Thr Lys
1595 1600 1605
Arg Val Met Gly Gln Asp Val Arg Met Glu Lys Val Val Ser Glu
1610 1615 1620
Ile Glu Ser Gly Phe Leu Leu Ala Asn Pro Phe Lys Ile Thr Cys
1625 1630 1635
Glu Pro Ile Thr Thr Thr Leu Lys Arg Lys Gln Glu Ala Val Ser
1640 1645 1650
Ala Thr Ile Ile Gln Arg Ala Tyr Lys Asn Tyr Arg Leu Arg Arg
1655 1660 1665
Asn Asp Lys Asn Thr Ser Asp Ile His Met Ile Asp Gly Asp Arg
1670 1675 1680
Asp Val His Ala Thr Lys Glu Gly Ala Tyr Phe Asp Lys Ala Lys
1685 1690 1695
Glu Lys Ser Pro Ile Gln Ser Gln Ile
1700 1705
<210> 5
<211> 1963
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_DI-DIV_嵌合体
<400> 5
Met Ala Met Leu Pro Pro Pro Gly Pro Gln Ser Phe Val His Phe Thr
1 5 10 15
Lys Gln Ser Leu Ala Leu Ile Glu Gln Arg Ile Ala Glu Arg Lys Ser
20 25 30
Lys Glu Pro Lys Glu Glu Lys Lys Asp Asp Asp Glu Glu Ala Pro Lys
35 40 45
Pro Ser Ser Asp Leu Glu Ala Gly Lys Gln Leu Pro Phe Ile Tyr Gly
50 55 60
Asp Ile Pro Pro Gly Met Val Ser Glu Pro Leu Glu Asp Leu Asp Pro
65 70 75 80
Tyr Tyr Ala Asp Lys Lys Thr Phe Ile Val Leu Asn Lys Gly Lys Thr
85 90 95
Ile Phe Arg Phe Asn Ala Thr Pro Ala Leu Tyr Met Leu Ser Pro Phe
100 105 110
Ser Pro Leu Arg Arg Ile Ser Ile Lys Ile Leu Val His Ser Leu Phe
115 120 125
Ser Met Leu Ile Met Cys Thr Ile Leu Thr Asn Cys Ile Phe Met Thr
130 135 140
Met Asn Asn Pro Pro Asp Trp Thr Lys Asn Val Glu Tyr Thr Phe Thr
145 150 155 160
Gly Ile Tyr Thr Phe Glu Ser Leu Val Lys Ile Leu Ala Arg Gly Phe
165 170 175
Cys Val Gly Glu Phe Thr Phe Leu Arg Asp Pro Trp Asn Trp Leu Asp
180 185 190
Phe Val Val Ile Val Phe Ala Tyr Leu Thr Glu Phe Val Asn Leu Gly
195 200 205
Asn Val Ser Ala Leu Arg Thr Phe Arg Val Leu Arg Ala Leu Lys Thr
210 215 220
Ile Ser Val Ile Pro Gly Leu Lys Thr Ile Val Gly Ala Leu Ile Gln
225 230 235 240
Ser Val Lys Lys Leu Ser Asp Val Met Ile Leu Thr Val Phe Cys Leu
245 250 255
Ser Val Phe Ala Leu Ile Gly Leu Gln Leu Phe Met Gly Asn Leu Lys
260 265 270
His Lys Cys Phe Arg Asn Ser Leu Glu Asn Asn Glu Thr Leu Glu Ser
275 280 285
Ile Met Asn Thr Leu Glu Ser Glu Glu Asp Phe Arg Lys Tyr Phe Tyr
290 295 300
Tyr Leu Glu Gly Ser Lys Asp Ala Leu Leu Cys Gly Phe Ser Thr Asp
305 310 315 320
Ser Gly Gln Cys Pro Glu Gly Tyr Thr Cys Val Lys Ile Gly Arg Asn
325 330 335
Pro Asp Tyr Gly Tyr Thr Ser Phe Asp Thr Phe Ser Trp Ala Phe Leu
340 345 350
Ala Leu Phe Arg Leu Met Thr Gln Asp Tyr Trp Glu Asn Leu Tyr Gln
355 360 365
Gln Thr Leu Arg Ala Ala Gly Lys Thr Tyr Met Ile Phe Phe Val Val
370 375 380
Val Ile Phe Leu Gly Ser Phe Tyr Leu Ile Asn Leu Ile Leu Ala Val
385 390 395 400
Val Ala Met Ala Tyr Glu Glu Gln Asn Gln Ala Asn Ile Glu Glu Ala
405 410 415
Lys Gln Lys Glu Leu Glu Phe Gln Gln Met Leu Asp Arg Leu Lys Lys
420 425 430
Glu Gln Glu Glu Ala Glu Ala Ile Ala Ala Ala Ala Ala Glu Tyr Thr
435 440 445
Ser Ile Arg Arg Ser Arg Ile Met Gly Leu Ser Glu Ser Ser Ser Glu
450 455 460
Thr Ser Lys Leu Ser Ser Lys Ser Ala Lys Glu Arg Arg Asn Arg Arg
465 470 475 480
Lys Lys Lys Asn Gln Lys Lys Leu Ser Ser Gly Glu Glu Lys Gly Asp
485 490 495
Ala Glu Lys Leu Ser Lys Ser Glu Ser Glu Asp Ser Ile Arg Arg Lys
500 505 510
Ser Phe His Leu Gly Val Glu Gly His Arg Arg Ala His Glu Lys Arg
515 520 525
Leu Ser Thr Pro Asn Gln Ser Pro Leu Ser Ile Arg Gly Ser Leu Phe
530 535 540
Ser Ala Arg Arg Ser Ser Arg Thr Ser Leu Phe Ser Phe Lys Gly Arg
545 550 555 560
Gly Arg Asp Ile Gly Ser Glu Thr Glu Phe Ala Asp Asp Glu His Ser
565 570 575
Ile Phe Gly Asp Asn Glu Ser Arg Arg Gly Ser Leu Phe Val Pro His
580 585 590
Arg Pro Gln Glu Arg Arg Ser Ser Asn Ile Ser Gln Ala Ser Arg Ser
595 600 605
Pro Pro Met Leu Pro Val Asn Gly Lys Met His Ser Ala Val Asp Cys
610 615 620
Asn Gly Val Val Ser Leu Val Asp Gly Arg Ser Ala Leu Met Leu Pro
625 630 635 640
Asn Gly Gln Leu Leu Pro Glu Val Ile Ile Asp Lys Ala Thr Ser Asp
645 650 655
Asp Ser Gly Thr Thr Asn Gln Ile His Lys Lys Arg Arg Cys Ser Ser
660 665 670
Tyr Leu Leu Ser Glu Asp Met Leu Asn Asp Pro Asn Leu Arg Gln Arg
675 680 685
Ala Met Ser Arg Ala Ser Ile Leu Thr Asn Thr Val Glu Glu Leu Glu
690 695 700
Glu Ser Arg Gln Lys Cys Pro Pro Trp Trp Tyr Arg Phe Ala His Lys
705 710 715 720
Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu Phe
725 730 735
Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile Ile
740 745 750
Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro Met
755 760 765
Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe Ile
770 775 780
Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His Pro
785 790 795 800
Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile Val
805 810 815
Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met Ala
820 825 830
Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys Tyr
835 840 845
Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp Val
850 855 860
Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe Ser
865 870 875 880
Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe Val
885 890 895
Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His Asp
900 905 910
Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu Trp
915 920 925
Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp Cys
930 935 940
Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val Leu
945 950 955 960
Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp Val
965 970 975
Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val Ala
980 985 990
Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys Lys
995 1000 1005
Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
1010 1015 1020
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser
1025 1030 1035
Asn Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr
1040 1045 1050
Glu Lys Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser
1055 1060 1065
Pro Ser Val Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp
1070 1075 1080
Ile Glu Asn Leu Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp
1085 1090 1095
Gly Gly Ser Lys Glu Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys
1100 1105 1110
Ser Thr Val Asp Ile Ala Ile Ser Glu Glu Glu Glu Met Phe Tyr
1115 1120 1125
Gly Gly Glu Arg Ser Lys His Leu Lys Asn Gly Cys Arg Arg Gly
1130 1135 1140
Ser Ser Leu Gly Gln Ile Ser Gly Ala Ser Lys Lys Gly Lys Ile
1145 1150 1155
Trp Gln Asn Ile Arg Lys Thr Cys Cys Lys Ile Val Glu Asn Asn
1160 1165 1170
Trp Phe Lys Cys Phe Ile Gly Leu Val Thr Leu Leu Ser Thr Gly
1175 1180 1185
Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln Arg Lys Thr Ile
1190 1195 1200
Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr Tyr Ile Phe
1205 1210 1215
Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe Lys Ala
1220 1225 1230
Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val Ile
1235 1240 1245
Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg Glu Glu Leu Lys
1250 1255 1260
Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu Ser
1265 1270 1275
Gln Phe Glu Arg Met Lys Val Val Val Arg Ala Leu Ile Lys Thr
1280 1285 1290
Thr Leu Pro Thr Leu Asn Val Phe Leu Val Cys Leu Met Ile Trp
1295 1300 1305
Leu Ile Phe Ser Ile Met Gly Val Asp Leu Phe Ala Gly Arg Phe
1310 1315 1320
Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg Phe Pro Ser Ser
1325 1330 1335
Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu Leu Phe Asn Glu
1340 1345 1350
Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe Asp Asn Val Gly
1355 1360 1365
Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr Phe Asn Gly Trp
1370 1375 1380
Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val Ala Val Asn Ile
1385 1390 1395
Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr Cys Tyr Phe Ile
1400 1405 1410
Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu Ser Met Leu Ile
1415 1420 1425
Thr Val Ile Ile Asp Asn Phe Asn Lys His Lys Ile Lys Leu Gly
1430 1435 1440
Gly Ser Asn Ile Phe Ile Thr Val Lys Gln Arg Lys Gln Tyr Arg
1445 1450 1455
Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser Gln Arg Pro Val Pro
1460 1465 1470
Arg Pro Leu Asn Lys Ile Gln Gly Cys Ile Phe Asp Leu Val Thr
1475 1480 1485
Asn Gln Ala Phe Asp Ile Ser Ile Met Val Leu Ile Cys Leu Asn
1490 1495 1500
Met Val Thr Met Met Val Glu Lys Glu Gly Gln Ser Gln His Met
1505 1510 1515
Thr Glu Val Leu Tyr Trp Ile Asn Val Val Phe Ile Ile Leu Phe
1520 1525 1530
Thr Gly Glu Cys Val Leu Lys Leu Ile Ser Leu Arg His Tyr Tyr
1535 1540 1545
Phe Thr Val Gly Trp Asn Ile Phe Asp Phe Val Val Val Ile Ile
1550 1555 1560
Ser Ile Val Gly Met Phe Leu Ala Asp Leu Ile Glu Thr Tyr Phe
1565 1570 1575
Val Ser Pro Thr Leu Phe Arg Val Ile Arg Leu Ala Arg Ile Gly
1580 1585 1590
Arg Ile Leu Arg Leu Val Lys Gly Ala Lys Gly Ile Arg Thr Leu
1595 1600 1605
Leu Phe Ala Leu Met Met Ser Leu Pro Ala Leu Phe Asn Ile Gly
1610 1615 1620
Leu Leu Leu Phe Leu Val Met Phe Ile Tyr Ala Ile Phe Gly Met
1625 1630 1635
Ser Asn Phe Ala Tyr Val Lys Lys Glu Asp Gly Ile Asn Asp Met
1640 1645 1650
Phe Asn Phe Glu Thr Phe Gly Asn Ser Met Ile Cys Leu Phe Gln
1655 1660 1665
Ile Thr Thr Ser Ala Gly Trp Asp Gly Leu Leu Ala Pro Ile Leu
1670 1675 1680
Asn Ser Lys Pro Pro Asp Cys Asp Pro Lys Lys Val His Pro Gly
1685 1690 1695
Ser Ser Val Glu Gly Asp Cys Gly Asn Pro Ser Val Gly Ile Phe
1700 1705 1710
Tyr Phe Val Ser Tyr Ile Ile Ile Ser Phe Leu Val Val Val Asn
1715 1720 1725
Met Tyr Ile Ala Val Ile Leu Glu Asn Phe Ser Val Ala Thr Glu
1730 1735 1740
Glu Ser Thr Glu Pro Leu Ser Glu Asp Asp Phe Glu Met Phe Tyr
1745 1750 1755
Glu Val Trp Glu Lys Phe Asp Pro Asp Ala Thr Gln Phe Ile Glu
1760 1765 1770
Phe Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu Asp Pro Pro Leu
1775 1780 1785
Leu Ile Ala Lys Pro Asn Lys Val Gln Leu Ile Ala Met Asp Leu
1790 1795 1800
Pro Met Val Ser Gly Asp Arg Ile His Cys Leu Asp Ile Leu Phe
1805 1810 1815
Ala Phe Thr Lys Arg Val Leu Gly Glu Ser Gly Glu Met Asp Ser
1820 1825 1830
Leu Arg Ser Gln Met Glu Glu Arg Phe Met Ser Ala Asn Pro Ser
1835 1840 1845
Lys Val Ser Tyr Glu Pro Ile Thr Thr Thr Leu Lys Arg Lys Gln
1850 1855 1860
Glu Asp Val Ser Ala Thr Val Ile Gln Arg Ala Tyr Arg Arg Tyr
1865 1870 1875
Arg Leu Arg Gln Asn Val Lys Asn Ile Ser Ser Ile Tyr Ile Lys
1880 1885 1890
Asp Gly Asp Arg Asp Asp Asp Leu Leu Asn Lys Lys Asp Met Ala
1895 1900 1905
Phe Asp Asn Val Asn Glu Asn Ser Ser Pro Glu Lys Thr Asp Ala
1910 1915 1920
Thr Ser Ser Thr Thr Ser Pro Pro Ser Tyr Asp Ser Val Thr Lys
1925 1930 1935
Pro Asp Lys Glu Lys Tyr Glu Gln Asp Arg Thr Glu Lys Glu Asp
1940 1945 1950
Lys Gly Lys Asp Ser Lys Glu Ser Lys Lys
1955 1960
<210> 6
<211> 1921
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX-hNaV1.7_DI_嵌合体
<400> 6
Met Ala Met Leu Pro Pro Pro Gly Pro Gln Ser Phe Val His Phe Thr
1 5 10 15
Lys Gln Ser Leu Ala Leu Ile Glu Gln Arg Ile Ala Glu Arg Lys Ser
20 25 30
Lys Glu Pro Lys Glu Glu Lys Lys Asp Asp Asp Glu Glu Ala Pro Lys
35 40 45
Pro Ser Ser Asp Leu Glu Ala Gly Lys Gln Leu Pro Phe Ile Tyr Gly
50 55 60
Asp Ile Pro Pro Gly Met Val Ser Glu Pro Leu Glu Asp Leu Asp Pro
65 70 75 80
Tyr Tyr Ala Asp Lys Lys Thr Phe Ile Val Leu Asn Lys Gly Lys Thr
85 90 95
Ile Phe Arg Phe Asn Ala Thr Pro Ala Leu Tyr Met Leu Ser Pro Phe
100 105 110
Ser Pro Leu Arg Arg Ile Ser Ile Lys Ile Leu Val His Ser Leu Phe
115 120 125
Ser Met Leu Ile Met Cys Thr Ile Leu Thr Asn Cys Ile Phe Met Thr
130 135 140
Met Asn Asn Pro Pro Asp Trp Thr Lys Asn Val Glu Tyr Thr Phe Thr
145 150 155 160
Gly Ile Tyr Thr Phe Glu Ser Leu Val Lys Ile Leu Ala Arg Gly Phe
165 170 175
Cys Val Gly Glu Phe Thr Phe Leu Arg Asp Pro Trp Asn Trp Leu Asp
180 185 190
Phe Val Val Ile Val Phe Ala Tyr Leu Thr Glu Phe Val Asn Leu Gly
195 200 205
Asn Val Ser Ala Leu Arg Thr Phe Arg Val Leu Arg Ala Leu Lys Thr
210 215 220
Ile Ser Val Ile Pro Gly Leu Lys Thr Ile Val Gly Ala Leu Ile Gln
225 230 235 240
Ser Val Lys Lys Leu Ser Asp Val Met Ile Leu Thr Val Phe Cys Leu
245 250 255
Ser Val Phe Ala Leu Ile Gly Leu Gln Leu Phe Met Gly Asn Leu Lys
260 265 270
His Lys Cys Phe Arg Asn Ser Leu Glu Asn Asn Glu Thr Leu Glu Ser
275 280 285
Ile Met Asn Thr Leu Glu Ser Glu Glu Asp Phe Arg Lys Tyr Phe Tyr
290 295 300
Tyr Leu Glu Gly Ser Lys Asp Ala Leu Leu Cys Gly Phe Ser Thr Asp
305 310 315 320
Ser Gly Gln Cys Pro Glu Gly Tyr Thr Cys Val Lys Ile Gly Arg Asn
325 330 335
Pro Asp Tyr Gly Tyr Thr Ser Phe Asp Thr Phe Ser Trp Ala Phe Leu
340 345 350
Ala Leu Phe Arg Leu Met Thr Gln Asp Tyr Trp Glu Asn Leu Tyr Gln
355 360 365
Gln Thr Leu Arg Ala Ala Gly Lys Thr Tyr Met Ile Phe Phe Val Val
370 375 380
Val Ile Phe Leu Gly Ser Phe Tyr Leu Ile Asn Leu Ile Leu Ala Val
385 390 395 400
Val Ala Met Ala Tyr Glu Glu Gln Asn Gln Ala Asn Ile Glu Glu Ala
405 410 415
Lys Gln Lys Glu Leu Glu Phe Gln Gln Met Leu Asp Arg Leu Lys Lys
420 425 430
Glu Gln Glu Glu Ala Glu Ala Ile Ala Ala Ala Ala Ala Glu Tyr Thr
435 440 445
Ser Ile Arg Arg Ser Arg Ile Met Gly Leu Ser Glu Ser Ser Ser Glu
450 455 460
Thr Ser Lys Leu Ser Ser Lys Ser Ala Lys Glu Arg Arg Asn Arg Arg
465 470 475 480
Lys Lys Lys Asn Gln Lys Lys Leu Ser Ser Gly Glu Glu Lys Gly Asp
485 490 495
Ala Glu Lys Leu Ser Lys Ser Glu Ser Glu Asp Ser Ile Arg Arg Lys
500 505 510
Ser Phe His Leu Gly Val Glu Gly His Arg Arg Ala His Glu Lys Arg
515 520 525
Leu Ser Thr Pro Asn Gln Ser Pro Leu Ser Ile Arg Gly Ser Leu Phe
530 535 540
Ser Ala Arg Arg Ser Ser Arg Thr Ser Leu Phe Ser Phe Lys Gly Arg
545 550 555 560
Gly Arg Asp Ile Gly Ser Glu Thr Glu Phe Ala Asp Asp Glu His Ser
565 570 575
Ile Phe Gly Asp Asn Glu Ser Arg Arg Gly Ser Leu Phe Val Pro His
580 585 590
Arg Pro Gln Glu Arg Arg Ser Ser Asn Ile Ser Gln Ala Ser Arg Ser
595 600 605
Pro Pro Met Leu Pro Val Asn Gly Lys Met His Ser Ala Val Asp Cys
610 615 620
Asn Gly Val Val Ser Leu Val Asp Gly Arg Ser Ala Leu Met Leu Pro
625 630 635 640
Asn Gly Gln Leu Leu Pro Glu Val Ile Ile Asp Lys Ala Thr Ser Asp
645 650 655
Asp Ser Gly Thr Thr Asn Gln Ile His Lys Lys Arg Arg Cys Ser Ser
660 665 670
Tyr Leu Leu Ser Glu Asp Met Leu Asn Asp Pro Asn Leu Arg Gln Arg
675 680 685
Ala Met Ser Arg Ala Ser Ile Leu Thr Asn Thr Val Glu Glu Leu Glu
690 695 700
Glu Ser Arg Gln Lys Cys Pro Pro Trp Trp Tyr Arg Phe Ala His Lys
705 710 715 720
Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu Phe
725 730 735
Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile Ile
740 745 750
Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro Met
755 760 765
Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe Ile
770 775 780
Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His Pro
785 790 795 800
Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile Val
805 810 815
Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met Ala
820 825 830
Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys Tyr
835 840 845
Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp Val
850 855 860
Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe Ser
865 870 875 880
Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe Val
885 890 895
Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His Asp
900 905 910
Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu Trp
915 920 925
Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp Cys
930 935 940
Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val Leu
945 950 955 960
Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp Val
965 970 975
Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val Ala
980 985 990
Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys Lys
995 1000 1005
Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
1010 1015 1020
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser
1025 1030 1035
Asn Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr
1040 1045 1050
Glu Lys Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser
1055 1060 1065
Pro Ser Val Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp
1070 1075 1080
Ile Glu Asn Leu Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp
1085 1090 1095
Gly Gly Ser Lys Glu Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys
1100 1105 1110
Ser Thr Val Asp Ile Ala Ile Ser Glu Glu Glu Glu Met Phe Tyr
1115 1120 1125
Gly Gly Glu Arg Ser Lys His Leu Lys Asn Gly Cys Arg Arg Gly
1130 1135 1140
Ser Ser Leu Gly Gln Ile Ser Gly Ala Ser Lys Lys Gly Lys Ile
1145 1150 1155
Trp Gln Asn Ile Arg Lys Thr Cys Cys Lys Ile Val Glu Asn Asn
1160 1165 1170
Trp Phe Lys Cys Phe Ile Gly Leu Val Thr Leu Leu Ser Thr Gly
1175 1180 1185
Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln Arg Lys Thr Ile
1190 1195 1200
Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr Tyr Ile Phe
1205 1210 1215
Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe Lys Ala
1220 1225 1230
Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val Ile
1235 1240 1245
Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg Glu Glu Leu Lys
1250 1255 1260
Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu Ser
1265 1270 1275
Gln Phe Glu Arg Met Lys Val Val Val Arg Ala Leu Ile Lys Thr
1280 1285 1290
Thr Leu Pro Thr Leu Asn Val Phe Leu Val Cys Leu Met Ile Trp
1295 1300 1305
Leu Ile Phe Ser Ile Met Gly Val Asp Leu Phe Ala Gly Arg Phe
1310 1315 1320
Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg Phe Pro Ser Ser
1325 1330 1335
Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu Leu Phe Asn Glu
1340 1345 1350
Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe Asp Asn Val Gly
1355 1360 1365
Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr Phe Asn Gly Trp
1370 1375 1380
Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val Ala Val Asn Ile
1385 1390 1395
Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr Cys Tyr Phe Ile
1400 1405 1410
Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu Ser Met Leu Ile
1415 1420 1425
Thr Val Ile Ile Asp Asn Phe Asn Lys His Lys Ile Lys Leu Gly
1430 1435 1440
Gly Ser Asn Ile Phe Ile Thr Val Lys Gln Arg Lys Gln Tyr Arg
1445 1450 1455
Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser Gln Arg Pro Val Pro
1460 1465 1470
Arg Pro Leu Asn Lys Leu Gln Gly Phe Ile Phe Asp Val Val Thr
1475 1480 1485
Ser Gln Ala Phe Asn Val Ile Val Met Val Leu Ile Cys Phe Gln
1490 1495 1500
Ala Ile Ala Met Met Ile Asp Thr Asp Val Gln Ser Leu Gln Met
1505 1510 1515
Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile Phe Val Met Leu Tyr
1520 1525 1530
Thr Met Glu Cys Ile Leu Lys Leu Ile Ala Phe Arg Cys Phe Tyr
1535 1540 1545
Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe Met Val Val Ile Phe
1550 1555 1560
Ser Ile Thr Gly Leu Cys Leu Pro Met Thr Val Gly Ser Tyr Leu
1565 1570 1575
Val Pro Pro Ser Leu Val Gln Leu Ile Leu Leu Ser Arg Ile Ile
1580 1585 1590
His Met Leu Arg Leu Gly Lys Gly Pro Lys Val Phe His Asn Leu
1595 1600 1605
Met Leu Pro Leu Met Leu Ser Leu Pro Ala Leu Leu Asn Ile Ile
1610 1615 1620
Leu Leu Ile Phe Leu Val Met Phe Ile Tyr Ala Val Phe Gly Met
1625 1630 1635
Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala Gly Ile Asn Asp Val
1640 1645 1650
Ser Asn Phe Glu Thr Phe Gly Asn Ser Met Leu Cys Leu Phe Gln
1655 1660 1665
Val Ala Ile Phe Ala Gly Trp Asp Gly Met Leu Asp Ala Ile Phe
1670 1675 1680
Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp Lys Ile Asn Pro Gly
1685 1690 1695
Thr Gln Val Arg Gly Asp Cys Gly Asn Pro Ser Val Gly Ile Phe
1700 1705 1710
Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp Leu Ile Ile Val Asn
1715 1720 1725
Met Tyr Ile Val Val Val Met Glu Phe Leu Asn Ile Ala Ser Lys
1730 1735 1740
Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp Phe Arg Lys Phe Phe
1745 1750 1755
Gln Val Trp Lys Arg Phe Asp Pro Asp Arg Thr Gln Tyr Ile Asp
1760 1765 1770
Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu Asp Pro Pro Leu
1775 1780 1785
Phe Met Ala Lys Pro Asn Lys Gly Gln Leu Ile Ala Leu Asp Leu
1790 1795 1800
Pro Met Ala Val Gly Asp Arg Ile His Cys Leu Asp Ile Leu Leu
1805 1810 1815
Ala Phe Thr Lys Arg Val Met Gly Gln Asp Val Arg Met Glu Lys
1820 1825 1830
Val Val Ser Glu Ile Glu Ser Gly Phe Leu Leu Ala Asn Pro Phe
1835 1840 1845
Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr Leu Lys Arg Lys Gln
1850 1855 1860
Glu Ala Val Ser Ala Thr Ile Ile Gln Arg Ala Tyr Lys Asn Tyr
1865 1870 1875
Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser Asp Ile His Met Ile
1880 1885 1890
Asp Gly Asp Arg Asp Val His Ala Thr Lys Glu Gly Ala Tyr Phe
1895 1900 1905
Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln Ser Gln Ile
1910 1915 1920
<210> 7
<211> 1701
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX-hNaV1.7_DII_嵌合体
<400> 7
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Tyr Trp Ile Lys Phe Lys Lys
485 490 495
Cys Ile Tyr Phe Ile Val Met Asp Pro Phe Val Asp Leu Ala Ile Thr
500 505 510
Ile Cys Ile Val Leu Asn Thr Leu Phe Met Ala Met Glu His His Pro
515 520 525
Met Thr Glu Glu Phe Lys Asn Val Leu Ala Ile Gly Asn Leu Val Phe
530 535 540
Thr Gly Ile Phe Ala Ala Glu Met Val Leu Lys Leu Ile Ala Met Asp
545 550 555 560
Pro Tyr Glu Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Leu Ile
565 570 575
Val Thr Leu Ser Leu Val Glu Leu Phe Leu Ala Asp Val Glu Gly Leu
580 585 590
Ser Val Leu Arg Ser Phe Arg Leu Leu Arg Val Phe Lys Leu Ala Lys
595 600 605
Ser Trp Pro Thr Leu Asn Met Leu Ile Lys Ile Ile Gly Asn Ser Val
610 615 620
Gly Ala Leu Gly Asn Leu Thr Leu Val Leu Ala Ile Ile Val Phe Ile
625 630 635 640
Phe Ala Val Val Gly Met Gln Leu Phe Gly Lys Ser Tyr Lys Glu Cys
645 650 655
Val Cys Lys Ile Asn Asp Asp Cys Thr Leu Pro Arg Trp His Met Asn
660 665 670
Asp Phe Phe His Ser Phe Leu Ile Val Phe Arg Val Leu Cys Gly Glu
675 680 685
Trp Ile Glu Thr Met Trp Asp Cys Met Glu Val Ala Gly Gln Ala Met
690 695 700
Cys Leu Ile Val Tyr Met Met Val Met Val Ile Gly Asn Leu Val Val
705 710 715 720
Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe Ser Ser Asp Asn
725 730 735
Leu Thr Ala Ile Glu Glu Asp Pro Asp Ala Asn Asn Leu Gln Ile Ala
740 745 750
Val Thr Arg Ile Lys Lys Gly Ile Asn Tyr Val Lys Gln Thr Leu Arg
755 760 765
Glu Phe Ile Leu Lys Ala Phe Ser Lys Lys Pro Lys Ile Ser Arg Glu
770 775 780
Ile Arg Gln Ala Glu Asp Leu Asn Thr Lys Lys Glu Asn Tyr Ile Ser
785 790 795 800
Asn His Thr Leu Ala Glu Met Ser Lys Gly His Asn Phe Leu Lys Glu
805 810 815
Lys Asp Lys Ile Ser Gly Phe Gly Ser Ser Val Asp Lys His Leu Met
820 825 830
Glu Asp Ser Asp Gly Gln Ser Phe Ile His Asn Pro Ser Leu Thr Val
835 840 845
Thr Val Pro Ile Ala Pro Gly Glu Ser Asp Leu Glu Asn Met Asn Ala
850 855 860
Glu Glu Leu Ser Ser Asp Ser Asp Ser Glu Tyr Ser Lys Val Arg Leu
865 870 875 880
Asn Arg Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Asn Pro Leu Pro
885 890 895
Gly Glu Gly Glu Glu Ala Glu Ala Glu Pro Met Asn Ser Asp Glu Pro
900 905 910
Glu Ala Cys Phe Thr Asp Gly Cys Val Trp Arg Phe Ser Cys Cys Gln
915 920 925
Val Asn Ile Glu Ser Gly Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys
930 935 940
Thr Cys Cys Lys Ile Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly
945 950 955 960
Leu Val Thr Leu Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr
965 970 975
Met Asp Gln Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met
980 985 990
Ile Phe Thr Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala
995 1000 1005
Tyr Gly Phe Lys Ala Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp
1010 1015 1020
Phe Val Val Val Ile Val Phe Cys Leu Ser Leu Ile Gly Lys Thr
1025 1030 1035
Arg Glu Glu Leu Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro
1040 1045 1050
Leu Arg Val Leu Ser Gln Phe Glu Arg Met Lys Val Val Val Arg
1055 1060 1065
Ala Leu Ile Lys Thr Thr Leu Pro Thr Leu Asn Val Phe Leu Val
1070 1075 1080
Cys Leu Met Ile Trp Leu Ile Phe Ser Ile Met Gly Val Asp Leu
1085 1090 1095
Phe Ala Gly Arg Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu
1100 1105 1110
Arg Phe Pro Ser Ser Glu Val Met Asn Lys Ser Arg Cys Glu Ser
1115 1120 1125
Leu Leu Phe Asn Glu Ser Met Leu Trp Glu Asn Ala Lys Met Asn
1130 1135 1140
Phe Asp Asn Val Gly Asn Gly Phe Leu Ser Leu Leu Gln Val Ala
1145 1150 1155
Thr Phe Asn Gly Trp Ile Thr Ile Met Asn Ser Ala Ile Asp Ser
1160 1165 1170
Val Ala Val Asn Ile Gln Pro His Phe Glu Val Asn Ile Tyr Met
1175 1180 1185
Tyr Cys Tyr Phe Ile Asn Phe Ile Ile Phe Gly Val Phe Leu Pro
1190 1195 1200
Leu Ser Met Leu Ile Thr Val Ile Ile Asp Asn Phe Asn Lys His
1205 1210 1215
Lys Ile Lys Leu Gly Gly Ser Asn Ile Phe Ile Thr Val Lys Gln
1220 1225 1230
Arg Lys Gln Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser
1235 1240 1245
Gln Arg Pro Val Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe Ile
1250 1255 1260
Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val Met Val
1265 1270 1275
Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp Val
1280 1285 1290
Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile
1295 1300 1305
Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile Ala
1310 1315 1320
Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe
1325 1330 1335
Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met Thr
1340 1345 1350
Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile Leu
1355 1360 1365
Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro Lys
1370 1375 1380
Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro Ala
1385 1390 1395
Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile Tyr
1400 1405 1410
Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala
1415 1420 1425
Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser Met
1430 1435 1440
Leu Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly Met
1445 1450 1455
Leu Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp
1460 1465 1470
Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn Pro
1475 1480 1485
Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp
1490 1495 1500
Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu Phe Leu
1505 1510 1515
Asn Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp
1520 1525 1530
Phe Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro Asp Arg
1535 1540 1545
Thr Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala
1550 1555 1560
Leu Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly Gln Leu
1565 1570 1575
Ile Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile His Cys
1580 1585 1590
Leu Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly Gln Asp
1595 1600 1605
Val Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly Phe Leu
1610 1615 1620
Leu Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr
1625 1630 1635
Leu Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile Gln Arg
1640 1645 1650
Ala Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser
1655 1660 1665
Asp Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala Thr Lys
1670 1675 1680
Glu Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln
1685 1690 1695
Ser Gln Ile
1700
<210> 8
<211> 1688
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX-hNaV1.7_DIII_嵌合体
<400> 8
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys Thr Cys Tyr
915 920 925
Lys Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile Val Leu Met Ile
930 935 940
Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Arg
945 950 955 960
Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala Tyr Gly Tyr
980 985 990
Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val
995 1000 1005
Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu Gly Tyr Ser
1010 1015 1020
Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg
1025 1030 1035
Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val
1040 1045 1050
Asn Ala Leu Ile Gly Ala Ile Pro Ser Ile Met Asn Val Leu Leu
1055 1060 1065
Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile Met Gly Val Asn
1070 1075 1080
Leu Phe Ala Gly Lys Phe Tyr Glu Cys Ile Asn Thr Thr Asp Gly
1085 1090 1095
Ser Arg Phe Pro Ala Ser Gln Val Pro Asn Arg Ser Glu Cys Phe
1100 1105 1110
Ala Leu Met Asn Val Ser Gln Asn Val Arg Trp Lys Asn Leu Lys
1115 1120 1125
Val Asn Phe Asp Asn Val Gly Leu Gly Tyr Leu Ser Leu Leu Gln
1130 1135 1140
Val Ala Thr Phe Lys Gly Trp Thr Ile Ile Met Tyr Ala Ala Val
1145 1150 1155
Asp Ser Val Asn Val Asp Lys Gln Pro Lys Tyr Glu Tyr Ser Leu
1160 1165 1170
Tyr Met Tyr Ile Tyr Phe Val Val Phe Ile Ile Phe Gly Ser Phe
1175 1180 1185
Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe Asn
1190 1195 1200
Gln Gln Lys Lys Lys Leu Gly Gly Gln Asp Ile Phe Met Thr Glu
1205 1210 1215
Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys
1220 1225 1230
Lys Pro Gln Lys Pro Ile Pro Arg Pro Gly Asn Lys Leu Gln Gly
1235 1240 1245
Phe Ile Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val
1250 1255 1260
Met Val Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr
1265 1270 1275
Asp Val Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn
1280 1285 1290
Ser Ile Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu
1295 1300 1305
Ile Ala Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe
1310 1315 1320
Asp Phe Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro
1325 1330 1335
Met Thr Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu
1340 1345 1350
Ile Leu Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly
1355 1360 1365
Pro Lys Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu
1370 1375 1380
Pro Ala Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe
1385 1390 1395
Ile Tyr Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys
1400 1405 1410
Glu Ala Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn
1415 1420 1425
Ser Met Leu Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp
1430 1435 1440
Gly Met Leu Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp
1445 1450 1455
Pro Asp Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly
1460 1465 1470
Asn Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile
1475 1480 1485
Ser Trp Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu
1490 1495 1500
Phe Leu Asn Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu
1505 1510 1515
Asp Asp Phe Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro
1520 1525 1530
Asp Arg Thr Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala
1535 1540 1545
Ala Ala Leu Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly
1550 1555 1560
Gln Leu Ile Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile
1565 1570 1575
His Cys Leu Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly
1580 1585 1590
Gln Asp Val Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly
1595 1600 1605
Phe Leu Leu Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr
1610 1615 1620
Thr Thr Leu Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile
1625 1630 1635
Gln Arg Ala Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn
1640 1645 1650
Thr Ser Asp Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala
1655 1660 1665
Thr Lys Glu Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro
1670 1675 1680
Ile Gln Ser Gln Ile
1685
<210> 9
<211> 1724
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX-hNaV1.7_DIV-DIII-DIV接头_嵌合体
<400> 9
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys Thr Cys Cys
915 920 925
Lys Ile Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly Leu Val Thr
930 935 940
Leu Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln
945 950 955 960
Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe
980 985 990
Lys Ala Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val
995 1000 1005
Ile Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg Glu Glu Leu
1010 1015 1020
Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu
1025 1030 1035
Ser Gln Phe Glu Arg Met Lys Val Val Val Arg Ala Leu Ile Lys
1040 1045 1050
Thr Thr Leu Pro Thr Leu Asn Val Phe Leu Val Cys Leu Met Ile
1055 1060 1065
Trp Leu Ile Phe Ser Ile Met Gly Val Asp Leu Phe Ala Gly Arg
1070 1075 1080
Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg Phe Pro Ser
1085 1090 1095
Ser Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu Leu Phe Asn
1100 1105 1110
Glu Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe Asp Asn Val
1115 1120 1125
Gly Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr Phe Asn Gly
1130 1135 1140
Trp Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val Ala Val Asn
1145 1150 1155
Ile Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr Cys Tyr Phe
1160 1165 1170
Ile Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu Ser Met Leu
1175 1180 1185
Ile Thr Val Ile Ile Asp Asn Phe Asn Gln Gln Lys Lys Lys Leu
1190 1195 1200
Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr
1205 1210 1215
Asn Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile
1220 1225 1230
Pro Arg Pro Gly Asn Lys Ile Gln Gly Cys Ile Phe Asp Leu Val
1235 1240 1245
Thr Asn Gln Ala Phe Asp Ile Ser Ile Met Val Leu Ile Cys Leu
1250 1255 1260
Asn Met Val Thr Met Met Val Glu Lys Glu Gly Gln Ser Gln His
1265 1270 1275
Met Thr Glu Val Leu Tyr Trp Ile Asn Val Val Phe Ile Ile Leu
1280 1285 1290
Phe Thr Gly Glu Cys Val Leu Lys Leu Ile Ser Leu Arg His Tyr
1295 1300 1305
Tyr Phe Thr Val Gly Trp Asn Ile Phe Asp Phe Val Val Val Ile
1310 1315 1320
Ile Ser Ile Val Gly Met Phe Leu Ala Asp Leu Ile Glu Thr Tyr
1325 1330 1335
Phe Val Ser Pro Thr Leu Phe Arg Val Ile Arg Leu Ala Arg Ile
1340 1345 1350
Gly Arg Ile Leu Arg Leu Val Lys Gly Ala Lys Gly Ile Arg Thr
1355 1360 1365
Leu Leu Phe Ala Leu Met Met Ser Leu Pro Ala Leu Phe Asn Ile
1370 1375 1380
Gly Leu Leu Leu Phe Leu Val Met Phe Ile Tyr Ala Ile Phe Gly
1385 1390 1395
Met Ser Asn Phe Ala Tyr Val Lys Lys Glu Asp Gly Ile Asn Asp
1400 1405 1410
Met Phe Asn Phe Glu Thr Phe Gly Asn Ser Met Ile Cys Leu Phe
1415 1420 1425
Gln Ile Thr Thr Ser Ala Gly Trp Asp Gly Leu Leu Ala Pro Ile
1430 1435 1440
Leu Asn Ser Lys Pro Pro Asp Cys Asp Pro Lys Lys Val His Pro
1445 1450 1455
Gly Ser Ser Val Glu Gly Asp Cys Gly Asn Pro Ser Val Gly Ile
1460 1465 1470
Phe Tyr Phe Val Ser Tyr Ile Ile Ile Ser Phe Leu Val Val Val
1475 1480 1485
Asn Met Tyr Ile Ala Val Ile Leu Glu Asn Phe Ser Val Ala Thr
1490 1495 1500
Glu Glu Ser Thr Glu Pro Leu Ser Glu Asp Asp Phe Glu Met Phe
1505 1510 1515
Tyr Glu Val Trp Glu Lys Phe Asp Pro Asp Ala Thr Gln Phe Ile
1520 1525 1530
Glu Phe Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu Asp Pro Pro
1535 1540 1545
Leu Leu Ile Ala Lys Pro Asn Lys Val Gln Leu Ile Ala Met Asp
1550 1555 1560
Leu Pro Met Val Ser Gly Asp Arg Ile His Cys Leu Asp Ile Leu
1565 1570 1575
Phe Ala Phe Thr Lys Arg Val Leu Gly Glu Ser Gly Glu Met Asp
1580 1585 1590
Ser Leu Arg Ser Gln Met Glu Glu Arg Phe Met Ser Ala Asn Pro
1595 1600 1605
Ser Lys Val Ser Tyr Glu Pro Ile Thr Thr Thr Leu Lys Arg Lys
1610 1615 1620
Gln Glu Asp Val Ser Ala Thr Val Ile Gln Arg Ala Tyr Arg Arg
1625 1630 1635
Tyr Arg Leu Arg Gln Asn Val Lys Asn Ile Ser Ser Ile Tyr Ile
1640 1645 1650
Lys Asp Gly Asp Arg Asp Asp Asp Leu Leu Asn Lys Lys Asp Met
1655 1660 1665
Ala Phe Asp Asn Val Asn Glu Asn Ser Ser Pro Glu Lys Thr Asp
1670 1675 1680
Ala Thr Ser Ser Thr Thr Ser Pro Pro Ser Tyr Asp Ser Val Thr
1685 1690 1695
Lys Pro Asp Lys Glu Lys Tyr Glu Gln Asp Arg Thr Glu Lys Glu
1700 1705 1710
Asp Lys Gly Lys Asp Ser Lys Glu Ser Lys Lys
1715 1720
<210> 10
<211> 1688
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_DIII_嵌合体_SF_嵌合体
<400> 10
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Ser Trp Ala Phe Leu Ala Leu Phe Arg Leu Met Thr Gln
340 345 350
Asp Tyr Trp Glu Asn Leu Tyr Gln Gln Thr Leu Arg Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Ile Val Phe Arg Val Leu Cys Gly Glu
675 680 685
Trp Ile Glu Thr Met Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys Thr Cys Tyr
915 920 925
Lys Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile Val Leu Met Ile
930 935 940
Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Arg
945 950 955 960
Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala Tyr Gly Tyr
980 985 990
Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val
995 1000 1005
Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu Gly Tyr Ser
1010 1015 1020
Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg
1025 1030 1035
Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val
1040 1045 1050
Asn Ala Leu Ile Gly Ala Ile Pro Ser Ile Met Asn Val Leu Leu
1055 1060 1065
Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile Met Gly Val Asn
1070 1075 1080
Leu Phe Ala Gly Lys Phe Tyr Glu Cys Ile Asn Thr Thr Asp Gly
1085 1090 1095
Ser Arg Phe Pro Ala Ser Gln Val Pro Asn Arg Ser Glu Cys Phe
1100 1105 1110
Ala Leu Met Asn Val Ser Gln Asn Val Arg Trp Lys Asn Leu Lys
1115 1120 1125
Val Asn Phe Asp Asn Val Gly Leu Gly Tyr Leu Ser Leu Leu Gln
1130 1135 1140
Val Ala Thr Phe Lys Gly Trp Thr Ile Ile Met Tyr Ala Ala Val
1145 1150 1155
Asp Ser Val Asn Val Asp Lys Gln Pro Lys Tyr Glu Tyr Ser Leu
1160 1165 1170
Tyr Met Tyr Ile Tyr Phe Val Val Phe Ile Ile Phe Gly Ser Phe
1175 1180 1185
Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe Asn
1190 1195 1200
Gln Gln Lys Lys Lys Leu Gly Gly Gln Asp Ile Phe Met Thr Glu
1205 1210 1215
Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys
1220 1225 1230
Lys Pro Gln Lys Pro Ile Pro Arg Pro Gly Asn Lys Leu Gln Gly
1235 1240 1245
Phe Ile Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val
1250 1255 1260
Met Val Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr
1265 1270 1275
Asp Val Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn
1280 1285 1290
Ser Ile Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu
1295 1300 1305
Ile Ala Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe
1310 1315 1320
Asp Phe Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro
1325 1330 1335
Met Thr Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu
1340 1345 1350
Ile Leu Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly
1355 1360 1365
Pro Lys Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu
1370 1375 1380
Pro Ala Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe
1385 1390 1395
Ile Tyr Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys
1400 1405 1410
Glu Ala Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn
1415 1420 1425
Ser Met Ile Cys Leu Phe Gln Ile Thr Thr Ser Ala Gly Trp Asp
1430 1435 1440
Gly Leu Leu Ala Pro Ile Leu Asn Ser Lys Trp Ser Asp Cys Asp
1445 1450 1455
Pro Asp Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly
1460 1465 1470
Asn Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile
1475 1480 1485
Ser Trp Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu
1490 1495 1500
Phe Leu Asn Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu
1505 1510 1515
Asp Asp Phe Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro
1520 1525 1530
Asp Arg Thr Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala
1535 1540 1545
Ala Ala Leu Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly
1550 1555 1560
Gln Leu Ile Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile
1565 1570 1575
His Cys Leu Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly
1580 1585 1590
Gln Asp Val Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly
1595 1600 1605
Phe Leu Leu Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr
1610 1615 1620
Thr Thr Leu Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile
1625 1630 1635
Gln Arg Ala Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn
1640 1645 1650
Thr Ser Asp Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala
1655 1660 1665
Thr Lys Glu Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro
1670 1675 1680
Ile Gln Ser Gln Ile
1685
<210> 11
<211> 1688
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_D3-D4连接_嵌合体
<400> 11
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys Thr Cys Tyr
915 920 925
Lys Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile Val Leu Met Ile
930 935 940
Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Arg
945 950 955 960
Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala Tyr Gly Tyr
980 985 990
Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val
995 1000 1005
Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu Gly Tyr Ser
1010 1015 1020
Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg
1025 1030 1035
Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val
1040 1045 1050
Asn Ala Leu Ile Gly Ala Ile Pro Ser Ile Met Asn Val Leu Leu
1055 1060 1065
Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile Met Gly Val Asn
1070 1075 1080
Leu Phe Ala Gly Lys Phe Tyr Glu Cys Ile Asn Thr Thr Asp Gly
1085 1090 1095
Ser Arg Phe Pro Ala Ser Gln Val Pro Asn Arg Ser Glu Cys Phe
1100 1105 1110
Ala Leu Met Asn Val Ser Gln Asn Val Arg Trp Lys Asn Leu Lys
1115 1120 1125
Val Asn Phe Asp Asn Val Gly Leu Gly Tyr Leu Ser Leu Leu Gln
1130 1135 1140
Val Ala Thr Phe Lys Gly Trp Thr Ile Ile Met Tyr Ala Ala Val
1145 1150 1155
Asp Ser Val Asn Val Asp Lys Gln Pro Lys Tyr Glu Tyr Ser Leu
1160 1165 1170
Tyr Met Tyr Ile Tyr Phe Val Val Phe Ile Ile Phe Gly Ser Phe
1175 1180 1185
Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe Asn
1190 1195 1200
Gln Gln Lys Lys Lys Leu Gly Gly Ser Asn Ile Phe Ile Thr Val
1205 1210 1215
Lys Gln Arg Lys Gln Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu
1220 1225 1230
Asp Ser Gln Arg Pro Val Pro Arg Pro Leu Asn Lys Leu Gln Gly
1235 1240 1245
Phe Ile Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val
1250 1255 1260
Met Val Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr
1265 1270 1275
Asp Val Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn
1280 1285 1290
Ser Ile Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu
1295 1300 1305
Ile Ala Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe
1310 1315 1320
Asp Phe Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro
1325 1330 1335
Met Thr Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu
1340 1345 1350
Ile Leu Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly
1355 1360 1365
Pro Lys Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu
1370 1375 1380
Pro Ala Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe
1385 1390 1395
Ile Tyr Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys
1400 1405 1410
Glu Ala Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn
1415 1420 1425
Ser Met Leu Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp
1430 1435 1440
Gly Met Leu Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp
1445 1450 1455
Pro Asp Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly
1460 1465 1470
Asn Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile
1475 1480 1485
Ser Trp Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu
1490 1495 1500
Phe Leu Asn Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu
1505 1510 1515
Asp Asp Phe Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro
1520 1525 1530
Asp Arg Thr Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala
1535 1540 1545
Ala Ala Leu Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly
1550 1555 1560
Gln Leu Ile Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile
1565 1570 1575
His Cys Leu Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly
1580 1585 1590
Gln Asp Val Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly
1595 1600 1605
Phe Leu Leu Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr
1610 1615 1620
Thr Thr Leu Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile
1625 1630 1635
Gln Arg Ala Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn
1640 1645 1650
Thr Ser Asp Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala
1655 1660 1665
Thr Lys Glu Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro
1670 1675 1680
Ile Gln Ser Gln Ile
1685
<210> 12
<211> 1687
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_D3-D4连接-PM_嵌合体
<400> 12
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys Thr Cys Tyr
915 920 925
Lys Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile Val Leu Met Ile
930 935 940
Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Arg
945 950 955 960
Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala Tyr Gly Tyr
980 985 990
Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val
995 1000 1005
Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu Gly Tyr Ser
1010 1015 1020
Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg
1025 1030 1035
Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val
1040 1045 1050
Asn Ala Leu Ile Gly Ala Ile Leu Pro Thr Leu Asn Val Phe Leu
1055 1060 1065
Val Cys Leu Met Ile Trp Leu Ile Phe Ser Ile Met Gly Val Asp
1070 1075 1080
Leu Phe Ala Gly Arg Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly
1085 1090 1095
Glu Arg Phe Pro Ser Ser Glu Val Met Asn Lys Ser Arg Cys Glu
1100 1105 1110
Ser Leu Leu Phe Asn Glu Ser Met Leu Trp Glu Asn Ala Lys Met
1115 1120 1125
Asn Phe Asp Asn Val Gly Asn Gly Phe Leu Ser Leu Leu Gln Val
1130 1135 1140
Ala Thr Phe Asn Gly Trp Ile Thr Ile Met Asn Ser Ala Ile Asp
1145 1150 1155
Ser Val Ala Val Asn Ile Gln Pro His Phe Glu Val Asn Ile Tyr
1160 1165 1170
Met Tyr Cys Tyr Phe Ile Asn Phe Ile Ile Phe Gly Val Phe Leu
1175 1180 1185
Pro Leu Ser Met Leu Ile Thr Val Ile Ile Asp Asn Phe Asn Lys
1190 1195 1200
His Lys Ile Lys Leu Gly Gly Ser Asn Ile Phe Ile Thr Val Lys
1205 1210 1215
Gln Arg Lys Gln Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp
1220 1225 1230
Ser Gln Arg Pro Val Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe
1235 1240 1245
Ile Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val Met
1250 1255 1260
Val Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp
1265 1270 1275
Val Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser
1280 1285 1290
Ile Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile
1295 1300 1305
Ala Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp
1310 1315 1320
Phe Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met
1325 1330 1335
Thr Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile
1340 1345 1350
Leu Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro
1355 1360 1365
Lys Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro
1370 1375 1380
Ala Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile
1385 1390 1395
Tyr Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu
1400 1405 1410
Ala Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser
1415 1420 1425
Met Leu Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly
1430 1435 1440
Met Leu Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro
1445 1450 1455
Asp Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn
1460 1465 1470
Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser
1475 1480 1485
Trp Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu Phe
1490 1495 1500
Leu Asn Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp
1505 1510 1515
Asp Phe Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro Asp
1520 1525 1530
Arg Thr Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala
1535 1540 1545
Ala Leu Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly Gln
1550 1555 1560
Leu Ile Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile His
1565 1570 1575
Cys Leu Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly Gln
1580 1585 1590
Asp Val Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly Phe
1595 1600 1605
Leu Leu Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr
1610 1615 1620
Thr Leu Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile Gln
1625 1630 1635
Arg Ala Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr
1640 1645 1650
Ser Asp Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala Thr
1655 1660 1665
Lys Glu Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile
1670 1675 1680
Gln Ser Gln Ile
1685
<210> 13
<211> 1687
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_D3-D4连接-PM-S4S5连接_嵌合体
<400> 13
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys Thr Cys Tyr
915 920 925
Lys Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile Val Leu Met Ile
930 935 940
Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Arg
945 950 955 960
Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala Tyr Gly Tyr
980 985 990
Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val
995 1000 1005
Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu Gly Tyr Ser
1010 1015 1020
Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg
1025 1030 1035
Pro Leu Arg Ala Leu Ser Arg Phe Glu Arg Met Lys Val Val Val
1040 1045 1050
Arg Ala Leu Ile Lys Thr Thr Leu Pro Thr Leu Asn Val Phe Leu
1055 1060 1065
Val Cys Leu Met Ile Trp Leu Ile Phe Ser Ile Met Gly Val Asp
1070 1075 1080
Leu Phe Ala Gly Arg Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly
1085 1090 1095
Glu Arg Phe Pro Ser Ser Glu Val Met Asn Lys Ser Arg Cys Glu
1100 1105 1110
Ser Leu Leu Phe Asn Glu Ser Met Leu Trp Glu Asn Ala Lys Met
1115 1120 1125
Asn Phe Asp Asn Val Gly Asn Gly Phe Leu Ser Leu Leu Gln Val
1130 1135 1140
Ala Thr Phe Asn Gly Trp Ile Thr Ile Met Asn Ser Ala Ile Asp
1145 1150 1155
Ser Val Ala Val Asn Ile Gln Pro His Phe Glu Val Asn Ile Tyr
1160 1165 1170
Met Tyr Cys Tyr Phe Ile Asn Phe Ile Ile Phe Gly Val Phe Leu
1175 1180 1185
Pro Leu Ser Met Leu Ile Thr Val Ile Ile Asp Asn Phe Asn Lys
1190 1195 1200
His Lys Ile Lys Leu Gly Gly Ser Asn Ile Phe Ile Thr Val Lys
1205 1210 1215
Gln Arg Lys Gln Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp
1220 1225 1230
Ser Gln Arg Pro Val Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe
1235 1240 1245
Ile Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val Met
1250 1255 1260
Val Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp
1265 1270 1275
Val Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser
1280 1285 1290
Ile Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile
1295 1300 1305
Ala Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp
1310 1315 1320
Phe Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met
1325 1330 1335
Thr Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile
1340 1345 1350
Leu Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro
1355 1360 1365
Lys Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro
1370 1375 1380
Ala Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile
1385 1390 1395
Tyr Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu
1400 1405 1410
Ala Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser
1415 1420 1425
Met Leu Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly
1430 1435 1440
Met Leu Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro
1445 1450 1455
Asp Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn
1460 1465 1470
Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser
1475 1480 1485
Trp Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu Phe
1490 1495 1500
Leu Asn Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp
1505 1510 1515
Asp Phe Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro Asp
1520 1525 1530
Arg Thr Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala
1535 1540 1545
Ala Leu Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly Gln
1550 1555 1560
Leu Ile Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile His
1565 1570 1575
Cys Leu Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly Gln
1580 1585 1590
Asp Val Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly Phe
1595 1600 1605
Leu Leu Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr
1610 1615 1620
Thr Leu Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile Gln
1625 1630 1635
Arg Ala Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr
1640 1645 1650
Ser Asp Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala Thr
1655 1660 1665
Lys Glu Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile
1670 1675 1680
Gln Ser Gln Ile
1685
<210> 14
<211> 1687
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_VSD3_嵌合体
<400> 14
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys Thr Cys Tyr
915 920 925
Lys Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile Val Leu Met Ile
930 935 940
Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Arg
945 950 955 960
Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala Tyr Gly Tyr
980 985 990
Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val
995 1000 1005
Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu Gly Tyr Ser
1010 1015 1020
Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg
1025 1030 1035
Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Lys Val Val Val
1040 1045 1050
Arg Ala Leu Ile Lys Thr Thr Leu Pro Thr Leu Asn Val Phe Leu
1055 1060 1065
Val Cys Leu Met Ile Trp Leu Ile Phe Ser Ile Met Gly Val Asp
1070 1075 1080
Leu Phe Ala Gly Arg Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly
1085 1090 1095
Glu Arg Phe Pro Ser Ser Glu Val Met Asn Lys Ser Arg Cys Glu
1100 1105 1110
Ser Leu Leu Phe Asn Glu Ser Met Leu Trp Glu Asn Ala Lys Met
1115 1120 1125
Asn Phe Asp Asn Val Gly Asn Gly Phe Leu Ser Leu Leu Gln Val
1130 1135 1140
Ala Thr Phe Asn Gly Trp Ile Thr Ile Met Asn Ser Ala Ile Asp
1145 1150 1155
Ser Val Ala Val Asn Ile Gln Pro His Phe Glu Val Asn Ile Tyr
1160 1165 1170
Met Tyr Cys Tyr Phe Ile Asn Phe Ile Ile Phe Gly Val Phe Leu
1175 1180 1185
Pro Leu Ser Met Leu Ile Thr Val Ile Ile Asp Asn Phe Asn Lys
1190 1195 1200
His Lys Ile Lys Leu Gly Gly Ser Asn Ile Phe Ile Thr Val Lys
1205 1210 1215
Gln Arg Lys Gln Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp
1220 1225 1230
Ser Gln Arg Pro Val Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe
1235 1240 1245
Ile Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val Met
1250 1255 1260
Val Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp
1265 1270 1275
Val Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser
1280 1285 1290
Ile Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile
1295 1300 1305
Ala Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp
1310 1315 1320
Phe Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met
1325 1330 1335
Thr Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile
1340 1345 1350
Leu Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro
1355 1360 1365
Lys Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro
1370 1375 1380
Ala Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile
1385 1390 1395
Tyr Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu
1400 1405 1410
Ala Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser
1415 1420 1425
Met Leu Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly
1430 1435 1440
Met Leu Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro
1445 1450 1455
Asp Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn
1460 1465 1470
Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser
1475 1480 1485
Trp Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu Phe
1490 1495 1500
Leu Asn Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp
1505 1510 1515
Asp Phe Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro Asp
1520 1525 1530
Arg Thr Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala
1535 1540 1545
Ala Leu Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly Gln
1550 1555 1560
Leu Ile Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile His
1565 1570 1575
Cys Leu Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly Gln
1580 1585 1590
Asp Val Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly Phe
1595 1600 1605
Leu Leu Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr
1610 1615 1620
Thr Leu Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile Gln
1625 1630 1635
Arg Ala Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr
1640 1645 1650
Ser Asp Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala Thr
1655 1660 1665
Lys Glu Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile
1670 1675 1680
Gln Ser Gln Ile
1685
<210> 15
<211> 1687
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_VSD3-S4S5连接_嵌合体
<400> 15
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys Thr Cys Tyr
915 920 925
Lys Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile Val Leu Met Ile
930 935 940
Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Arg
945 950 955 960
Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala Tyr Gly Tyr
980 985 990
Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val
995 1000 1005
Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu Gly Tyr Ser
1010 1015 1020
Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg
1025 1030 1035
Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val
1040 1045 1050
Asn Ala Leu Ile Gly Ala Ile Pro Ser Thr Leu Asn Val Phe Leu
1055 1060 1065
Val Cys Leu Met Ile Trp Leu Ile Phe Ser Ile Met Gly Val Asp
1070 1075 1080
Leu Phe Ala Gly Arg Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly
1085 1090 1095
Glu Arg Phe Pro Ser Ser Glu Val Met Asn Lys Ser Arg Cys Glu
1100 1105 1110
Ser Leu Leu Phe Asn Glu Ser Met Leu Trp Glu Asn Ala Lys Met
1115 1120 1125
Asn Phe Asp Asn Val Gly Asn Gly Phe Leu Ser Leu Leu Gln Val
1130 1135 1140
Ala Thr Phe Asn Gly Trp Ile Thr Ile Met Asn Ser Ala Ile Asp
1145 1150 1155
Ser Val Ala Val Asn Ile Gln Pro His Phe Glu Val Asn Ile Tyr
1160 1165 1170
Met Tyr Cys Tyr Phe Ile Asn Phe Ile Ile Phe Gly Val Phe Leu
1175 1180 1185
Pro Leu Ser Met Leu Ile Thr Val Ile Ile Asp Asn Phe Asn Lys
1190 1195 1200
His Lys Ile Lys Leu Gly Gly Ser Asn Ile Phe Ile Thr Val Lys
1205 1210 1215
Gln Arg Lys Gln Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp
1220 1225 1230
Ser Gln Arg Pro Val Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe
1235 1240 1245
Ile Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val Met
1250 1255 1260
Val Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp
1265 1270 1275
Val Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser
1280 1285 1290
Ile Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile
1295 1300 1305
Ala Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp
1310 1315 1320
Phe Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met
1325 1330 1335
Thr Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile
1340 1345 1350
Leu Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro
1355 1360 1365
Lys Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro
1370 1375 1380
Ala Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile
1385 1390 1395
Tyr Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu
1400 1405 1410
Ala Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser
1415 1420 1425
Met Leu Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly
1430 1435 1440
Met Leu Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro
1445 1450 1455
Asp Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn
1460 1465 1470
Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser
1475 1480 1485
Trp Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu Phe
1490 1495 1500
Leu Asn Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp
1505 1510 1515
Asp Phe Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro Asp
1520 1525 1530
Arg Thr Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala
1535 1540 1545
Ala Leu Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly Gln
1550 1555 1560
Leu Ile Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile His
1565 1570 1575
Cys Leu Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly Gln
1580 1585 1590
Asp Val Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly Phe
1595 1600 1605
Leu Leu Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr
1610 1615 1620
Thr Leu Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile Gln
1625 1630 1635
Arg Ala Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr
1640 1645 1650
Ser Asp Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala Thr
1655 1660 1665
Lys Glu Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile
1670 1675 1680
Gln Ser Gln Ile
1685
<210> 16
<211> 1688
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_VSD3-S4S5连接-PM_嵌合体
<400> 16
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys Thr Cys Tyr
915 920 925
Lys Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile Val Leu Met Ile
930 935 940
Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Arg
945 950 955 960
Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala Tyr Gly Tyr
980 985 990
Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val
995 1000 1005
Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu Gly Tyr Ser
1010 1015 1020
Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg
1025 1030 1035
Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val
1040 1045 1050
Asn Ala Leu Ile Gly Ala Ile Pro Ser Ile Met Asn Val Leu Leu
1055 1060 1065
Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile Met Gly Val Asn
1070 1075 1080
Leu Phe Ala Gly Lys Phe Tyr Glu Cys Ile Asn Thr Thr Asp Gly
1085 1090 1095
Ser Arg Phe Pro Ala Ser Gln Val Pro Asn Arg Ser Glu Cys Phe
1100 1105 1110
Ala Leu Met Asn Val Ser Gln Asn Val Arg Trp Lys Asn Leu Lys
1115 1120 1125
Val Asn Phe Asp Asn Val Gly Leu Gly Tyr Leu Ser Leu Leu Gln
1130 1135 1140
Val Ala Thr Phe Lys Gly Trp Thr Ile Ile Met Tyr Ala Ala Val
1145 1150 1155
Asp Ser Val Asn Val Asp Lys Gln Pro Lys Tyr Glu Tyr Ser Leu
1160 1165 1170
Tyr Met Tyr Ile Tyr Phe Val Val Phe Ile Ile Phe Gly Ser Phe
1175 1180 1185
Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe Asn
1190 1195 1200
Gln Gln Lys Lys Lys Leu Gly Gly Gln Asp Ile Phe Met Thr Val
1205 1210 1215
Lys Gln Arg Lys Gln Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu
1220 1225 1230
Asp Ser Gln Arg Pro Val Pro Arg Pro Leu Asn Lys Leu Gln Gly
1235 1240 1245
Phe Ile Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val
1250 1255 1260
Met Val Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr
1265 1270 1275
Asp Val Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn
1280 1285 1290
Ser Ile Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu
1295 1300 1305
Ile Ala Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe
1310 1315 1320
Asp Phe Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro
1325 1330 1335
Met Thr Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu
1340 1345 1350
Ile Leu Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly
1355 1360 1365
Pro Lys Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu
1370 1375 1380
Pro Ala Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe
1385 1390 1395
Ile Tyr Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys
1400 1405 1410
Glu Ala Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn
1415 1420 1425
Ser Met Leu Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp
1430 1435 1440
Gly Met Leu Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp
1445 1450 1455
Pro Asp Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly
1460 1465 1470
Asn Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile
1475 1480 1485
Ser Trp Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu
1490 1495 1500
Phe Leu Asn Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu
1505 1510 1515
Asp Asp Phe Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro
1520 1525 1530
Asp Arg Thr Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala
1535 1540 1545
Ala Ala Leu Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly
1550 1555 1560
Gln Leu Ile Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile
1565 1570 1575
His Cys Leu Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly
1580 1585 1590
Gln Asp Val Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly
1595 1600 1605
Phe Leu Leu Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr
1610 1615 1620
Thr Thr Leu Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile
1625 1630 1635
Gln Arg Ala Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn
1640 1645 1650
Thr Ser Asp Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala
1655 1660 1665
Thr Lys Glu Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro
1670 1675 1680
Ile Gln Ser Gln Ile
1685
<210> 17
<211> 1687
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_DIII_VSD3-S3-S4_嵌合体
<400> 17
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys Thr Cys Cys
915 920 925
Lys Ile Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly Leu Val Thr
930 935 940
Leu Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln
945 950 955 960
Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe
980 985 990
Lys Ala Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val
995 1000 1005
Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu Gly Tyr Ser
1010 1015 1020
Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg
1025 1030 1035
Pro Leu Arg Ala Leu Ser Arg Phe Glu Arg Met Lys Val Val Val
1040 1045 1050
Arg Ala Leu Ile Lys Thr Thr Leu Pro Thr Leu Asn Val Phe Leu
1055 1060 1065
Val Cys Leu Met Ile Trp Leu Ile Phe Ser Ile Met Gly Val Asp
1070 1075 1080
Leu Phe Ala Gly Arg Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly
1085 1090 1095
Glu Arg Phe Pro Ser Ser Glu Val Met Asn Lys Ser Arg Cys Glu
1100 1105 1110
Ser Leu Leu Phe Asn Glu Ser Met Leu Trp Glu Asn Ala Lys Met
1115 1120 1125
Asn Phe Asp Asn Val Gly Asn Gly Phe Leu Ser Leu Leu Gln Val
1130 1135 1140
Ala Thr Phe Asn Gly Trp Ile Thr Ile Met Asn Ser Ala Ile Asp
1145 1150 1155
Ser Val Ala Val Asn Ile Gln Pro His Phe Glu Val Asn Ile Tyr
1160 1165 1170
Met Tyr Cys Tyr Phe Ile Asn Phe Ile Ile Phe Gly Val Phe Leu
1175 1180 1185
Pro Leu Ser Met Leu Ile Thr Val Ile Ile Asp Asn Phe Asn Lys
1190 1195 1200
His Lys Ile Lys Leu Gly Gly Ser Asn Ile Phe Ile Thr Val Lys
1205 1210 1215
Gln Arg Lys Gln Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp
1220 1225 1230
Ser Gln Arg Pro Val Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe
1235 1240 1245
Ile Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val Met
1250 1255 1260
Val Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp
1265 1270 1275
Val Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser
1280 1285 1290
Ile Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile
1295 1300 1305
Ala Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp
1310 1315 1320
Phe Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met
1325 1330 1335
Thr Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile
1340 1345 1350
Leu Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro
1355 1360 1365
Lys Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro
1370 1375 1380
Ala Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile
1385 1390 1395
Tyr Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu
1400 1405 1410
Ala Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser
1415 1420 1425
Met Leu Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly
1430 1435 1440
Met Leu Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro
1445 1450 1455
Asp Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn
1460 1465 1470
Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser
1475 1480 1485
Trp Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu Phe
1490 1495 1500
Leu Asn Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp
1505 1510 1515
Asp Phe Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro Asp
1520 1525 1530
Arg Thr Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala
1535 1540 1545
Ala Leu Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly Gln
1550 1555 1560
Leu Ile Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile His
1565 1570 1575
Cys Leu Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly Gln
1580 1585 1590
Asp Val Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly Phe
1595 1600 1605
Leu Leu Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr
1610 1615 1620
Thr Leu Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile Gln
1625 1630 1635
Arg Ala Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr
1640 1645 1650
Ser Asp Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala Thr
1655 1660 1665
Lys Glu Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile
1670 1675 1680
Gln Ser Gln Ile
1685
<210> 18
<211> 1683
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_DIII_VSD3-S5-S6_嵌合体
<400> 18
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys Thr Cys Cys
915 920 925
Lys Ile Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly Leu Val Thr
930 935 940
Leu Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln
945 950 955 960
Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe
980 985 990
Lys Ala Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val
995 1000 1005
Ile Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg Glu Glu Leu
1010 1015 1020
Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu
1025 1030 1035
Ser Gln Phe Glu Gly Met Arg Val Val Val Asn Ala Leu Ile Gly
1040 1045 1050
Ala Ile Pro Ser Ile Met Asn Val Leu Leu Val Cys Leu Ile Phe
1055 1060 1065
Trp Leu Ile Phe Ser Ile Met Gly Val Asn Leu Phe Ala Gly Lys
1070 1075 1080
Phe Tyr Glu Cys Ile Asn Thr Thr Asp Gly Ser Arg Phe Pro Ala
1085 1090 1095
Ser Gln Val Pro Asn Arg Ser Glu Cys Phe Ala Leu Met Asn Val
1100 1105 1110
Ser Gln Asn Val Arg Trp Lys Asn Leu Lys Val Asn Phe Asp Asn
1115 1120 1125
Val Gly Leu Gly Tyr Leu Ser Leu Leu Gln Val Ala Thr Phe Lys
1130 1135 1140
Gly Trp Thr Ile Ile Met Tyr Ala Ala Val Asp Ser Val Asn Val
1145 1150 1155
Asp Lys Gln Pro Lys Tyr Glu Tyr Ser Leu Tyr Met Tyr Ile Tyr
1160 1165 1170
Phe Val Val Phe Ile Ile Phe Gly Ser Phe Phe Thr Leu Asn Leu
1175 1180 1185
Phe Ile Gly Val Ile Ile Asp Asn Phe Asn Gln Gln Lys Lys Lys
1190 1195 1200
Leu Gly Gly Ser Asn Ile Phe Ile Thr Val Lys Gln Arg Lys Gln
1205 1210 1215
Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser Gln Arg Pro
1220 1225 1230
Val Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe Ile Phe Asp Val
1235 1240 1245
Val Thr Ser Gln Ala Phe Asn Val Ile Val Met Val Leu Ile Cys
1250 1255 1260
Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp Val Gln Ser Leu
1265 1270 1275
Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile Phe Val Met
1280 1285 1290
Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile Ala Phe Arg Cys
1295 1300 1305
Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe Met Val Val
1310 1315 1320
Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met Thr Val Gly Ser
1325 1330 1335
Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile Leu Leu Ser Arg
1340 1345 1350
Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro Lys Val Phe His
1355 1360 1365
Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro Ala Leu Leu Asn
1370 1375 1380
Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile Tyr Ala Val Phe
1385 1390 1395
Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala Gly Ile Asn
1400 1405 1410
Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser Met Leu Cys Leu
1415 1420 1425
Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly Met Leu Asp Ala
1430 1435 1440
Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp Lys Ile Asn
1445 1450 1455
Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn Pro Ser Val Gly
1460 1465 1470
Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp Leu Ile Ile
1475 1480 1485
Val Asn Met Tyr Ile Val Val Val Met Glu Phe Leu Asn Ile Ala
1490 1495 1500
Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp Phe Arg Lys
1505 1510 1515
Phe Phe Gln Val Trp Lys Arg Phe Asp Pro Asp Arg Thr Gln Tyr
1520 1525 1530
Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu Asp Pro
1535 1540 1545
Pro Leu Phe Met Ala Lys Pro Asn Lys Gly Gln Leu Ile Ala Leu
1550 1555 1560
Asp Leu Pro Met Ala Val Gly Asp Arg Ile His Cys Leu Asp Ile
1565 1570 1575
Leu Leu Ala Phe Thr Lys Arg Val Met Gly Gln Asp Val Arg Met
1580 1585 1590
Glu Lys Val Val Ser Glu Ile Glu Ser Gly Phe Leu Leu Ala Asn
1595 1600 1605
Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr Leu Lys Arg
1610 1615 1620
Lys Gln Glu Ala Val Ser Ala Thr Ile Ile Gln Arg Ala Tyr Lys
1625 1630 1635
Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser Asp Ile His
1640 1645 1650
Met Ile Asp Gly Asp Arg Asp Val His Ala Thr Lys Glu Gly Ala
1655 1660 1665
Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln Ser Gln Ile
1670 1675 1680
<210> 19
<211> 1683
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_DIII_VSD3-S3-S4环_S4-S5连接_S5_嵌合体
<400> 19
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys Thr Cys Cys
915 920 925
Lys Ile Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly Leu Val Thr
930 935 940
Leu Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln
945 950 955 960
Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe
980 985 990
Lys Ala Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val
995 1000 1005
Ile Val Phe Cys Leu Ser Leu Ile Gly Gly Tyr Ser Asp Leu Gly
1010 1015 1020
Leu Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val
1025 1030 1035
Leu Ser Gln Phe Glu Gly Met Arg Val Val Val Asn Ala Leu Ile
1040 1045 1050
Gly Ala Ile Pro Ser Ile Met Asn Val Leu Leu Val Cys Leu Ile
1055 1060 1065
Phe Trp Leu Ile Phe Ser Ile Met Gly Val Asn Leu Phe Ala Gly
1070 1075 1080
Arg Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg Phe Pro
1085 1090 1095
Ser Ser Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu Leu Phe
1100 1105 1110
Asn Glu Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe Asp Asn
1115 1120 1125
Val Gly Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr Phe Asn
1130 1135 1140
Gly Trp Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val Ala Val
1145 1150 1155
Asn Ile Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr Cys Tyr
1160 1165 1170
Phe Ile Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu Ser Met
1175 1180 1185
Leu Ile Thr Val Ile Ile Asp Asn Phe Asn Lys His Lys Ile Lys
1190 1195 1200
Leu Gly Gly Ser Asn Ile Phe Ile Thr Val Lys Gln Arg Lys Gln
1205 1210 1215
Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser Gln Arg Pro
1220 1225 1230
Val Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe Ile Phe Asp Val
1235 1240 1245
Val Thr Ser Gln Ala Phe Asn Val Ile Val Met Val Leu Ile Cys
1250 1255 1260
Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp Val Gln Ser Leu
1265 1270 1275
Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile Phe Val Met
1280 1285 1290
Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile Ala Phe Arg Cys
1295 1300 1305
Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe Met Val Val
1310 1315 1320
Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met Thr Val Gly Ser
1325 1330 1335
Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile Leu Leu Ser Arg
1340 1345 1350
Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro Lys Val Phe His
1355 1360 1365
Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro Ala Leu Leu Asn
1370 1375 1380
Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile Tyr Ala Val Phe
1385 1390 1395
Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala Gly Ile Asn
1400 1405 1410
Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser Met Leu Cys Leu
1415 1420 1425
Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly Met Leu Asp Ala
1430 1435 1440
Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp Lys Ile Asn
1445 1450 1455
Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn Pro Ser Val Gly
1460 1465 1470
Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp Leu Ile Ile
1475 1480 1485
Val Asn Met Tyr Ile Val Val Val Met Glu Phe Leu Asn Ile Ala
1490 1495 1500
Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp Phe Arg Lys
1505 1510 1515
Phe Phe Gln Val Trp Lys Arg Phe Asp Pro Asp Arg Thr Gln Tyr
1520 1525 1530
Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu Asp Pro
1535 1540 1545
Pro Leu Phe Met Ala Lys Pro Asn Lys Gly Gln Leu Ile Ala Leu
1550 1555 1560
Asp Leu Pro Met Ala Val Gly Asp Arg Ile His Cys Leu Asp Ile
1565 1570 1575
Leu Leu Ala Phe Thr Lys Arg Val Met Gly Gln Asp Val Arg Met
1580 1585 1590
Glu Lys Val Val Ser Glu Ile Glu Ser Gly Phe Leu Leu Ala Asn
1595 1600 1605
Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr Leu Lys Arg
1610 1615 1620
Lys Gln Glu Ala Val Ser Ala Thr Ile Ile Gln Arg Ala Tyr Lys
1625 1630 1635
Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser Asp Ile His
1640 1645 1650
Met Ile Asp Gly Asp Arg Asp Val His Ala Thr Lys Glu Gly Ala
1655 1660 1665
Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln Ser Gln Ile
1670 1675 1680
<210> 20
<211> 1730
<212> PRT
<213> 人工序列
<220>
<223> 合成的：hNaX_hNaV1.7_DIII_DIVS6-CTD_嵌合体
<400> 20
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys Thr Cys Tyr
915 920 925
Lys Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile Val Leu Met Ile
930 935 940
Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Arg
945 950 955 960
Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala Tyr Gly Tyr
980 985 990
Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val
995 1000 1005
Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu Gly Tyr Ser
1010 1015 1020
Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg
1025 1030 1035
Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val
1040 1045 1050
Asn Ala Leu Ile Gly Ala Ile Pro Ser Ile Met Asn Val Leu Leu
1055 1060 1065
Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile Met Gly Val Asn
1070 1075 1080
Leu Phe Ala Gly Lys Phe Tyr Glu Cys Ile Asn Thr Thr Asp Gly
1085 1090 1095
Ser Arg Phe Pro Ala Ser Gln Val Pro Asn Arg Ser Glu Cys Phe
1100 1105 1110
Ala Leu Met Asn Val Ser Gln Asn Val Arg Trp Lys Asn Leu Lys
1115 1120 1125
Val Asn Phe Asp Asn Val Gly Leu Gly Tyr Leu Ser Leu Leu Gln
1130 1135 1140
Val Ala Thr Phe Lys Gly Trp Thr Ile Ile Met Tyr Ala Ala Val
1145 1150 1155
Asp Ser Val Asn Val Asp Lys Gln Pro Lys Tyr Glu Tyr Ser Leu
1160 1165 1170
Tyr Met Tyr Ile Tyr Phe Val Val Phe Ile Ile Phe Gly Ser Phe
1175 1180 1185
Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe Asn
1190 1195 1200
Gln Gln Lys Lys Lys Leu Gly Gly Gln Asp Ile Phe Met Thr Glu
1205 1210 1215
Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys
1220 1225 1230
Lys Pro Gln Lys Pro Ile Pro Arg Pro Gly Asn Lys Leu Gln Gly
1235 1240 1245
Phe Ile Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val
1250 1255 1260
Met Val Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr
1265 1270 1275
Asp Val Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn
1280 1285 1290
Ser Ile Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu
1295 1300 1305
Ile Ala Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe
1310 1315 1320
Asp Phe Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro
1325 1330 1335
Met Thr Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu
1340 1345 1350
Ile Leu Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly
1355 1360 1365
Pro Lys Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu
1370 1375 1380
Pro Ala Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe
1385 1390 1395
Ile Tyr Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys
1400 1405 1410
Glu Ala Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn
1415 1420 1425
Ser Met Leu Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp
1430 1435 1440
Gly Met Leu Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp
1445 1450 1455
Pro Asp Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly
1460 1465 1470
Asn Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile
1475 1480 1485
Ser Trp Leu Ile Ile Val Asn Met Tyr Ile Ala Val Ile Leu Glu
1490 1495 1500
Asn Phe Ser Val Ala Thr Glu Glu Ser Thr Glu Pro Leu Ser Glu
1505 1510 1515
Asp Asp Phe Glu Met Phe Tyr Glu Val Trp Glu Lys Phe Asp Pro
1520 1525 1530
Asp Ala Thr Gln Phe Ile Glu Phe Ser Lys Leu Ser Asp Phe Ala
1535 1540 1545
Ala Ala Leu Asp Pro Pro Leu Leu Ile Ala Lys Pro Asn Lys Val
1550 1555 1560
Gln Leu Ile Ala Met Asp Leu Pro Met Val Ser Gly Asp Arg Ile
1565 1570 1575
His Cys Leu Asp Ile Leu Phe Ala Phe Thr Lys Arg Val Leu Gly
1580 1585 1590
Glu Ser Gly Glu Met Asp Ser Leu Arg Ser Gln Met Glu Glu Arg
1595 1600 1605
Phe Met Ser Ala Asn Pro Ser Lys Val Ser Tyr Glu Pro Ile Thr
1610 1615 1620
Thr Thr Leu Lys Arg Lys Gln Glu Asp Val Ser Ala Thr Val Ile
1625 1630 1635
Gln Arg Ala Tyr Arg Arg Tyr Arg Leu Arg Gln Asn Val Lys Asn
1640 1645 1650
Ile Ser Ser Ile Tyr Ile Lys Asp Gly Asp Arg Asp Asp Asp Leu
1655 1660 1665
Leu Asn Lys Lys Asp Met Ala Phe Asp Asn Val Asn Glu Asn Ser
1670 1675 1680
Ser Pro Glu Lys Thr Asp Ala Thr Ser Ser Thr Thr Ser Pro Pro
1685 1690 1695
Ser Tyr Asp Ser Val Thr Lys Pro Asp Lys Glu Lys Tyr Glu Gln
1700 1705 1710
Asp Arg Thr Glu Lys Glu Asp Lys Gly Lys Asp Ser Lys Glu Ser
1715 1720 1725
Lys Lys
1730
<210> 21
<211> 1876
<212> PRT
<213> 人工序列
<220>
<223> 合成的：NaX_NaV1.7_D1-D2_接头_嵌合体
<400> 21
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Gln Lys Glu Leu Glu Phe Gln Gln Met
385 390 395 400
Leu Asp Arg Leu Lys Lys Glu Gln Glu Glu Ala Glu Ala Ile Ala Ala
405 410 415
Ala Ala Ala Glu Tyr Thr Ser Ile Arg Arg Ser Arg Ile Met Gly Leu
420 425 430
Ser Glu Ser Ser Ser Glu Thr Ser Lys Leu Ser Ser Lys Ser Ala Lys
435 440 445
Glu Arg Arg Asn Arg Arg Lys Lys Lys Asn Gln Lys Lys Leu Ser Ser
450 455 460
Gly Glu Glu Lys Gly Asp Ala Glu Lys Leu Ser Lys Ser Glu Ser Glu
465 470 475 480
Asp Ser Ile Arg Arg Lys Ser Phe His Leu Gly Val Glu Gly His Arg
485 490 495
Arg Ala His Glu Lys Arg Leu Ser Thr Pro Asn Gln Ser Pro Leu Ser
500 505 510
Ile Arg Gly Ser Leu Phe Ser Ala Arg Arg Ser Ser Arg Thr Ser Leu
515 520 525
Phe Ser Phe Lys Gly Arg Gly Arg Asp Ile Gly Ser Glu Thr Glu Phe
530 535 540
Ala Asp Asp Glu His Ser Ile Phe Gly Asp Asn Glu Ser Arg Arg Gly
545 550 555 560
Ser Leu Phe Val Pro His Arg Pro Gln Glu Arg Arg Ser Ser Asn Ile
565 570 575
Ser Gln Ala Ser Arg Ser Pro Pro Met Leu Pro Val Asn Gly Lys Met
580 585 590
His Ser Ala Val Asp Cys Asn Gly Val Val Ser Leu Val Asp Gly Arg
595 600 605
Ser Ala Leu Met Leu Pro Asn Gly Gln Leu Leu Pro Glu Val Ile Ile
610 615 620
Asp Lys Ala Thr Ser Asp Asp Ser Gly Thr Thr Asn Gln Ile His Lys
625 630 635 640
Lys Arg Arg Cys Ser Ser Tyr Leu Leu Ser Glu Asp Met Leu Asn Asp
645 650 655
Pro Asn Leu Arg Gln Arg Ala Met Ser Arg Ala Ser Ile Leu Thr Asn
660 665 670
Thr Val Glu Glu Leu Glu Glu Ser Arg Gln Lys Cys Pro Pro Trp Trp
675 680 685
Tyr Arg Phe Ala His Lys Phe Leu Ile Trp Asn Phe Thr Asp Leu Phe
690 695 700
Leu Ile Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His
705 710 715 720
Tyr Pro Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu
725 730 735
Val Phe Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala
740 745 750
Met His Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser
755 760 765
Met Ile Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala
770 775 780
Gly Met Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu
785 790 795 800
Gly Lys Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn
805 810 815
Ser Trp Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile
820 825 830
Phe Phe Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu
835 840 845
Glu Phe Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His
850 855 860
Met His Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys
865 870 875 880
Gly Glu Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln
885 890 895
Ser Trp Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu
900 905 910
Leu Val Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys
915 920 925
Lys Asp Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu
930 935 940
Ala Val Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile
945 950 955 960
Leu Cys Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn
965 970 975
Glu Val Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu
980 985 990
Ser Asn Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr
995 1000 1005
Glu Lys Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser
1010 1015 1020
Pro Ser Val Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp
1025 1030 1035
Ile Glu Asn Leu Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp
1040 1045 1050
Gly Gly Ser Lys Glu Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys
1055 1060 1065
Ser Thr Val Asp Ile Ala Ile Ser Glu Glu Glu Glu Met Phe Tyr
1070 1075 1080
Gly Gly Glu Arg Ser Lys His Leu Lys Asn Gly Cys Arg Arg Gly
1085 1090 1095
Ser Ser Leu Gly Gln Ile Ser Gly Ala Ser Lys Lys Gly Lys Ile
1100 1105 1110
Trp Gln Asn Ile Arg Lys Thr Cys Cys Lys Ile Val Glu Asn Asn
1115 1120 1125
Trp Phe Lys Cys Phe Ile Gly Leu Val Thr Leu Leu Ser Thr Gly
1130 1135 1140
Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln Arg Lys Thr Ile
1145 1150 1155
Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr Tyr Ile Phe
1160 1165 1170
Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe Lys Ala
1175 1180 1185
Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val Ile
1190 1195 1200
Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg Glu Glu Leu Lys
1205 1210 1215
Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu Ser
1220 1225 1230
Gln Phe Glu Arg Met Lys Val Val Val Arg Ala Leu Ile Lys Thr
1235 1240 1245
Thr Leu Pro Thr Leu Asn Val Phe Leu Val Cys Leu Met Ile Trp
1250 1255 1260
Leu Ile Phe Ser Ile Met Gly Val Asp Leu Phe Ala Gly Arg Phe
1265 1270 1275
Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg Phe Pro Ser Ser
1280 1285 1290
Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu Leu Phe Asn Glu
1295 1300 1305
Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe Asp Asn Val Gly
1310 1315 1320
Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr Phe Asn Gly Trp
1325 1330 1335
Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val Ala Val Asn Ile
1340 1345 1350
Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr Cys Tyr Phe Ile
1355 1360 1365
Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu Ser Met Leu Ile
1370 1375 1380
Thr Val Ile Ile Asp Asn Phe Asn Lys His Lys Ile Lys Leu Gly
1385 1390 1395
Gly Ser Asn Ile Phe Ile Thr Val Lys Gln Arg Lys Gln Tyr Arg
1400 1405 1410
Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser Gln Arg Pro Val Pro
1415 1420 1425
Arg Pro Leu Asn Lys Leu Gln Gly Phe Ile Phe Asp Val Val Thr
1430 1435 1440
Ser Gln Ala Phe Asn Val Ile Val Met Val Leu Ile Cys Phe Gln
1445 1450 1455
Ala Ile Ala Met Met Ile Asp Thr Asp Val Gln Ser Leu Gln Met
1460 1465 1470
Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile Phe Val Met Leu Tyr
1475 1480 1485
Thr Met Glu Cys Ile Leu Lys Leu Ile Ala Phe Arg Cys Phe Tyr
1490 1495 1500
Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe Met Val Val Ile Phe
1505 1510 1515
Ser Ile Thr Gly Leu Cys Leu Pro Met Thr Val Gly Ser Tyr Leu
1520 1525 1530
Val Pro Pro Ser Leu Val Gln Leu Ile Leu Leu Ser Arg Ile Ile
1535 1540 1545
His Met Leu Arg Leu Gly Lys Gly Pro Lys Val Phe His Asn Leu
1550 1555 1560
Met Leu Pro Leu Met Leu Ser Leu Pro Ala Leu Leu Asn Ile Ile
1565 1570 1575
Leu Leu Ile Phe Leu Val Met Phe Ile Tyr Ala Val Phe Gly Met
1580 1585 1590
Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala Gly Ile Asn Asp Val
1595 1600 1605
Ser Asn Phe Glu Thr Phe Gly Asn Ser Met Leu Cys Leu Phe Gln
1610 1615 1620
Val Ala Ile Phe Ala Gly Trp Asp Gly Met Leu Asp Ala Ile Phe
1625 1630 1635
Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp Lys Ile Asn Pro Gly
1640 1645 1650
Thr Gln Val Arg Gly Asp Cys Gly Asn Pro Ser Val Gly Ile Phe
1655 1660 1665
Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp Leu Ile Ile Val Asn
1670 1675 1680
Met Tyr Ile Val Val Val Met Glu Phe Leu Asn Ile Ala Ser Lys
1685 1690 1695
Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp Phe Arg Lys Phe Phe
1700 1705 1710
Gln Val Trp Lys Arg Phe Asp Pro Asp Arg Thr Gln Tyr Ile Asp
1715 1720 1725
Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu Asp Pro Pro Leu
1730 1735 1740
Phe Met Ala Lys Pro Asn Lys Gly Gln Leu Ile Ala Leu Asp Leu
1745 1750 1755
Pro Met Ala Val Gly Asp Arg Ile His Cys Leu Asp Ile Leu Leu
1760 1765 1770
Ala Phe Thr Lys Arg Val Met Gly Gln Asp Val Arg Met Glu Lys
1775 1780 1785
Val Val Ser Glu Ile Glu Ser Gly Phe Leu Leu Ala Asn Pro Phe
1790 1795 1800
Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr Leu Lys Arg Lys Gln
1805 1810 1815
Glu Ala Val Ser Ala Thr Ile Ile Gln Arg Ala Tyr Lys Asn Tyr
1820 1825 1830
Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser Asp Ile His Met Ile
1835 1840 1845
Asp Gly Asp Arg Asp Val His Ala Thr Lys Glu Gly Ala Tyr Phe
1850 1855 1860
Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln Ser Gln Ile
1865 1870 1875
<210> 22
<211> 1701
<212> PRT
<213> 人工序列
<220>
<223> 合成的：NaX_NaV1.7_D2-D3_接头_嵌合体
<400> 22
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Ser Phe Ser Ser Asp Asn Leu Thr Ala
725 730 735
Ile Glu Glu Asp Pro Asp Ala Asn Asn Leu Gln Ile Ala Val Thr Arg
740 745 750
Ile Lys Lys Gly Ile Asn Tyr Val Lys Gln Thr Leu Arg Glu Phe Ile
755 760 765
Leu Lys Ala Phe Ser Lys Lys Pro Lys Ile Ser Arg Glu Ile Arg Gln
770 775 780
Ala Glu Asp Leu Asn Thr Lys Lys Glu Asn Tyr Ile Ser Asn His Thr
785 790 795 800
Leu Ala Glu Met Ser Lys Gly His Asn Phe Leu Lys Glu Lys Asp Lys
805 810 815
Ile Ser Gly Phe Gly Ser Ser Val Asp Lys His Leu Met Glu Asp Ser
820 825 830
Asp Gly Gln Ser Phe Ile His Asn Pro Ser Leu Thr Val Thr Val Pro
835 840 845
Ile Ala Pro Gly Glu Ser Asp Leu Glu Asn Met Asn Ala Glu Glu Leu
850 855 860
Ser Ser Asp Ser Asp Ser Glu Tyr Ser Lys Val Arg Leu Asn Arg Ser
865 870 875 880
Ser Ser Ser Glu Cys Ser Thr Val Asp Asn Pro Leu Pro Gly Glu Gly
885 890 895
Glu Glu Ala Glu Ala Glu Pro Met Asn Ser Asp Glu Pro Glu Ala Cys
900 905 910
Phe Thr Asp Gly Cys Val Trp Arg Phe Ser Cys Cys Gln Val Asn Ile
915 920 925
Glu Ser Gly Lys Gly Lys Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys
930 935 940
Thr Cys Cys Lys Ile Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly
945 950 955 960
Leu Val Thr Leu Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr
965 970 975
Met Asp Gln Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met
980 985 990
Ile Phe Thr Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala
995 1000 1005
Tyr Gly Phe Lys Ala Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp
1010 1015 1020
Phe Val Val Val Ile Val Phe Cys Leu Ser Leu Ile Gly Lys Thr
1025 1030 1035
Arg Glu Glu Leu Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro
1040 1045 1050
Leu Arg Val Leu Ser Gln Phe Glu Arg Met Lys Val Val Val Arg
1055 1060 1065
Ala Leu Ile Lys Thr Thr Leu Pro Thr Leu Asn Val Phe Leu Val
1070 1075 1080
Cys Leu Met Ile Trp Leu Ile Phe Ser Ile Met Gly Val Asp Leu
1085 1090 1095
Phe Ala Gly Arg Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu
1100 1105 1110
Arg Phe Pro Ser Ser Glu Val Met Asn Lys Ser Arg Cys Glu Ser
1115 1120 1125
Leu Leu Phe Asn Glu Ser Met Leu Trp Glu Asn Ala Lys Met Asn
1130 1135 1140
Phe Asp Asn Val Gly Asn Gly Phe Leu Ser Leu Leu Gln Val Ala
1145 1150 1155
Thr Phe Asn Gly Trp Ile Thr Ile Met Asn Ser Ala Ile Asp Ser
1160 1165 1170
Val Ala Val Asn Ile Gln Pro His Phe Glu Val Asn Ile Tyr Met
1175 1180 1185
Tyr Cys Tyr Phe Ile Asn Phe Ile Ile Phe Gly Val Phe Leu Pro
1190 1195 1200
Leu Ser Met Leu Ile Thr Val Ile Ile Asp Asn Phe Asn Lys His
1205 1210 1215
Lys Ile Lys Leu Gly Gly Ser Asn Ile Phe Ile Thr Val Lys Gln
1220 1225 1230
Arg Lys Gln Tyr Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser
1235 1240 1245
Gln Arg Pro Val Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe Ile
1250 1255 1260
Phe Asp Val Val Thr Ser Gln Ala Phe Asn Val Ile Val Met Val
1265 1270 1275
Leu Ile Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp Val
1280 1285 1290
Gln Ser Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile
1295 1300 1305
Phe Val Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile Ala
1310 1315 1320
Phe Arg Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe
1325 1330 1335
Met Val Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met Thr
1340 1345 1350
Val Gly Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile Leu
1355 1360 1365
Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro Lys
1370 1375 1380
Val Phe His Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro Ala
1385 1390 1395
Leu Leu Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile Tyr
1400 1405 1410
Ala Val Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala
1415 1420 1425
Gly Ile Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser Met
1430 1435 1440
Leu Cys Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly Met
1445 1450 1455
Leu Asp Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp
1460 1465 1470
Lys Ile Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn Pro
1475 1480 1485
Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp
1490 1495 1500
Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu Phe Leu
1505 1510 1515
Asn Ile Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp
1520 1525 1530
Phe Arg Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro Asp Arg
1535 1540 1545
Thr Gln Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala
1550 1555 1560
Leu Asp Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly Gln Leu
1565 1570 1575
Ile Ala Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile His Cys
1580 1585 1590
Leu Asp Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly Gln Asp
1595 1600 1605
Val Arg Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly Phe Leu
1610 1615 1620
Leu Ala Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr
1625 1630 1635
Leu Lys Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile Gln Arg
1640 1645 1650
Ala Tyr Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser
1655 1660 1665
Asp Ile His Met Ile Asp Gly Asp Arg Asp Val His Ala Thr Lys
1670 1675 1680
Glu Gly Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln
1685 1690 1695
Ser Gln Ile
1700
<210> 23
<211> 1669
<212> PRT
<213> 人工序列
<220>
<223> 合成的：NaX_NaV1.7_D3-D4_接头_嵌合体
<400> 23
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys Thr Cys Cys
915 920 925
Lys Ile Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly Leu Val Thr
930 935 940
Leu Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln
945 950 955 960
Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe
980 985 990
Lys Ala Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val
995 1000 1005
Ile Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg Glu Glu Leu
1010 1015 1020
Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu
1025 1030 1035
Ser Gln Phe Glu Arg Met Lys Val Val Val Arg Ala Leu Ile Lys
1040 1045 1050
Thr Thr Leu Pro Thr Leu Asn Val Phe Leu Val Cys Leu Met Ile
1055 1060 1065
Trp Leu Ile Phe Ser Ile Met Gly Val Asp Leu Phe Ala Gly Arg
1070 1075 1080
Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg Phe Pro Ser
1085 1090 1095
Ser Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu Leu Phe Asn
1100 1105 1110
Glu Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe Asp Asn Val
1115 1120 1125
Gly Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr Phe Asn Gly
1130 1135 1140
Trp Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val Ala Val Asn
1145 1150 1155
Ile Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr Cys Tyr Phe
1160 1165 1170
Ile Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu Ser Met Leu
1175 1180 1185
Ile Leu Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys
1190 1195 1200
Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys
1205 1210 1215
Pro Ile Pro Arg Pro Gly Asn Lys Leu Gln Gly Phe Ile Phe Asp
1220 1225 1230
Val Val Thr Ser Gln Ala Phe Asn Val Ile Val Met Val Leu Ile
1235 1240 1245
Cys Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp Val Gln Ser
1250 1255 1260
Leu Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile Phe Val
1265 1270 1275
Met Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile Ala Phe Arg
1280 1285 1290
Cys Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe Met Val
1295 1300 1305
Val Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met Thr Val Gly
1310 1315 1320
Ser Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile Leu Leu Ser
1325 1330 1335
Arg Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro Lys Val Phe
1340 1345 1350
His Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro Ala Leu Leu
1355 1360 1365
Asn Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile Tyr Ala Val
1370 1375 1380
Phe Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala Gly Ile
1385 1390 1395
Asn Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser Met Leu Cys
1400 1405 1410
Leu Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly Met Leu Asp
1415 1420 1425
Ala Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp Lys Ile
1430 1435 1440
Asn Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn Pro Ser Val
1445 1450 1455
Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp Leu Ile
1460 1465 1470
Ile Val Asn Met Tyr Ile Val Val Val Met Glu Phe Leu Asn Ile
1475 1480 1485
Ala Ser Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp Phe Arg
1490 1495 1500
Lys Phe Phe Gln Val Trp Lys Arg Phe Asp Pro Asp Arg Thr Gln
1505 1510 1515
Tyr Ile Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu Asp
1520 1525 1530
Pro Pro Leu Phe Met Ala Lys Pro Asn Lys Gly Gln Leu Ile Ala
1535 1540 1545
Leu Asp Leu Pro Met Ala Val Gly Asp Arg Ile His Cys Leu Asp
1550 1555 1560
Ile Leu Leu Ala Phe Thr Lys Arg Val Met Gly Gln Asp Val Arg
1565 1570 1575
Met Glu Lys Val Val Ser Glu Ile Glu Ser Gly Phe Leu Leu Ala
1580 1585 1590
Asn Pro Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr Leu Lys
1595 1600 1605
Arg Lys Gln Glu Ala Val Ser Ala Thr Ile Ile Gln Arg Ala Tyr
1610 1615 1620
Lys Asn Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser Asp Ile
1625 1630 1635
His Met Ile Asp Gly Asp Arg Asp Val His Ala Thr Lys Glu Gly
1640 1645 1650
Ala Tyr Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln Ser Gln
1655 1660 1665
Ile
<210> 24
<211> 1911
<212> PRT
<213> 人工序列
<220>
<223> 合成的：NaX_NaV1.7_最终环_嵌合体
<400> 24
Met Ala Met Leu Pro Pro Pro Gly Pro Gln Ser Phe Val His Phe Thr
1 5 10 15
Lys Gln Ser Leu Ala Leu Ile Glu Gln Arg Ile Ala Glu Arg Lys Ser
20 25 30
Lys Glu Pro Lys Glu Glu Lys Lys Asp Asp Asp Glu Glu Ala Pro Lys
35 40 45
Pro Ser Ser Asp Leu Glu Ala Gly Lys Gln Leu Pro Phe Ile Tyr Gly
50 55 60
Asp Ile Pro Pro Gly Met Val Ser Glu Pro Leu Glu Asp Leu Asp Pro
65 70 75 80
Tyr Tyr Ala Asp Lys Lys Thr Phe Ile Val Leu Asn Lys Gly Lys Thr
85 90 95
Ile Phe Arg Phe Asn Ala Thr Pro Ala Leu Tyr Met Leu Ser Pro Phe
100 105 110
Ser Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val Leu Ile Asp Cys
115 120 125
Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg Pro Val Leu Glu
130 135 140
Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu Val Lys Leu Phe
145 150 155 160
Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu Gly Asp Pro Trp
165 170 175
Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val Ile Ile Arg Tyr
180 185 190
Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala Arg Thr Leu Arg
195 200 205
Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys Ser Leu Val Gly
210 215 220
Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val Ile Ile Leu Thr
225 230 235 240
Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met Gly Leu Phe Met
245 250 255
Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln Glu Asn Glu Asn
260 265 270
Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr Ile Arg Glu Thr
275 280 285
Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala Leu Leu Cys Gly
290 295 300
Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr Val Cys Val Lys
305 310 315 320
Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe Asp Ser Phe Gly
325 330 335
Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln Asp Tyr Pro Glu
340 345 350
Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys Val Tyr Met Ile
355 360 365
Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr Met Ala Ser Leu
370 375 380
Phe Leu Gly Gln Lys Glu Leu Glu Phe Gln Gln Met Leu Asp Arg Leu
385 390 395 400
Lys Lys Glu Gln Glu Glu Ala Glu Ala Ile Ala Ala Ala Ala Ala Glu
405 410 415
Tyr Thr Ser Ile Arg Arg Ser Arg Ile Met Gly Leu Ser Glu Ser Ser
420 425 430
Ser Glu Thr Ser Lys Leu Ser Ser Lys Ser Ala Lys Glu Arg Arg Asn
435 440 445
Arg Arg Lys Lys Lys Asn Gln Lys Lys Leu Ser Ser Gly Glu Glu Lys
450 455 460
Gly Asp Ala Glu Lys Leu Ser Lys Ser Glu Ser Glu Asp Ser Ile Arg
465 470 475 480
Arg Lys Ser Phe His Leu Gly Val Glu Gly His Arg Arg Ala His Glu
485 490 495
Lys Arg Leu Ser Thr Pro Asn Gln Ser Pro Leu Ser Ile Arg Gly Ser
500 505 510
Leu Phe Ser Ala Arg Arg Ser Ser Arg Thr Ser Leu Phe Ser Phe Lys
515 520 525
Gly Arg Gly Arg Asp Ile Gly Ser Glu Thr Glu Phe Ala Asp Asp Glu
530 535 540
His Ser Ile Phe Gly Asp Asn Glu Ser Arg Arg Gly Ser Leu Phe Val
545 550 555 560
Pro His Arg Pro Gln Glu Arg Arg Ser Ser Asn Ile Ser Gln Ala Ser
565 570 575
Arg Ser Pro Pro Met Leu Pro Val Asn Gly Lys Met His Ser Ala Val
580 585 590
Asp Cys Asn Gly Val Val Ser Leu Val Asp Gly Arg Ser Ala Leu Met
595 600 605
Leu Pro Asn Gly Gln Leu Leu Pro Glu Val Ile Ile Asp Lys Ala Thr
610 615 620
Ser Asp Asp Ser Gly Thr Thr Asn Gln Ile His Lys Lys Arg Arg Cys
625 630 635 640
Ser Ser Tyr Leu Leu Ser Glu Asp Met Leu Asn Asp Pro Asn Leu Arg
645 650 655
Gln Arg Ala Met Ser Arg Ala Ser Ile Leu Thr Asn Thr Val Glu Glu
660 665 670
Leu Glu Glu Ser Arg Gln Lys Cys Pro Pro Trp Trp Tyr Arg Phe Ala
675 680 685
His Lys Phe Leu Ile Trp Asn Phe Thr Asp Leu Phe Leu Ile Ile Cys
690 695 700
Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro Met Ser
705 710 715 720
Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe Ile Gly
725 730 735
Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His Pro Tyr
740 745 750
Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile Val Phe
755 760 765
His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met Ala Leu
770 775 780
Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys Tyr Trp
785 790 795 800
Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp Val Ala
805 810 815
Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe Ser Ala
820 825 830
Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe Val Cys
835 840 845
His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His Asp Phe
850 855 860
Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu Trp Val
865 870 875 880
Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp Cys Ile
885 890 895
Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val Leu Tyr
900 905 910
Leu Phe Leu Ala Leu Ser Phe Ser Ser Asp Asn Leu Thr Ala Ile Glu
915 920 925
Glu Asp Pro Asp Ala Asn Asn Leu Gln Ile Ala Val Thr Arg Ile Lys
930 935 940
Lys Gly Ile Asn Tyr Val Lys Gln Thr Leu Arg Glu Phe Ile Leu Lys
945 950 955 960
Ala Phe Ser Lys Lys Pro Lys Ile Ser Arg Glu Ile Arg Gln Ala Glu
965 970 975
Asp Leu Asn Thr Lys Lys Glu Asn Tyr Ile Ser Asn His Thr Leu Ala
980 985 990
Glu Met Ser Lys Gly His Asn Phe Leu Lys Glu Lys Asp Lys Ile Ser
995 1000 1005
Gly Phe Gly Ser Ser Val Asp Lys His Leu Met Glu Asp Ser Asp
1010 1015 1020
Gly Gln Ser Phe Ile His Asn Pro Ser Leu Thr Val Thr Val Pro
1025 1030 1035
Ile Ala Pro Gly Glu Ser Asp Leu Glu Asn Met Asn Ala Glu Glu
1040 1045 1050
Leu Ser Ser Asp Ser Asp Ser Glu Tyr Ser Lys Val Arg Leu Asn
1055 1060 1065
Arg Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Asn Pro Leu Pro
1070 1075 1080
Gly Glu Gly Glu Glu Ala Glu Ala Glu Pro Met Asn Ser Asp Glu
1085 1090 1095
Pro Glu Ala Cys Phe Thr Asp Gly Cys Val Trp Arg Phe Ser Cys
1100 1105 1110
Cys Gln Val Asn Ile Glu Ser Gly Lys Gly Lys Lys Gly Lys Ile
1115 1120 1125
Trp Gln Asn Ile Arg Lys Thr Cys Cys Lys Ile Val Glu Asn Asn
1130 1135 1140
Trp Phe Lys Cys Phe Ile Gly Leu Val Thr Leu Leu Ser Thr Gly
1145 1150 1155
Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln Arg Lys Thr Ile
1160 1165 1170
Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr Tyr Ile Phe
1175 1180 1185
Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe Lys Ala
1190 1195 1200
Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val Ile
1205 1210 1215
Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg Glu Glu Leu Lys
1220 1225 1230
Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu Ser
1235 1240 1245
Gln Phe Glu Arg Met Lys Val Val Val Arg Ala Leu Ile Lys Thr
1250 1255 1260
Thr Leu Pro Thr Leu Asn Val Phe Leu Val Cys Leu Met Ile Trp
1265 1270 1275
Leu Ile Phe Ser Ile Met Gly Val Asp Leu Phe Ala Gly Arg Phe
1280 1285 1290
Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg Phe Pro Ser Ser
1295 1300 1305
Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu Leu Phe Asn Glu
1310 1315 1320
Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe Asp Asn Val Gly
1325 1330 1335
Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr Phe Asn Gly Trp
1340 1345 1350
Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val Ala Val Asn Ile
1355 1360 1365
Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr Cys Tyr Phe Ile
1370 1375 1380
Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu Ser Met Leu Ile
1385 1390 1395
Leu Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr
1400 1405 1410
Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro
1415 1420 1425
Ile Pro Arg Pro Gly Asn Lys Leu Gln Gly Phe Ile Phe Asp Val
1430 1435 1440
Val Thr Ser Gln Ala Phe Asn Val Ile Val Met Val Leu Ile Cys
1445 1450 1455
Phe Gln Ala Ile Ala Met Met Ile Asp Thr Asp Val Gln Ser Leu
1460 1465 1470
Gln Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile Phe Val Met
1475 1480 1485
Leu Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile Ala Phe Arg Cys
1490 1495 1500
Phe Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe Met Val Val
1505 1510 1515
Ile Phe Ser Ile Thr Gly Leu Cys Leu Pro Met Thr Val Gly Ser
1520 1525 1530
Tyr Leu Val Pro Pro Ser Leu Val Gln Leu Ile Leu Leu Ser Arg
1535 1540 1545
Ile Ile His Met Leu Arg Leu Gly Lys Gly Pro Lys Val Phe His
1550 1555 1560
Asn Leu Met Leu Pro Leu Met Leu Ser Leu Pro Ala Leu Leu Asn
1565 1570 1575
Ile Ile Leu Leu Ile Phe Leu Val Met Phe Ile Tyr Ala Val Phe
1580 1585 1590
Gly Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala Gly Ile Asn
1595 1600 1605
Asp Val Ser Asn Phe Glu Thr Phe Gly Asn Ser Met Leu Cys Leu
1610 1615 1620
Phe Gln Val Ala Ile Phe Ala Gly Trp Asp Gly Met Leu Asp Ala
1625 1630 1635
Ile Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp Lys Ile Asn
1640 1645 1650
Pro Gly Thr Gln Val Arg Gly Asp Cys Gly Asn Pro Ser Val Gly
1655 1660 1665
Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp Leu Ile Ile
1670 1675 1680
Val Asn Met Tyr Ile Val Val Val Glu Ser Thr Glu Pro Leu Ser
1685 1690 1695
Glu Asp Asp Phe Glu Met Phe Tyr Glu Val Trp Glu Lys Phe Asp
1700 1705 1710
Pro Asp Ala Thr Gln Phe Ile Glu Phe Ser Lys Leu Ser Asp Phe
1715 1720 1725
Ala Ala Ala Leu Asp Pro Pro Leu Leu Ile Ala Lys Pro Asn Lys
1730 1735 1740
Val Gln Leu Ile Ala Met Asp Leu Pro Met Val Ser Gly Asp Arg
1745 1750 1755
Ile His Cys Leu Asp Ile Leu Phe Ala Phe Thr Lys Arg Val Leu
1760 1765 1770
Gly Glu Ser Gly Glu Met Asp Ser Leu Arg Ser Gln Met Glu Glu
1775 1780 1785
Arg Phe Met Ser Ala Asn Pro Ser Lys Val Ser Tyr Glu Pro Ile
1790 1795 1800
Thr Thr Thr Leu Lys Arg Lys Gln Glu Asp Val Ser Ala Thr Val
1805 1810 1815
Ile Gln Arg Ala Tyr Arg Arg Tyr Arg Leu Arg Gln Asn Val Lys
1820 1825 1830
Asn Ile Ser Ser Ile Tyr Ile Lys Asp Gly Asp Arg Asp Asp Asp
1835 1840 1845
Leu Leu Asn Lys Lys Asp Met Ala Phe Asp Asn Val Asn Glu Asn
1850 1855 1860
Ser Ser Pro Glu Lys Thr Asp Ala Thr Ser Ser Thr Thr Ser Pro
1865 1870 1875
Pro Ser Tyr Asp Ser Val Thr Lys Pro Asp Lys Glu Lys Tyr Glu
1880 1885 1890
Gln Asp Arg Thr Glu Lys Glu Asp Lys Gly Lys Asp Ser Lys Glu
1895 1900 1905
Ser Lys Lys
1910
<210> 25
<211> 1682
<212> PRT
<213> 人工序列
<220>
<223> 合成的：NaX_QTT
<400> 25
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Gln Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys Thr Cys Cys
915 920 925
Lys Ile Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly Leu Val Thr
930 935 940
Leu Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln
945 950 955 960
Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe
980 985 990
Lys Ala Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val
995 1000 1005
Ile Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg Glu Glu Leu
1010 1015 1020
Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu
1025 1030 1035
Ser Gln Phe Glu Arg Met Lys Val Val Val Arg Ala Leu Ile Lys
1040 1045 1050
Thr Thr Leu Pro Thr Leu Asn Val Phe Leu Val Cys Leu Met Ile
1055 1060 1065
Trp Leu Ile Phe Ser Ile Met Gly Val Asp Leu Phe Ala Gly Arg
1070 1075 1080
Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg Phe Pro Ser
1085 1090 1095
Ser Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu Leu Phe Asn
1100 1105 1110
Glu Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe Asp Asn Val
1115 1120 1125
Gly Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr Phe Asn Gly
1130 1135 1140
Trp Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val Ala Val Asn
1145 1150 1155
Ile Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr Cys Tyr Phe
1160 1165 1170
Ile Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu Ser Met Leu
1175 1180 1185
Thr Thr Val Ile Ile Asp Asn Phe Asn Lys His Lys Ile Lys Leu
1190 1195 1200
Gly Gly Ser Asn Ile Phe Ile Thr Val Lys Gln Arg Lys Gln Tyr
1205 1210 1215
Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser Gln Arg Pro Val
1220 1225 1230
Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe Ile Phe Asp Val Val
1235 1240 1245
Thr Ser Gln Ala Phe Asn Val Ile Val Met Val Leu Ile Cys Phe
1250 1255 1260
Gln Ala Ile Ala Met Met Ile Asp Thr Asp Val Gln Ser Leu Gln
1265 1270 1275
Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile Phe Val Met Leu
1280 1285 1290
Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile Ala Phe Arg Cys Phe
1295 1300 1305
Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe Met Val Val Ile
1310 1315 1320
Phe Ser Ile Thr Gly Leu Cys Leu Pro Met Thr Val Gly Ser Tyr
1325 1330 1335
Leu Val Pro Pro Ser Leu Val Gln Leu Ile Leu Leu Ser Arg Ile
1340 1345 1350
Ile His Met Leu Arg Leu Gly Lys Gly Pro Lys Val Phe His Asn
1355 1360 1365
Leu Met Leu Pro Leu Met Leu Ser Leu Pro Ala Leu Leu Asn Ile
1370 1375 1380
Ile Leu Leu Ile Phe Leu Val Met Phe Ile Tyr Ala Val Phe Gly
1385 1390 1395
Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala Gly Ile Asn Asp
1400 1405 1410
Val Ser Asn Phe Glu Thr Phe Gly Asn Ser Met Leu Cys Leu Phe
1415 1420 1425
Gln Val Ala Ile Phe Ala Gly Trp Asp Gly Met Leu Asp Ala Ile
1430 1435 1440
Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp Lys Ile Asn Pro
1445 1450 1455
Gly Thr Gln Val Arg Gly Asp Cys Gly Asn Pro Ser Val Gly Ile
1460 1465 1470
Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp Leu Ile Ile Val
1475 1480 1485
Asn Met Tyr Thr Val Val Val Met Glu Phe Leu Asn Ile Ala Ser
1490 1495 1500
Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp Phe Arg Lys Phe
1505 1510 1515
Phe Gln Val Trp Lys Arg Phe Asp Pro Asp Arg Thr Gln Tyr Ile
1520 1525 1530
Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu Asp Pro Pro
1535 1540 1545
Leu Phe Met Ala Lys Pro Asn Lys Gly Gln Leu Ile Ala Leu Asp
1550 1555 1560
Leu Pro Met Ala Val Gly Asp Arg Ile His Cys Leu Asp Ile Leu
1565 1570 1575
Leu Ala Phe Thr Lys Arg Val Met Gly Gln Asp Val Arg Met Glu
1580 1585 1590
Lys Val Val Ser Glu Ile Glu Ser Gly Phe Leu Leu Ala Asn Pro
1595 1600 1605
Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr Leu Lys Arg Lys
1610 1615 1620
Gln Glu Ala Val Ser Ala Thr Ile Ile Gln Arg Ala Tyr Lys Asn
1625 1630 1635
Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser Asp Ile His Met
1640 1645 1650
Ile Asp Gly Asp Arg Asp Val His Ala Thr Lys Glu Gly Ala Tyr
1655 1660 1665
Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln Ser Gln Ile
1670 1675 1680
<210> 26
<211> 1682
<212> PRT
<213> 人工序列
<220>
<223> 合成的：NaX_EEE
<400> 26
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Glu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys Thr Cys Cys
915 920 925
Lys Ile Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly Leu Val Thr
930 935 940
Leu Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln
945 950 955 960
Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe
980 985 990
Lys Ala Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val
995 1000 1005
Ile Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg Glu Glu Leu
1010 1015 1020
Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu
1025 1030 1035
Ser Gln Phe Glu Arg Met Lys Val Val Val Arg Ala Leu Ile Lys
1040 1045 1050
Thr Thr Leu Pro Thr Leu Asn Val Phe Leu Val Cys Leu Met Ile
1055 1060 1065
Trp Leu Ile Phe Ser Ile Met Gly Val Asp Leu Phe Ala Gly Arg
1070 1075 1080
Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg Phe Pro Ser
1085 1090 1095
Ser Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu Leu Phe Asn
1100 1105 1110
Glu Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe Asp Asn Val
1115 1120 1125
Gly Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr Phe Asn Gly
1130 1135 1140
Trp Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val Ala Val Asn
1145 1150 1155
Ile Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr Cys Tyr Phe
1160 1165 1170
Ile Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu Ser Met Leu
1175 1180 1185
Glu Thr Val Ile Ile Asp Asn Phe Asn Lys His Lys Ile Lys Leu
1190 1195 1200
Gly Gly Ser Asn Ile Phe Ile Thr Val Lys Gln Arg Lys Gln Tyr
1205 1210 1215
Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser Gln Arg Pro Val
1220 1225 1230
Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe Ile Phe Asp Val Val
1235 1240 1245
Thr Ser Gln Ala Phe Asn Val Ile Val Met Val Leu Ile Cys Phe
1250 1255 1260
Gln Ala Ile Ala Met Met Ile Asp Thr Asp Val Gln Ser Leu Gln
1265 1270 1275
Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile Phe Val Met Leu
1280 1285 1290
Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile Ala Phe Arg Cys Phe
1295 1300 1305
Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe Met Val Val Ile
1310 1315 1320
Phe Ser Ile Thr Gly Leu Cys Leu Pro Met Thr Val Gly Ser Tyr
1325 1330 1335
Leu Val Pro Pro Ser Leu Val Gln Leu Ile Leu Leu Ser Arg Ile
1340 1345 1350
Ile His Met Leu Arg Leu Gly Lys Gly Pro Lys Val Phe His Asn
1355 1360 1365
Leu Met Leu Pro Leu Met Leu Ser Leu Pro Ala Leu Leu Asn Ile
1370 1375 1380
Ile Leu Leu Ile Phe Leu Val Met Phe Ile Tyr Ala Val Phe Gly
1385 1390 1395
Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala Gly Ile Asn Asp
1400 1405 1410
Val Ser Asn Phe Glu Thr Phe Gly Asn Ser Met Leu Cys Leu Phe
1415 1420 1425
Gln Val Ala Ile Phe Ala Gly Trp Asp Gly Met Leu Asp Ala Ile
1430 1435 1440
Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp Lys Ile Asn Pro
1445 1450 1455
Gly Thr Gln Val Arg Gly Asp Cys Gly Asn Pro Ser Val Gly Ile
1460 1465 1470
Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp Leu Ile Ile Val
1475 1480 1485
Asn Met Tyr Glu Val Val Val Met Glu Phe Leu Asn Ile Ala Ser
1490 1495 1500
Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp Phe Arg Lys Phe
1505 1510 1515
Phe Gln Val Trp Lys Arg Phe Asp Pro Asp Arg Thr Gln Tyr Ile
1520 1525 1530
Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu Asp Pro Pro
1535 1540 1545
Leu Phe Met Ala Lys Pro Asn Lys Gly Gln Leu Ile Ala Leu Asp
1550 1555 1560
Leu Pro Met Ala Val Gly Asp Arg Ile His Cys Leu Asp Ile Leu
1565 1570 1575
Leu Ala Phe Thr Lys Arg Val Met Gly Gln Asp Val Arg Met Glu
1580 1585 1590
Lys Val Val Ser Glu Ile Glu Ser Gly Phe Leu Leu Ala Asn Pro
1595 1600 1605
Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr Leu Lys Arg Lys
1610 1615 1620
Gln Glu Ala Val Ser Ala Thr Ile Ile Gln Arg Ala Tyr Lys Asn
1625 1630 1635
Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser Asp Ile His Met
1640 1645 1650
Ile Asp Gly Asp Arg Asp Val His Ala Thr Lys Glu Gly Ala Tyr
1655 1660 1665
Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln Ser Gln Ile
1670 1675 1680
<210> 27
<211> 1682
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(1682)
<223> 人 Nax（图 9）
<400> 27
Met Leu Ala Ser Pro Glu Pro Lys Gly Leu Val Pro Phe Thr Lys Glu
1 5 10 15
Ser Phe Glu Leu Ile Lys Gln His Ile Ala Lys Thr His Asn Glu Asp
20 25 30
His Glu Glu Glu Asp Leu Lys Pro Thr Pro Asp Leu Glu Val Gly Lys
35 40 45
Lys Leu Pro Phe Ile Tyr Gly Asn Leu Ser Gln Gly Met Val Ser Glu
50 55 60
Pro Leu Glu Asp Val Asp Pro Tyr Tyr Tyr Lys Lys Lys Asn Thr Phe
65 70 75 80
Ile Val Leu Asn Lys Asn Arg Thr Ile Phe Arg Phe Asn Ala Ala Ser
85 90 95
Ile Leu Cys Thr Leu Ser Pro Phe Asn Cys Ile Arg Arg Thr Thr Ile
100 105 110
Lys Val Leu Val His Pro Phe Phe Gln Leu Phe Ile Leu Ile Ser Val
115 120 125
Leu Ile Asp Cys Val Phe Met Ser Leu Thr Asn Leu Pro Lys Trp Arg
130 135 140
Pro Val Leu Glu Asn Thr Leu Leu Gly Ile Tyr Thr Phe Glu Ile Leu
145 150 155 160
Val Lys Leu Phe Ala Arg Gly Val Trp Ala Gly Ser Phe Ser Phe Leu
165 170 175
Gly Asp Pro Trp Asn Trp Leu Asp Phe Ser Val Thr Val Phe Glu Val
180 185 190
Ile Ile Arg Tyr Ser Pro Leu Asp Phe Ile Pro Thr Leu Gln Thr Ala
195 200 205
Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro Leu Asn Gln Gly Leu Lys
210 215 220
Ser Leu Val Gly Val Leu Ile His Cys Leu Lys Gln Leu Ile Gly Val
225 230 235 240
Ile Ile Leu Thr Leu Phe Phe Leu Ser Ile Phe Ser Leu Ile Gly Met
245 250 255
Gly Leu Phe Met Gly Asn Leu Lys His Lys Cys Phe Arg Trp Pro Gln
260 265 270
Glu Asn Glu Asn Glu Thr Leu His Asn Arg Thr Gly Asn Pro Tyr Tyr
275 280 285
Ile Arg Glu Thr Glu Asn Phe Tyr Tyr Leu Glu Gly Glu Arg Tyr Ala
290 295 300
Leu Leu Cys Gly Asn Arg Thr Asp Ala Gly Gln Cys Pro Glu Gly Tyr
305 310 315 320
Val Cys Val Lys Ala Gly Ile Asn Pro Asp Gln Gly Phe Thr Asn Phe
325 330 335
Asp Ser Phe Gly Trp Ala Leu Phe Ala Leu Phe Arg Leu Met Ala Gln
340 345 350
Asp Tyr Pro Glu Val Leu Tyr His Gln Ile Leu Tyr Ala Ser Gly Lys
355 360 365
Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser Phe Tyr
370 375 380
Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr Glu Glu Glu
385 390 395 400
Lys Gln Arg Val Gly Glu Ile Ser Lys Lys Ile Glu Pro Lys Phe Gln
405 410 415
Gln Thr Gly Lys Glu Leu Gln Glu Gly Asn Glu Thr Asp Glu Ala Lys
420 425 430
Thr Ile Gln Ile Glu Met Lys Lys Arg Ser Pro Ile Ser Thr Asp Thr
435 440 445
Ser Leu Asp Val Leu Glu Asp Ala Thr Leu Arg His Lys Glu Glu Leu
450 455 460
Glu Lys Ser Lys Lys Ile Cys Pro Leu Tyr Trp Tyr Lys Phe Ala Lys
465 470 475 480
Thr Phe Leu Ile Trp Asn Cys Ser Pro Cys Trp Leu Lys Leu Lys Glu
485 490 495
Phe Val His Arg Ile Ile Met Ala Pro Phe Thr Asp Leu Phe Leu Ile
500 505 510
Ile Cys Ile Ile Leu Asn Val Cys Phe Leu Thr Leu Glu His Tyr Pro
515 520 525
Met Ser Lys Gln Thr Asn Thr Leu Leu Asn Ile Gly Asn Leu Val Phe
530 535 540
Ile Gly Ile Phe Thr Ala Glu Met Ile Phe Lys Ile Ile Ala Met His
545 550 555 560
Pro Tyr Gly Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Met Ile
565 570 575
Val Phe His Gly Leu Ile Glu Leu Cys Leu Ala Asn Val Ala Gly Met
580 585 590
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
595 600 605
Tyr Trp Pro Thr Phe Gln Ile Leu Met Trp Ser Leu Ser Asn Ser Trp
610 615 620
Val Ala Leu Lys Asp Leu Val Leu Leu Leu Phe Thr Phe Ile Phe Phe
625 630 635 640
Ser Ala Ala Phe Gly Met Lys Leu Phe Gly Lys Asn Tyr Glu Glu Phe
645 650 655
Val Cys His Ile Asp Lys Asp Cys Gln Leu Pro Arg Trp His Met His
660 665 670
Asp Phe Phe His Ser Phe Leu Asn Val Phe Arg Ile Leu Cys Gly Glu
675 680 685
Trp Val Glu Thr Leu Trp Asp Cys Met Glu Val Ala Gly Gln Ser Trp
690 695 700
Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn Leu Leu Val
705 710 715 720
Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe Ser Ser Cys Lys Asp
725 730 735
Val Thr Ala Glu Glu Asn Asn Glu Ala Lys Asn Leu Gln Leu Ala Val
740 745 750
Ala Arg Ile Lys Lys Gly Ile Asn Tyr Val Leu Leu Lys Ile Leu Cys
755 760 765
Lys Thr Gln Asn Val Pro Lys Asp Thr Met Asp His Val Asn Glu Val
770 775 780
Tyr Val Lys Glu Asp Ile Ser Asp His Thr Leu Ser Glu Leu Ser Asn
785 790 795 800
Thr Gln Asp Phe Leu Lys Asp Lys Glu Lys Ser Ser Gly Thr Glu Lys
805 810 815
Asn Ala Thr Glu Asn Glu Ser Gln Ser Leu Ile Pro Ser Pro Ser Val
820 825 830
Ser Glu Thr Val Pro Ile Ala Ser Gly Glu Ser Asp Ile Glu Asn Leu
835 840 845
Asp Asn Lys Glu Ile Gln Ser Lys Ser Gly Asp Gly Gly Ser Lys Glu
850 855 860
Lys Ile Lys Gln Ser Ser Ser Ser Glu Cys Ser Thr Val Asp Ile Ala
865 870 875 880
Ile Ser Glu Glu Glu Glu Met Phe Tyr Gly Gly Glu Arg Ser Lys His
885 890 895
Leu Lys Asn Gly Cys Arg Arg Gly Ser Ser Leu Gly Gln Ile Ser Gly
900 905 910
Ala Ser Lys Lys Gly Lys Ile Trp Gln Asn Ile Arg Lys Thr Cys Cys
915 920 925
Lys Ile Val Glu Asn Asn Trp Phe Lys Cys Phe Ile Gly Leu Val Thr
930 935 940
Leu Leu Ser Thr Gly Thr Leu Ala Phe Glu Asp Ile Tyr Met Asp Gln
945 950 955 960
Arg Lys Thr Ile Lys Ile Leu Leu Glu Tyr Ala Asp Met Ile Phe Thr
965 970 975
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Met Ala Tyr Gly Phe
980 985 990
Lys Ala Tyr Phe Ser Asn Gly Trp Tyr Arg Leu Asp Phe Val Val Val
995 1000 1005
Ile Val Phe Cys Leu Ser Leu Ile Gly Lys Thr Arg Glu Glu Leu
1010 1015 1020
Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu
1025 1030 1035
Ser Gln Phe Glu Arg Met Lys Val Val Val Arg Ala Leu Ile Lys
1040 1045 1050
Thr Thr Leu Pro Thr Leu Asn Val Phe Leu Val Cys Leu Met Ile
1055 1060 1065
Trp Leu Ile Phe Ser Ile Met Gly Val Asp Leu Phe Ala Gly Arg
1070 1075 1080
Phe Tyr Glu Cys Ile Asp Pro Thr Ser Gly Glu Arg Phe Pro Ser
1085 1090 1095
Ser Glu Val Met Asn Lys Ser Arg Cys Glu Ser Leu Leu Phe Asn
1100 1105 1110
Glu Ser Met Leu Trp Glu Asn Ala Lys Met Asn Phe Asp Asn Val
1115 1120 1125
Gly Asn Gly Phe Leu Ser Leu Leu Gln Val Ala Thr Phe Asn Gly
1130 1135 1140
Trp Ile Thr Ile Met Asn Ser Ala Ile Asp Ser Val Ala Val Asn
1145 1150 1155
Ile Gln Pro His Phe Glu Val Asn Ile Tyr Met Tyr Cys Tyr Phe
1160 1165 1170
Ile Asn Phe Ile Ile Phe Gly Val Phe Leu Pro Leu Ser Met Leu
1175 1180 1185
Ile Thr Val Ile Ile Asp Asn Phe Asn Lys His Lys Ile Lys Leu
1190 1195 1200
Gly Gly Ser Asn Ile Phe Ile Thr Val Lys Gln Arg Lys Gln Tyr
1205 1210 1215
Arg Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser Gln Arg Pro Val
1220 1225 1230
Pro Arg Pro Leu Asn Lys Leu Gln Gly Phe Ile Phe Asp Val Val
1235 1240 1245
Thr Ser Gln Ala Phe Asn Val Ile Val Met Val Leu Ile Cys Phe
1250 1255 1260
Gln Ala Ile Ala Met Met Ile Asp Thr Asp Val Gln Ser Leu Gln
1265 1270 1275
Met Ser Ile Ala Leu Tyr Trp Ile Asn Ser Ile Phe Val Met Leu
1280 1285 1290
Tyr Thr Met Glu Cys Ile Leu Lys Leu Ile Ala Phe Arg Cys Phe
1295 1300 1305
Tyr Phe Thr Ile Ala Trp Asn Ile Phe Asp Phe Met Val Val Ile
1310 1315 1320
Phe Ser Ile Thr Gly Leu Cys Leu Pro Met Thr Val Gly Ser Tyr
1325 1330 1335
Leu Val Pro Pro Ser Leu Val Gln Leu Ile Leu Leu Ser Arg Ile
1340 1345 1350
Ile His Met Leu Arg Leu Gly Lys Gly Pro Lys Val Phe His Asn
1355 1360 1365
Leu Met Leu Pro Leu Met Leu Ser Leu Pro Ala Leu Leu Asn Ile
1370 1375 1380
Ile Leu Leu Ile Phe Leu Val Met Phe Ile Tyr Ala Val Phe Gly
1385 1390 1395
Met Tyr Asn Phe Ala Tyr Val Lys Lys Glu Ala Gly Ile Asn Asp
1400 1405 1410
Val Ser Asn Phe Glu Thr Phe Gly Asn Ser Met Leu Cys Leu Phe
1415 1420 1425
Gln Val Ala Ile Phe Ala Gly Trp Asp Gly Met Leu Asp Ala Ile
1430 1435 1440
Phe Asn Ser Lys Trp Ser Asp Cys Asp Pro Asp Lys Ile Asn Pro
1445 1450 1455
Gly Thr Gln Val Arg Gly Asp Cys Gly Asn Pro Ser Val Gly Ile
1460 1465 1470
Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp Leu Ile Ile Val
1475 1480 1485
Asn Met Tyr Ile Val Val Val Met Glu Phe Leu Asn Ile Ala Ser
1490 1495 1500
Lys Lys Lys Asn Lys Thr Leu Ser Glu Asp Asp Phe Arg Lys Phe
1505 1510 1515
Phe Gln Val Trp Lys Arg Phe Asp Pro Asp Arg Thr Gln Tyr Ile
1520 1525 1530
Asp Ser Ser Lys Leu Ser Asp Phe Ala Ala Ala Leu Asp Pro Pro
1535 1540 1545
Leu Phe Met Ala Lys Pro Asn Lys Gly Gln Leu Ile Ala Leu Asp
1550 1555 1560
Leu Pro Met Ala Val Gly Asp Arg Ile His Cys Leu Asp Ile Leu
1565 1570 1575
Leu Ala Phe Thr Lys Arg Val Met Gly Gln Asp Val Arg Met Glu
1580 1585 1590
Lys Val Val Ser Glu Ile Glu Ser Gly Phe Leu Leu Ala Asn Pro
1595 1600 1605
Phe Lys Ile Thr Cys Glu Pro Ile Thr Thr Thr Leu Lys Arg Lys
1610 1615 1620
Gln Glu Ala Val Ser Ala Thr Ile Ile Gln Arg Ala Tyr Lys Asn
1625 1630 1635
Tyr Arg Leu Arg Arg Asn Asp Lys Asn Thr Ser Asp Ile His Met
1640 1645 1650
Ile Asp Gly Asp Arg Asp Val His Ala Thr Lys Glu Gly Ala Tyr
1655 1660 1665
Phe Asp Lys Ala Lys Glu Lys Ser Pro Ile Gln Ser Gln Ile
1670 1675 1680
<210> 28
<211> 2009
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(2009)
<223> 人 Nav 1.1（图 9）
<400> 28
Met Glu Gln Thr Val Leu Val Pro Pro Gly Pro Asp Ser Phe Asn Phe
1 5 10 15
Phe Thr Arg Glu Ser Leu Ala Ala Ile Glu Arg Arg Ile Ala Glu Glu
20 25 30
Lys Ala Lys Asn Pro Lys Pro Asp Lys Lys Asp Asp Asp Glu Asn Gly
35 40 45
Pro Lys Pro Asn Ser Asp Leu Glu Ala Gly Lys Asn Leu Pro Phe Ile
50 55 60
Tyr Gly Asp Ile Pro Pro Glu Met Val Ser Glu Pro Leu Glu Asp Leu
65 70 75 80
Asp Pro Tyr Tyr Ile Asn Lys Lys Thr Phe Ile Val Leu Asn Lys Gly
85 90 95
Lys Ala Ile Phe Arg Phe Ser Ala Thr Ser Ala Leu Tyr Ile Leu Thr
100 105 110
Pro Phe Asn Pro Leu Arg Lys Ile Ala Ile Lys Ile Leu Val His Ser
115 120 125
Leu Phe Ser Met Leu Ile Met Cys Thr Ile Leu Thr Asn Cys Val Phe
130 135 140
Met Thr Met Ser Asn Pro Pro Asp Trp Thr Lys Asn Val Glu Tyr Thr
145 150 155 160
Phe Thr Gly Ile Tyr Thr Phe Glu Ser Leu Ile Lys Ile Ile Ala Arg
165 170 175
Gly Phe Cys Leu Glu Asp Phe Thr Phe Leu Arg Asp Pro Trp Asn Trp
180 185 190
Leu Asp Phe Thr Val Ile Thr Phe Ala Tyr Val Thr Glu Phe Val Asp
195 200 205
Leu Gly Asn Val Ser Ala Leu Arg Thr Phe Arg Val Leu Arg Ala Leu
210 215 220
Lys Thr Ile Ser Val Ile Pro Gly Leu Lys Thr Ile Val Gly Ala Leu
225 230 235 240
Ile Gln Ser Val Lys Lys Leu Ser Asp Val Met Ile Leu Thr Val Phe
245 250 255
Cys Leu Ser Val Phe Ala Leu Ile Gly Leu Gln Leu Phe Met Gly Asn
260 265 270
Leu Arg Asn Lys Cys Ile Gln Trp Pro Pro Thr Asn Ala Ser Leu Glu
275 280 285
Glu His Ser Ile Glu Lys Asn Ile Thr Val Asn Tyr Asn Gly Thr Leu
290 295 300
Ile Asn Glu Thr Val Phe Glu Phe Asp Trp Lys Ser Tyr Ile Gln Asp
305 310 315 320
Ser Arg Tyr His Tyr Phe Leu Glu Gly Phe Leu Asp Ala Leu Leu Cys
325 330 335
Gly Asn Ser Ser Asp Ala Gly Gln Cys Pro Glu Gly Tyr Met Cys Val
340 345 350
Lys Ala Gly Arg Asn Pro Asn Tyr Gly Tyr Thr Ser Phe Asp Thr Phe
355 360 365
Ser Trp Ala Phe Leu Ser Leu Phe Arg Leu Met Thr Gln Asp Phe Trp
370 375 380
Glu Asn Leu Tyr Gln Leu Thr Leu Arg Ala Ala Gly Lys Thr Tyr Met
385 390 395 400
Ile Phe Phe Val Leu Val Ile Phe Leu Gly Ser Phe Tyr Leu Ile Asn
405 410 415
Leu Ile Leu Ala Val Val Ala Met Ala Tyr Glu Glu Gln Asn Gln Ala
420 425 430
Thr Leu Glu Glu Ala Glu Gln Lys Glu Ala Glu Phe Gln Gln Met Ile
435 440 445
Glu Gln Leu Lys Lys Gln Gln Glu Ala Ala Gln Gln Ala Ala Thr Ala
450 455 460
Thr Ala Ser Glu His Ser Arg Glu Pro Ser Ala Ala Gly Arg Leu Ser
465 470 475 480
Asp Ser Ser Ser Glu Ala Ser Lys Leu Ser Ser Lys Ser Ala Lys Glu
485 490 495
Arg Arg Asn Arg Arg Lys Lys Arg Lys Gln Lys Glu Gln Ser Gly Gly
500 505 510
Glu Glu Lys Asp Glu Asp Glu Phe Gln Lys Ser Glu Ser Glu Asp Ser
515 520 525
Ile Arg Arg Lys Gly Phe Arg Phe Ser Ile Glu Gly Asn Arg Leu Thr
530 535 540
Tyr Glu Lys Arg Tyr Ser Ser Pro His Gln Ser Leu Leu Ser Ile Arg
545 550 555 560
Gly Ser Leu Phe Ser Pro Arg Arg Asn Ser Arg Thr Ser Leu Phe Ser
565 570 575
Phe Arg Gly Arg Ala Lys Asp Val Gly Ser Glu Asn Asp Phe Ala Asp
580 585 590
Asp Glu His Ser Thr Phe Glu Asp Asn Glu Ser Arg Arg Asp Ser Leu
595 600 605
Phe Val Pro Arg Arg His Gly Glu Arg Arg Asn Ser Asn Leu Ser Gln
610 615 620
Thr Ser Arg Ser Ser Arg Met Leu Ala Val Phe Pro Ala Asn Gly Lys
625 630 635 640
Met His Ser Thr Val Asp Cys Asn Gly Val Val Ser Leu Val Gly Gly
645 650 655
Pro Ser Val Pro Thr Ser Pro Val Gly Gln Leu Leu Pro Glu Val Ile
660 665 670
Ile Asp Lys Pro Ala Thr Asp Asp Asn Gly Thr Thr Thr Glu Thr Glu
675 680 685
Met Arg Lys Arg Arg Ser Ser Ser Phe His Val Ser Met Asp Phe Leu
690 695 700
Glu Asp Pro Ser Gln Arg Gln Arg Ala Met Ser Ile Ala Ser Ile Leu
705 710 715 720
Thr Asn Thr Val Glu Glu Leu Glu Glu Ser Arg Gln Lys Cys Pro Pro
725 730 735
Cys Trp Tyr Lys Phe Ser Asn Ile Phe Leu Ile Trp Asp Cys Ser Pro
740 745 750
Tyr Trp Leu Lys Val Lys His Val Val Asn Leu Val Val Met Asp Pro
755 760 765
Phe Val Asp Leu Ala Ile Thr Ile Cys Ile Val Leu Asn Thr Leu Phe
770 775 780
Met Ala Met Glu His Tyr Pro Met Thr Asp His Phe Asn Asn Val Leu
785 790 795 800
Thr Val Gly Asn Leu Val Phe Thr Gly Ile Phe Thr Ala Glu Met Phe
805 810 815
Leu Lys Ile Ile Ala Met Asp Pro Tyr Tyr Tyr Phe Gln Glu Gly Trp
820 825 830
Asn Ile Phe Asp Gly Phe Ile Val Thr Leu Ser Leu Val Glu Leu Gly
835 840 845
Leu Ala Asn Val Glu Gly Leu Ser Val Leu Arg Ser Phe Arg Leu Leu
850 855 860
Arg Val Phe Lys Leu Ala Lys Ser Trp Pro Thr Leu Asn Met Leu Ile
865 870 875 880
Lys Ile Ile Gly Asn Ser Val Gly Ala Leu Gly Asn Leu Thr Leu Val
885 890 895
Leu Ala Ile Ile Val Phe Ile Phe Ala Val Val Gly Met Gln Leu Phe
900 905 910
Gly Lys Ser Tyr Lys Asp Cys Val Cys Lys Ile Ala Ser Asp Cys Gln
915 920 925
Leu Pro Arg Trp His Met Asn Asp Phe Phe His Ser Phe Leu Ile Val
930 935 940
Phe Arg Val Leu Cys Gly Glu Trp Ile Glu Thr Met Trp Asp Cys Met
945 950 955 960
Glu Val Ala Gly Gln Ala Met Cys Leu Thr Val Phe Met Met Val Met
965 970 975
Val Ile Gly Asn Leu Val Val Leu Asn Leu Phe Leu Ala Leu Leu Leu
980 985 990
Ser Ser Phe Ser Ala Asp Asn Leu Ala Ala Thr Asp Asp Asp Asn Glu
995 1000 1005
Met Asn Asn Leu Gln Ile Ala Val Asp Arg Met His Lys Gly Val
1010 1015 1020
Ala Tyr Val Lys Arg Lys Ile Tyr Glu Phe Ile Gln Gln Ser Phe
1025 1030 1035
Ile Arg Lys Gln Lys Ile Leu Asp Glu Ile Lys Pro Leu Asp Asp
1040 1045 1050
Leu Asn Asn Lys Lys Asp Ser Cys Met Ser Asn His Thr Ala Glu
1055 1060 1065
Ile Gly Lys Asp Leu Asp Tyr Leu Lys Asp Val Asn Gly Thr Thr
1070 1075 1080
Ser Gly Ile Gly Thr Gly Ser Ser Val Glu Lys Tyr Ile Ile Asp
1085 1090 1095
Glu Ser Asp Tyr Met Ser Phe Ile Asn Asn Pro Ser Leu Thr Val
1100 1105 1110
Thr Val Pro Ile Ala Val Gly Glu Ser Asp Phe Glu Asn Leu Asn
1115 1120 1125
Thr Glu Asp Phe Ser Ser Glu Ser Asp Leu Glu Glu Ser Lys Glu
1130 1135 1140
Lys Leu Asn Glu Ser Ser Ser Ser Ser Glu Gly Ser Thr Val Asp
1145 1150 1155
Ile Gly Ala Pro Val Glu Glu Gln Pro Val Val Glu Pro Glu Glu
1160 1165 1170
Thr Leu Glu Pro Glu Ala Cys Phe Thr Glu Gly Cys Val Gln Arg
1175 1180 1185
Phe Lys Cys Cys Gln Ile Asn Val Glu Glu Gly Arg Gly Lys Gln
1190 1195 1200
Trp Trp Asn Leu Arg Arg Thr Cys Phe Arg Ile Val Glu His Asn
1205 1210 1215
Trp Phe Glu Thr Phe Ile Val Phe Met Ile Leu Leu Ser Ser Gly
1220 1225 1230
Ala Leu Ala Phe Glu Asp Ile Tyr Ile Asp Gln Arg Lys Thr Ile
1235 1240 1245
Lys Thr Met Leu Glu Tyr Ala Asp Lys Val Phe Thr Tyr Ile Phe
1250 1255 1260
Ile Leu Glu Met Leu Leu Lys Trp Val Ala Tyr Gly Tyr Gln Thr
1265 1270 1275
Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val Asp
1280 1285 1290
Val Ser Leu Val Ser Leu Thr Ala Asn Ala Leu Gly Tyr Ser Glu
1295 1300 1305
Leu Gly Ala Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg Pro
1310 1315 1320
Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val Asn
1325 1330 1335
Ala Leu Leu Gly Ala Ile Pro Ser Ile Met Asn Val Leu Leu Val
1340 1345 1350
Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile Met Gly Val Asn Leu
1355 1360 1365
Phe Ala Gly Lys Phe Tyr His Cys Ile Asn Thr Thr Thr Gly Asp
1370 1375 1380
Arg Phe Asp Ile Glu Asp Val Asn Asn His Thr Asp Cys Leu Lys
1385 1390 1395
Leu Ile Glu Arg Asn Glu Thr Ala Arg Trp Lys Asn Val Lys Val
1400 1405 1410
Asn Phe Asp Asn Val Gly Phe Gly Tyr Leu Ser Leu Leu Gln Val
1415 1420 1425
Ala Thr Phe Lys Gly Trp Met Asp Ile Met Tyr Ala Ala Val Asp
1430 1435 1440
Ser Arg Asn Val Glu Leu Gln Pro Lys Tyr Glu Glu Ser Leu Tyr
1445 1450 1455
Met Tyr Leu Tyr Phe Val Ile Phe Ile Ile Phe Gly Ser Phe Phe
1460 1465 1470
Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe Asn Gln
1475 1480 1485
Gln Lys Lys Lys Phe Gly Gly Gln Asp Ile Phe Met Thr Glu Glu
1490 1495 1500
Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys Lys
1505 1510 1515
Pro Gln Lys Pro Ile Pro Arg Pro Gly Asn Lys Phe Gln Gly Met
1520 1525 1530
Val Phe Asp Phe Val Thr Arg Gln Val Phe Asp Ile Ser Ile Met
1535 1540 1545
Ile Leu Ile Cys Leu Asn Met Val Thr Met Met Val Glu Thr Asp
1550 1555 1560
Asp Gln Ser Glu Tyr Val Thr Thr Ile Leu Ser Arg Ile Asn Leu
1565 1570 1575
Val Phe Ile Val Leu Phe Thr Gly Glu Cys Val Leu Lys Leu Ile
1580 1585 1590
Ser Leu Arg His Tyr Tyr Phe Thr Ile Gly Trp Asn Ile Phe Asp
1595 1600 1605
Phe Val Val Val Ile Leu Ser Ile Val Gly Met Phe Leu Ala Glu
1610 1615 1620
Leu Ile Glu Lys Tyr Phe Val Ser Pro Thr Leu Phe Arg Val Ile
1625 1630 1635
Arg Leu Ala Arg Ile Gly Arg Ile Leu Arg Leu Ile Lys Gly Ala
1640 1645 1650
Lys Gly Ile Arg Thr Leu Leu Phe Ala Leu Met Met Ser Leu Pro
1655 1660 1665
Ala Leu Phe Asn Ile Gly Leu Leu Leu Phe Leu Val Met Phe Ile
1670 1675 1680
Tyr Ala Ile Phe Gly Met Ser Asn Phe Ala Tyr Val Lys Arg Glu
1685 1690 1695
Val Gly Ile Asp Asp Met Phe Asn Phe Glu Thr Phe Gly Asn Ser
1700 1705 1710
Met Ile Cys Leu Phe Gln Ile Thr Thr Ser Ala Gly Trp Asp Gly
1715 1720 1725
Leu Leu Ala Pro Ile Leu Asn Ser Lys Pro Pro Asp Cys Asp Pro
1730 1735 1740
Asn Lys Val Asn Pro Gly Ser Ser Val Lys Gly Asp Cys Gly Asn
1745 1750 1755
Pro Ser Val Gly Ile Phe Phe Phe Val Ser Tyr Ile Ile Ile Ser
1760 1765 1770
Phe Leu Val Val Val Asn Met Tyr Ile Ala Val Ile Leu Glu Asn
1775 1780 1785
Phe Ser Val Ala Thr Glu Glu Ser Ala Glu Pro Leu Ser Glu Asp
1790 1795 1800
Asp Phe Glu Met Phe Tyr Glu Val Trp Glu Lys Phe Asp Pro Asp
1805 1810 1815
Ala Thr Gln Phe Met Glu Phe Glu Lys Leu Ser Gln Phe Ala Ala
1820 1825 1830
Ala Leu Glu Pro Pro Leu Asn Leu Pro Gln Pro Asn Lys Leu Gln
1835 1840 1845
Leu Ile Ala Met Asp Leu Pro Met Val Ser Gly Asp Arg Ile His
1850 1855 1860
Cys Leu Asp Ile Leu Phe Ala Phe Thr Lys Arg Val Leu Gly Glu
1865 1870 1875
Ser Gly Glu Met Asp Ala Leu Arg Ile Gln Met Glu Glu Arg Phe
1880 1885 1890
Met Ala Ser Asn Pro Ser Lys Val Ser Tyr Gln Pro Ile Thr Thr
1895 1900 1905
Thr Leu Lys Arg Lys Gln Glu Glu Val Ser Ala Val Ile Ile Gln
1910 1915 1920
Arg Ala Tyr Arg Arg His Leu Leu Lys Arg Thr Val Lys Gln Ala
1925 1930 1935
Ser Phe Thr Tyr Asn Lys Asn Lys Ile Lys Gly Gly Ala Asn Leu
1940 1945 1950
Leu Ile Lys Glu Asp Met Ile Ile Asp Arg Ile Asn Glu Asn Ser
1955 1960 1965
Ile Thr Glu Lys Thr Asp Leu Thr Met Ser Thr Ala Ala Cys Pro
1970 1975 1980
Pro Ser Tyr Asp Arg Val Thr Lys Pro Ile Val Glu Lys His Glu
1985 1990 1995
Gln Glu Gly Lys Asp Glu Lys Ala Lys Gly Lys
2000 2005
<210> 29
<211> 2005
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(2005)
<223> 人 Nav 1.2（图 9）
<400> 29
Met Ala Gln Ser Val Leu Val Pro Pro Gly Pro Asp Ser Phe Arg Phe
1 5 10 15
Phe Thr Arg Glu Ser Leu Ala Ala Ile Glu Gln Arg Ile Ala Glu Glu
20 25 30
Lys Ala Lys Arg Pro Lys Gln Glu Arg Lys Asp Glu Asp Asp Glu Asn
35 40 45
Gly Pro Lys Pro Asn Ser Asp Leu Glu Ala Gly Lys Ser Leu Pro Phe
50 55 60
Ile Tyr Gly Asp Ile Pro Pro Glu Met Val Ser Val Pro Leu Glu Asp
65 70 75 80
Leu Asp Pro Tyr Tyr Ile Asn Lys Lys Thr Phe Ile Val Leu Asn Lys
85 90 95
Gly Lys Ala Ile Ser Arg Phe Ser Ala Thr Pro Ala Leu Tyr Ile Leu
100 105 110
Thr Pro Phe Asn Pro Ile Arg Lys Leu Ala Ile Lys Ile Leu Val His
115 120 125
Ser Leu Phe Asn Met Leu Ile Met Cys Thr Ile Leu Thr Asn Cys Val
130 135 140
Phe Met Thr Met Ser Asn Pro Pro Asp Trp Thr Lys Asn Val Glu Tyr
145 150 155 160
Thr Phe Thr Gly Ile Tyr Thr Phe Glu Ser Leu Ile Lys Ile Leu Ala
165 170 175
Arg Gly Phe Cys Leu Glu Asp Phe Thr Phe Leu Arg Asp Pro Trp Asn
180 185 190
Trp Leu Asp Phe Thr Val Ile Thr Phe Ala Tyr Val Thr Glu Phe Val
195 200 205
Asp Leu Gly Asn Val Ser Ala Leu Arg Thr Phe Arg Val Leu Arg Ala
210 215 220
Leu Lys Thr Ile Ser Val Ile Pro Gly Leu Lys Thr Ile Val Gly Ala
225 230 235 240
Leu Ile Gln Ser Val Lys Lys Leu Ser Asp Val Met Ile Leu Thr Val
245 250 255
Phe Cys Leu Ser Val Phe Ala Leu Ile Gly Leu Gln Leu Phe Met Gly
260 265 270
Asn Leu Arg Asn Lys Cys Leu Gln Trp Pro Pro Asp Asn Ser Ser Phe
275 280 285
Glu Ile Asn Ile Thr Ser Phe Phe Asn Asn Ser Leu Asp Gly Asn Gly
290 295 300
Thr Thr Phe Asn Arg Thr Val Ser Ile Phe Asn Trp Asp Glu Tyr Ile
305 310 315 320
Glu Asp Lys Ser His Phe Tyr Phe Leu Glu Gly Gln Asn Asp Ala Leu
325 330 335
Leu Cys Gly Asn Ser Ser Asp Ala Gly Gln Cys Pro Glu Gly Tyr Ile
340 345 350
Cys Val Lys Ala Gly Arg Asn Pro Asn Tyr Gly Tyr Thr Ser Phe Asp
355 360 365
Thr Phe Ser Trp Ala Phe Leu Ser Leu Phe Arg Leu Met Thr Gln Asp
370 375 380
Phe Trp Glu Asn Leu Tyr Gln Leu Thr Leu Arg Ala Ala Gly Lys Thr
385 390 395 400
Tyr Met Ile Phe Phe Val Leu Val Ile Phe Leu Gly Ser Phe Tyr Leu
405 410 415
Ile Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr Glu Glu Gln Asn
420 425 430
Gln Ala Thr Leu Glu Glu Ala Glu Gln Lys Glu Ala Glu Phe Gln Gln
435 440 445
Met Leu Glu Gln Leu Lys Lys Gln Gln Glu Glu Ala Gln Ala Ala Ala
450 455 460
Ala Ala Ala Ser Ala Glu Ser Arg Asp Phe Ser Gly Ala Gly Gly Ile
465 470 475 480
Gly Val Phe Ser Glu Ser Ser Ser Val Ala Ser Lys Leu Ser Ser Lys
485 490 495
Ser Glu Lys Glu Leu Lys Asn Arg Arg Lys Lys Lys Lys Gln Lys Glu
500 505 510
Gln Ser Gly Glu Glu Glu Lys Asn Asp Arg Val Arg Lys Ser Glu Ser
515 520 525
Glu Asp Ser Ile Arg Arg Lys Gly Phe Arg Phe Ser Leu Glu Gly Ser
530 535 540
Arg Leu Thr Tyr Glu Lys Arg Phe Ser Ser Pro His Gln Ser Leu Leu
545 550 555 560
Ser Ile Arg Gly Ser Leu Phe Ser Pro Arg Arg Asn Ser Arg Ala Ser
565 570 575
Leu Phe Ser Phe Arg Gly Arg Ala Lys Asp Ile Gly Ser Glu Asn Asp
580 585 590
Phe Ala Asp Asp Glu His Ser Thr Phe Glu Asp Asn Asp Ser Arg Arg
595 600 605
Asp Ser Leu Phe Val Pro His Arg His Gly Glu Arg Arg His Ser Asn
610 615 620
Val Ser Gln Ala Ser Arg Ala Ser Arg Val Leu Pro Ile Leu Pro Met
625 630 635 640
Asn Gly Lys Met His Ser Ala Val Asp Cys Asn Gly Val Val Ser Leu
645 650 655
Val Gly Gly Pro Ser Thr Leu Thr Ser Ala Gly Gln Leu Leu Pro Glu
660 665 670
Gly Thr Thr Thr Glu Thr Glu Ile Arg Lys Arg Arg Ser Ser Ser Tyr
675 680 685
His Val Ser Met Asp Leu Leu Glu Asp Pro Thr Ser Arg Gln Arg Ala
690 695 700
Met Ser Ile Ala Ser Ile Leu Thr Asn Thr Met Glu Glu Leu Glu Glu
705 710 715 720
Ser Arg Gln Lys Cys Pro Pro Cys Trp Tyr Lys Phe Ala Asn Met Cys
725 730 735
Leu Ile Trp Asp Cys Cys Lys Pro Trp Leu Lys Val Lys His Leu Val
740 745 750
Asn Leu Val Val Met Asp Pro Phe Val Asp Leu Ala Ile Thr Ile Cys
755 760 765
Ile Val Leu Asn Thr Leu Phe Met Ala Met Glu His Tyr Pro Met Thr
770 775 780
Glu Gln Phe Ser Ser Val Leu Ser Val Gly Asn Leu Val Phe Thr Gly
785 790 795 800
Ile Phe Thr Ala Glu Met Phe Leu Lys Ile Ile Ala Met Asp Pro Tyr
805 810 815
Tyr Tyr Phe Gln Glu Gly Trp Asn Ile Phe Asp Gly Phe Ile Val Ser
820 825 830
Leu Ser Leu Met Glu Leu Gly Leu Ala Asn Val Glu Gly Leu Ser Val
835 840 845
Leu Arg Ser Phe Arg Leu Leu Arg Val Phe Lys Leu Ala Lys Ser Trp
850 855 860
Pro Thr Leu Asn Met Leu Ile Lys Ile Ile Gly Asn Ser Val Gly Ala
865 870 875 880
Leu Gly Asn Leu Thr Leu Val Leu Ala Ile Ile Val Phe Ile Phe Ala
885 890 895
Val Val Gly Met Gln Leu Phe Gly Lys Ser Tyr Lys Glu Cys Val Cys
900 905 910
Lys Ile Ser Asn Asp Cys Glu Leu Pro Arg Trp His Met His Asp Phe
915 920 925
Phe His Ser Phe Leu Ile Val Phe Arg Val Leu Cys Gly Glu Trp Ile
930 935 940
Glu Thr Met Trp Asp Cys Met Glu Val Ala Gly Gln Thr Met Cys Leu
945 950 955 960
Thr Val Phe Met Met Val Met Val Ile Gly Asn Leu Val Val Leu Asn
965 970 975
Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe Ser Ser Asp Asn Leu Ala
980 985 990
Ala Thr Asp Asp Asp Asn Glu Met Asn Asn Leu Gln Ile Ala Val Gly
995 1000 1005
Arg Met Gln Lys Gly Ile Asp Phe Val Lys Arg Lys Ile Arg Glu
1010 1015 1020
Phe Ile Gln Lys Ala Phe Val Arg Lys Gln Lys Ala Leu Asp Glu
1025 1030 1035
Ile Lys Pro Leu Glu Asp Leu Asn Asn Lys Lys Asp Ser Cys Ile
1040 1045 1050
Ser Asn His Thr Thr Ile Glu Ile Gly Lys Asp Leu Asn Tyr Leu
1055 1060 1065
Lys Asp Gly Asn Gly Thr Thr Ser Gly Ile Gly Ser Ser Val Glu
1070 1075 1080
Lys Tyr Val Val Asp Glu Ser Asp Tyr Met Ser Phe Ile Asn Asn
1085 1090 1095
Pro Ser Leu Thr Val Thr Val Pro Ile Ala Val Gly Glu Ser Asp
1100 1105 1110
Phe Glu Asn Leu Asn Thr Glu Glu Phe Ser Ser Glu Ser Asp Met
1115 1120 1125
Glu Glu Ser Lys Glu Lys Leu Asn Ala Thr Ser Ser Ser Glu Gly
1130 1135 1140
Ser Thr Val Asp Ile Gly Ala Pro Ala Glu Gly Glu Gln Pro Glu
1145 1150 1155
Val Glu Pro Glu Glu Ser Leu Glu Pro Glu Ala Cys Phe Thr Glu
1160 1165 1170
Asp Cys Val Arg Lys Phe Lys Cys Cys Gln Ile Ser Ile Glu Glu
1175 1180 1185
Gly Lys Gly Lys Leu Trp Trp Asn Leu Arg Lys Thr Cys Tyr Lys
1190 1195 1200
Ile Val Glu His Asn Trp Phe Glu Thr Phe Ile Val Phe Met Ile
1205 1210 1215
Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu
1220 1225 1230
Gln Arg Lys Thr Ile Lys Thr Met Leu Glu Tyr Ala Asp Lys Val
1235 1240 1245
Phe Thr Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Val Ala
1250 1255 1260
Tyr Gly Phe Gln Val Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp
1265 1270 1275
Phe Leu Ile Val Asp Val Ser Leu Val Ser Leu Thr Ala Asn Ala
1280 1285 1290
Leu Gly Tyr Ser Glu Leu Gly Ala Ile Lys Ser Leu Arg Thr Leu
1295 1300 1305
Arg Ala Leu Arg Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met
1310 1315 1320
Arg Val Val Val Asn Ala Leu Leu Gly Ala Ile Pro Ser Ile Met
1325 1330 1335
Asn Val Leu Leu Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile
1340 1345 1350
Met Gly Val Asn Leu Phe Ala Gly Lys Phe Tyr His Cys Ile Asn
1355 1360 1365
Tyr Thr Thr Gly Glu Met Phe Asp Val Ser Val Val Asn Asn Tyr
1370 1375 1380
Ser Glu Cys Lys Ala Leu Ile Glu Ser Asn Gln Thr Ala Arg Trp
1385 1390 1395
Lys Asn Val Lys Val Asn Phe Asp Asn Val Gly Leu Gly Tyr Leu
1400 1405 1410
Ser Leu Leu Gln Val Ala Thr Phe Lys Gly Trp Met Asp Ile Met
1415 1420 1425
Tyr Ala Ala Val Asp Ser Arg Asn Val Glu Leu Gln Pro Lys Tyr
1430 1435 1440
Glu Asp Asn Leu Tyr Met Tyr Leu Tyr Phe Val Ile Phe Ile Ile
1445 1450 1455
Phe Gly Ser Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile
1460 1465 1470
Asp Asn Phe Asn Gln Gln Lys Lys Lys Phe Gly Gly Gln Asp Ile
1475 1480 1485
Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys
1490 1495 1500
Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg Pro Ala Asn
1505 1510 1515
Lys Phe Gln Gly Met Val Phe Asp Phe Val Thr Lys Gln Val Phe
1520 1525 1530
Asp Ile Ser Ile Met Ile Leu Ile Cys Leu Asn Met Val Thr Met
1535 1540 1545
Met Val Glu Thr Asp Asp Gln Ser Gln Glu Met Thr Asn Ile Leu
1550 1555 1560
Tyr Trp Ile Asn Leu Val Phe Ile Val Leu Phe Thr Gly Glu Cys
1565 1570 1575
Val Leu Lys Leu Ile Ser Leu Arg Tyr Tyr Tyr Phe Thr Ile Gly
1580 1585 1590
Trp Asn Ile Phe Asp Phe Val Val Val Ile Leu Ser Ile Val Gly
1595 1600 1605
Met Phe Leu Ala Glu Leu Ile Glu Lys Tyr Phe Val Ser Pro Thr
1610 1615 1620
Leu Phe Arg Val Ile Arg Leu Ala Arg Ile Gly Arg Ile Leu Arg
1625 1630 1635
Leu Ile Lys Gly Ala Lys Gly Ile Arg Thr Leu Leu Phe Ala Leu
1640 1645 1650
Met Met Ser Leu Pro Ala Leu Phe Asn Ile Gly Leu Leu Leu Phe
1655 1660 1665
Leu Val Met Phe Ile Tyr Ala Ile Phe Gly Met Ser Asn Phe Ala
1670 1675 1680
Tyr Val Lys Arg Glu Val Gly Ile Asp Asp Met Phe Asn Phe Glu
1685 1690 1695
Thr Phe Gly Asn Ser Met Ile Cys Leu Phe Gln Ile Thr Thr Ser
1700 1705 1710
Ala Gly Trp Asp Gly Leu Leu Ala Pro Ile Leu Asn Ser Gly Pro
1715 1720 1725
Pro Asp Cys Asp Pro Asp Lys Asp His Pro Gly Ser Ser Val Lys
1730 1735 1740
Gly Asp Cys Gly Asn Pro Ser Val Gly Ile Phe Phe Phe Val Ser
1745 1750 1755
Tyr Ile Ile Ile Ser Phe Leu Val Val Val Asn Met Tyr Ile Ala
1760 1765 1770
Val Ile Leu Glu Asn Phe Ser Val Ala Thr Glu Glu Ser Ala Glu
1775 1780 1785
Pro Leu Ser Glu Asp Asp Phe Glu Met Phe Tyr Glu Val Trp Glu
1790 1795 1800
Lys Phe Asp Pro Asp Ala Thr Gln Phe Ile Glu Phe Ala Lys Leu
1805 1810 1815
Ser Asp Phe Ala Asp Ala Leu Asp Pro Pro Leu Leu Ile Ala Lys
1820 1825 1830
Pro Asn Lys Val Gln Leu Ile Ala Met Asp Leu Pro Met Val Ser
1835 1840 1845
Gly Asp Arg Ile His Cys Leu Asp Ile Leu Phe Ala Phe Thr Lys
1850 1855 1860
Arg Val Leu Gly Glu Ser Gly Glu Met Asp Ala Leu Arg Ile Gln
1865 1870 1875
Met Glu Glu Arg Phe Met Ala Ser Asn Pro Ser Lys Val Ser Tyr
1880 1885 1890
Glu Pro Ile Thr Thr Thr Leu Lys Arg Lys Gln Glu Glu Val Ser
1895 1900 1905
Ala Ile Ile Ile Gln Arg Ala Tyr Arg Arg Tyr Leu Leu Lys Gln
1910 1915 1920
Lys Val Lys Lys Val Ser Ser Ile Tyr Lys Lys Asp Lys Gly Lys
1925 1930 1935
Glu Cys Asp Gly Thr Pro Ile Lys Glu Asp Thr Leu Ile Asp Lys
1940 1945 1950
Leu Asn Glu Asn Ser Thr Pro Glu Lys Thr Asp Met Thr Pro Ser
1955 1960 1965
Thr Thr Ser Pro Pro Ser Tyr Asp Ser Val Thr Lys Pro Glu Lys
1970 1975 1980
Glu Lys Phe Glu Lys Asp Lys Ser Glu Lys Glu Asp Lys Gly Lys
1985 1990 1995
Asp Ile Arg Glu Ser Lys Lys
2000 2005
<210> 30
<211> 2000
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(2000)
<223> 人 Nav 1.3（图 9）
<400> 30
Met Ala Gln Ala Leu Leu Val Pro Pro Gly Pro Glu Ser Phe Arg Leu
1 5 10 15
Phe Thr Arg Glu Ser Leu Ala Ala Ile Glu Lys Arg Ala Ala Glu Glu
20 25 30
Lys Ala Lys Lys Pro Lys Lys Glu Gln Asp Asn Asp Asp Glu Asn Lys
35 40 45
Pro Lys Pro Asn Ser Asp Leu Glu Ala Gly Lys Asn Leu Pro Phe Ile
50 55 60
Tyr Gly Asp Ile Pro Pro Glu Met Val Ser Glu Pro Leu Glu Asp Leu
65 70 75 80
Asp Pro Tyr Tyr Ile Asn Lys Lys Thr Phe Ile Val Met Asn Lys Gly
85 90 95
Lys Ala Ile Phe Arg Phe Ser Ala Thr Ser Ala Leu Tyr Ile Leu Thr
100 105 110
Pro Leu Asn Pro Val Arg Lys Ile Ala Ile Lys Ile Leu Val His Ser
115 120 125
Leu Phe Ser Met Leu Ile Met Cys Thr Ile Leu Thr Asn Cys Val Phe
130 135 140
Met Thr Leu Ser Asn Pro Pro Asp Trp Thr Lys Asn Val Glu Tyr Thr
145 150 155 160
Phe Thr Gly Ile Tyr Thr Phe Glu Ser Leu Ile Lys Ile Leu Ala Arg
165 170 175
Gly Phe Cys Leu Glu Asp Phe Thr Phe Leu Arg Asp Pro Trp Asn Trp
180 185 190
Leu Asp Phe Ser Val Ile Val Met Ala Tyr Val Thr Glu Phe Val Ser
195 200 205
Leu Gly Asn Val Ser Ala Leu Arg Thr Phe Arg Val Leu Arg Ala Leu
210 215 220
Lys Thr Ile Ser Val Ile Pro Gly Leu Lys Thr Ile Val Gly Ala Leu
225 230 235 240
Ile Gln Ser Val Lys Lys Leu Ser Asp Val Met Ile Leu Thr Val Phe
245 250 255
Cys Leu Ser Val Phe Ala Leu Ile Gly Leu Gln Leu Phe Met Gly Asn
260 265 270
Leu Arg Asn Lys Cys Leu Gln Trp Pro Pro Ser Asp Ser Ala Phe Glu
275 280 285
Thr Asn Thr Thr Ser Tyr Phe Asn Gly Thr Met Asp Ser Asn Gly Thr
290 295 300
Phe Val Asn Val Thr Met Ser Thr Phe Asn Trp Lys Asp Tyr Ile Gly
305 310 315 320
Asp Asp Ser His Phe Tyr Val Leu Asp Gly Gln Lys Asp Pro Leu Leu
325 330 335
Cys Gly Asn Gly Ser Asp Ala Gly Gln Cys Pro Glu Gly Tyr Ile Cys
340 345 350
Val Lys Ala Gly Arg Asn Pro Asn Tyr Gly Tyr Thr Ser Phe Asp Thr
355 360 365
Phe Ser Trp Ala Phe Leu Ser Leu Phe Arg Leu Met Thr Gln Asp Tyr
370 375 380
Trp Glu Asn Leu Tyr Gln Leu Thr Leu Arg Ala Ala Gly Lys Thr Tyr
385 390 395 400
Met Ile Phe Phe Val Leu Val Ile Phe Leu Gly Ser Phe Tyr Leu Val
405 410 415
Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr Glu Glu Gln Asn Gln
420 425 430
Ala Thr Leu Glu Glu Ala Glu Gln Lys Glu Ala Glu Phe Gln Gln Met
435 440 445
Leu Glu Gln Leu Lys Lys Gln Gln Glu Glu Ala Gln Ala Val Ala Ala
450 455 460
Ala Ser Ala Ala Ser Arg Asp Phe Ser Gly Ile Gly Gly Leu Gly Glu
465 470 475 480
Leu Leu Glu Ser Ser Ser Glu Ala Ser Lys Leu Ser Ser Lys Ser Ala
485 490 495
Lys Glu Trp Arg Asn Arg Arg Lys Lys Arg Arg Gln Arg Glu His Leu
500 505 510
Glu Gly Asn Asn Lys Gly Glu Arg Asp Ser Phe Pro Lys Ser Glu Ser
515 520 525
Glu Asp Ser Val Lys Arg Ser Ser Phe Leu Phe Ser Met Asp Gly Asn
530 535 540
Arg Leu Thr Ser Asp Lys Lys Phe Cys Ser Pro His Gln Ser Leu Leu
545 550 555 560
Ser Ile Arg Gly Ser Leu Phe Ser Pro Arg Arg Asn Ser Lys Thr Ser
565 570 575
Ile Phe Ser Phe Arg Gly Arg Ala Lys Asp Val Gly Ser Glu Asn Asp
580 585 590
Phe Ala Asp Asp Glu His Ser Thr Phe Glu Asp Ser Glu Ser Arg Arg
595 600 605
Asp Ser Leu Phe Val Pro His Arg His Gly Glu Arg Arg Asn Ser Asn
610 615 620
Val Ser Gln Ala Ser Met Ser Ser Arg Met Val Pro Gly Leu Pro Ala
625 630 635 640
Asn Gly Lys Met His Ser Thr Val Asp Cys Asn Gly Val Val Ser Leu
645 650 655
Val Gly Gly Pro Ser Ala Leu Thr Ser Pro Thr Gly Gln Leu Pro Pro
660 665 670
Glu Gly Thr Thr Thr Glu Thr Glu Val Arg Lys Arg Arg Leu Ser Ser
675 680 685
Tyr Gln Ile Ser Met Glu Met Leu Glu Asp Ser Ser Gly Arg Gln Arg
690 695 700
Ala Val Ser Ile Ala Ser Ile Leu Thr Asn Thr Met Glu Glu Leu Glu
705 710 715 720
Glu Ser Arg Gln Lys Cys Pro Pro Cys Trp Tyr Arg Phe Ala Asn Val
725 730 735
Phe Leu Ile Trp Asp Cys Cys Asp Ala Trp Leu Lys Val Lys His Leu
740 745 750
Val Asn Leu Ile Val Met Asp Pro Phe Val Asp Leu Ala Ile Thr Ile
755 760 765
Cys Ile Val Leu Asn Thr Leu Phe Met Ala Met Glu His Tyr Pro Met
770 775 780
Thr Glu Gln Phe Ser Ser Val Leu Thr Val Gly Asn Leu Val Phe Thr
785 790 795 800
Gly Ile Phe Thr Ala Glu Met Val Leu Lys Ile Ile Ala Met Asp Pro
805 810 815
Tyr Tyr Tyr Phe Gln Glu Gly Trp Asn Ile Phe Asp Gly Ile Ile Val
820 825 830
Ser Leu Ser Leu Met Glu Leu Gly Leu Ser Asn Val Glu Gly Leu Ser
835 840 845
Val Leu Arg Ser Phe Arg Leu Leu Arg Val Phe Lys Leu Ala Lys Ser
850 855 860
Trp Pro Thr Leu Asn Met Leu Ile Lys Ile Ile Gly Asn Ser Val Gly
865 870 875 880
Ala Leu Gly Asn Leu Thr Leu Val Leu Ala Ile Ile Val Phe Ile Phe
885 890 895
Ala Val Val Gly Met Gln Leu Phe Gly Lys Ser Tyr Lys Glu Cys Val
900 905 910
Cys Lys Ile Asn Asp Asp Cys Thr Leu Pro Arg Trp His Met Asn Asp
915 920 925
Phe Phe His Ser Phe Leu Ile Val Phe Arg Val Leu Cys Gly Glu Trp
930 935 940
Ile Glu Thr Met Trp Asp Cys Met Glu Val Ala Gly Gln Thr Met Cys
945 950 955 960
Leu Ile Val Phe Met Leu Val Met Val Ile Gly Asn Leu Val Val Leu
965 970 975
Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe Ser Ser Asp Asn Leu
980 985 990
Ala Ala Thr Asp Asp Asp Asn Glu Met Asn Asn Leu Gln Ile Ala Val
995 1000 1005
Gly Arg Met Gln Lys Gly Ile Asp Tyr Val Lys Asn Lys Met Arg
1010 1015 1020
Glu Cys Phe Gln Lys Ala Phe Phe Arg Lys Pro Lys Val Ile Glu
1025 1030 1035
Ile His Glu Gly Asn Lys Ile Asp Ser Cys Met Ser Asn Asn Thr
1040 1045 1050
Gly Ile Glu Ile Ser Lys Glu Leu Asn Tyr Leu Arg Asp Gly Asn
1055 1060 1065
Gly Thr Thr Ser Gly Val Gly Thr Gly Ser Ser Val Glu Lys Tyr
1070 1075 1080
Val Ile Asp Glu Asn Asp Tyr Met Ser Phe Ile Asn Asn Pro Ser
1085 1090 1095
Leu Thr Val Thr Val Pro Ile Ala Val Gly Glu Ser Asp Phe Glu
1100 1105 1110
Asn Leu Asn Thr Glu Glu Phe Ser Ser Glu Ser Glu Leu Glu Glu
1115 1120 1125
Ser Lys Glu Lys Leu Asn Ala Thr Ser Ser Ser Glu Gly Ser Thr
1130 1135 1140
Val Asp Val Val Leu Pro Arg Glu Gly Glu Gln Ala Glu Thr Glu
1145 1150 1155
Pro Glu Glu Asp Leu Lys Pro Glu Ala Cys Phe Thr Glu Gly Cys
1160 1165 1170
Ile Lys Lys Phe Pro Phe Cys Gln Val Ser Thr Glu Glu Gly Lys
1175 1180 1185
Gly Lys Ile Trp Trp Asn Leu Arg Lys Thr Cys Tyr Ser Ile Val
1190 1195 1200
Glu His Asn Trp Phe Glu Thr Phe Ile Val Phe Met Ile Leu Leu
1205 1210 1215
Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Gln Arg
1220 1225 1230
Lys Thr Ile Lys Thr Met Leu Glu Tyr Ala Asp Lys Val Phe Thr
1235 1240 1245
Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Val Ala Tyr Gly
1250 1255 1260
Phe Gln Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu
1265 1270 1275
Ile Val Asp Val Ser Leu Val Ser Leu Val Ala Asn Ala Leu Gly
1280 1285 1290
Tyr Ser Glu Leu Gly Ala Ile Lys Ser Leu Arg Thr Leu Arg Ala
1295 1300 1305
Leu Arg Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val
1310 1315 1320
Val Val Asn Ala Leu Val Gly Ala Ile Pro Ser Ile Met Asn Val
1325 1330 1335
Leu Leu Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile Met Gly
1340 1345 1350
Val Asn Leu Phe Ala Gly Lys Phe Tyr His Cys Val Asn Met Thr
1355 1360 1365
Thr Gly Asn Met Phe Asp Ile Ser Asp Val Asn Asn Leu Ser Asp
1370 1375 1380
Cys Gln Ala Leu Gly Lys Gln Ala Arg Trp Lys Asn Val Lys Val
1385 1390 1395
Asn Phe Asp Asn Val Gly Ala Gly Tyr Leu Ala Leu Leu Gln Val
1400 1405 1410
Ala Thr Phe Lys Gly Trp Met Asp Ile Met Tyr Ala Ala Val Asp
1415 1420 1425
Ser Arg Asp Val Lys Leu Gln Pro Val Tyr Glu Glu Asn Leu Tyr
1430 1435 1440
Met Tyr Leu Tyr Phe Val Ile Phe Ile Ile Phe Gly Ser Phe Phe
1445 1450 1455
Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe Asn Gln
1460 1465 1470
Gln Lys Lys Lys Phe Gly Gly Gln Asp Ile Phe Met Thr Glu Glu
1475 1480 1485
Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys Lys
1490 1495 1500
Pro Gln Lys Pro Ile Pro Arg Pro Ala Asn Lys Phe Gln Gly Met
1505 1510 1515
Val Phe Asp Phe Val Thr Arg Gln Val Phe Asp Ile Ser Ile Met
1520 1525 1530
Ile Leu Ile Cys Leu Asn Met Val Thr Met Met Val Glu Thr Asp
1535 1540 1545
Asp Gln Gly Lys Tyr Met Thr Leu Val Leu Ser Arg Ile Asn Leu
1550 1555 1560
Val Phe Ile Val Leu Phe Thr Gly Glu Phe Val Leu Lys Leu Val
1565 1570 1575
Ser Leu Arg His Tyr Tyr Phe Thr Ile Gly Trp Asn Ile Phe Asp
1580 1585 1590
Phe Val Val Val Ile Leu Ser Ile Val Gly Met Phe Leu Ala Glu
1595 1600 1605
Met Ile Glu Lys Tyr Phe Val Ser Pro Thr Leu Phe Arg Val Ile
1610 1615 1620
Arg Leu Ala Arg Ile Gly Arg Ile Leu Arg Leu Ile Lys Gly Ala
1625 1630 1635
Lys Gly Ile Arg Thr Leu Leu Phe Ala Leu Met Met Ser Leu Pro
1640 1645 1650
Ala Leu Phe Asn Ile Gly Leu Leu Leu Phe Leu Val Met Phe Ile
1655 1660 1665
Tyr Ala Ile Phe Gly Met Ser Asn Phe Ala Tyr Val Lys Lys Glu
1670 1675 1680
Ala Gly Ile Asp Asp Met Phe Asn Phe Glu Thr Phe Gly Asn Ser
1685 1690 1695
Met Ile Cys Leu Phe Gln Ile Thr Thr Ser Ala Gly Trp Asp Gly
1700 1705 1710
Leu Leu Ala Pro Ile Leu Asn Ser Ala Pro Pro Asp Cys Asp Pro
1715 1720 1725
Asp Thr Ile His Pro Gly Ser Ser Val Lys Gly Asp Cys Gly Asn
1730 1735 1740
Pro Ser Val Gly Ile Phe Phe Phe Val Ser Tyr Ile Ile Ile Ser
1745 1750 1755
Phe Leu Val Val Val Asn Met Tyr Ile Ala Val Ile Leu Glu Asn
1760 1765 1770
Phe Ser Val Ala Thr Glu Glu Ser Ala Glu Pro Leu Ser Glu Asp
1775 1780 1785
Asp Phe Glu Met Phe Tyr Glu Val Trp Glu Lys Phe Asp Pro Asp
1790 1795 1800
Ala Thr Gln Phe Ile Glu Phe Ser Lys Leu Ser Asp Phe Ala Ala
1805 1810 1815
Ala Leu Asp Pro Pro Leu Leu Ile Ala Lys Pro Asn Lys Val Gln
1820 1825 1830
Leu Ile Ala Met Asp Leu Pro Met Val Ser Gly Asp Arg Ile His
1835 1840 1845
Cys Leu Asp Ile Leu Phe Ala Phe Thr Lys Arg Val Leu Gly Glu
1850 1855 1860
Ser Gly Glu Met Asp Ala Leu Arg Ile Gln Met Glu Asp Arg Phe
1865 1870 1875
Met Ala Ser Asn Pro Ser Lys Val Ser Tyr Glu Pro Ile Thr Thr
1880 1885 1890
Thr Leu Lys Arg Lys Gln Glu Glu Val Ser Ala Ala Ile Ile Gln
1895 1900 1905
Arg Asn Phe Arg Cys Tyr Leu Leu Lys Gln Arg Leu Lys Asn Ile
1910 1915 1920
Ser Ser Asn Tyr Asn Lys Glu Ala Ile Lys Gly Arg Ile Asp Leu
1925 1930 1935
Pro Ile Lys Gln Asp Met Ile Ile Asp Lys Leu Asn Gly Asn Ser
1940 1945 1950
Thr Pro Glu Lys Thr Asp Gly Ser Ser Ser Thr Thr Ser Pro Pro
1955 1960 1965
Ser Tyr Asp Ser Val Thr Lys Pro Asp Lys Glu Lys Phe Glu Lys
1970 1975 1980
Asp Lys Pro Glu Lys Glu Ser Lys Gly Lys Glu Val Arg Glu Asn
1985 1990 1995
Gln Lys
2000
<210> 31
<211> 1788
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(1788)
<223> 人 Nav 1.4（图 9）
<400> 31
Met Ala Arg Pro Ser Leu Cys Thr Leu Val Pro Leu Gly Pro Glu Cys
1 5 10 15
Leu Arg Pro Phe Thr Arg Glu Ser Leu Ala Ala Ile Glu Gln Arg Ala
20 25 30
Val Glu Glu Glu Ala Arg Leu Gln Arg Asn Lys Gln Met Glu Ile Glu
35 40 45
Glu Pro Glu Arg Lys Pro Arg Ser Asp Leu Glu Ala Gly Lys Asn Leu
50 55 60
Pro Met Ile Tyr Gly Asp Pro Pro Pro Glu Val Ile Gly Ile Pro Leu
65 70 75 80
Glu Asp Leu Asp Pro Tyr Tyr Ser Asn Lys Lys Thr Phe Ile Val Leu
85 90 95
Asn Lys Gly Lys Ala Ile Phe Arg Phe Ser Ala Thr Pro Ala Leu Tyr
100 105 110
Leu Leu Ser Pro Phe Ser Val Val Arg Arg Gly Ala Ile Lys Val Leu
115 120 125
Ile His Ala Leu Phe Ser Met Phe Ile Met Ile Thr Ile Leu Thr Asn
130 135 140
Cys Val Phe Met Thr Met Ser Asp Pro Pro Pro Trp Ser Lys Asn Val
145 150 155 160
Glu Tyr Thr Phe Thr Gly Ile Tyr Thr Phe Glu Ser Leu Ile Lys Ile
165 170 175
Leu Ala Arg Gly Phe Cys Val Asp Asp Phe Thr Phe Leu Arg Asp Pro
180 185 190
Trp Asn Trp Leu Asp Phe Ser Val Ile Met Met Ala Tyr Leu Thr Glu
195 200 205
Phe Val Asp Leu Gly Asn Ile Ser Ala Leu Arg Thr Phe Arg Val Leu
210 215 220
Arg Ala Leu Lys Thr Ile Thr Val Ile Pro Gly Leu Lys Thr Ile Val
225 230 235 240
Gly Ala Leu Ile Gln Ser Val Lys Lys Leu Ser Asp Val Met Ile Leu
245 250 255
Thr Val Phe Cys Leu Ser Val Phe Ala Leu Val Gly Leu Gln Leu Phe
260 265 270
Met Gly Asn Leu Arg Gln Lys Cys Val Arg Trp Pro Pro Pro Phe Asn
275 280 285
Asp Thr Asn Thr Thr Trp Tyr Ser Asn Asp Thr Trp Tyr Gly Asn Asp
290 295 300
Thr Trp Tyr Gly Asn Glu Met Trp Tyr Gly Asn Asp Ser Trp Tyr Ala
305 310 315 320
Asn Asp Thr Trp Asn Ser His Ala Ser Trp Ala Thr Asn Asp Thr Phe
325 330 335
Asp Trp Asp Ala Tyr Ile Ser Asp Glu Gly Asn Phe Tyr Phe Leu Glu
340 345 350
Gly Ser Asn Asp Ala Leu Leu Cys Gly Asn Ser Ser Asp Ala Gly His
355 360 365
Cys Pro Glu Gly Tyr Glu Cys Ile Lys Thr Gly Arg Asn Pro Asn Tyr
370 375 380
Gly Tyr Thr Ser Tyr Asp Thr Phe Ser Trp Ala Phe Leu Ala Leu Phe
385 390 395 400
Arg Leu Met Thr Gln Asp Tyr Trp Glu Asn Leu Phe Gln Leu Thr Leu
405 410 415
Arg Ala Ala Gly Lys Thr Tyr Met Ile Phe Phe Val Val Ile Ile Phe
420 425 430
Leu Gly Ser Phe Tyr Leu Ile Asn Leu Ile Leu Ala Val Val Ala Met
435 440 445
Ala Tyr Ala Glu Gln Asn Glu Ala Thr Leu Ala Glu Asp Lys Glu Lys
450 455 460
Glu Glu Glu Phe Gln Gln Met Leu Glu Lys Phe Lys Lys His Gln Glu
465 470 475 480
Glu Leu Glu Lys Ala Lys Ala Ala Gln Ala Leu Glu Gly Gly Glu Ala
485 490 495
Asp Gly Asp Pro Ala His Gly Lys Asp Cys Asn Gly Ser Leu Asp Thr
500 505 510
Ser Gln Gly Glu Lys Gly Ala Pro Arg Gln Ser Ser Ser Gly Asp Ser
515 520 525
Gly Ile Ser Asp Ala Met Glu Glu Leu Glu Glu Ala His Gln Lys Cys
530 535 540
Pro Pro Trp Trp Tyr Lys Cys Ala His Lys Val Leu Ile Trp Asn Cys
545 550 555 560
Cys Ala Pro Trp Leu Lys Phe Lys Asn Ile Ile His Leu Ile Val Met
565 570 575
Asp Pro Phe Val Asp Leu Gly Ile Thr Ile Cys Ile Val Leu Asn Thr
580 585 590
Leu Phe Met Ala Met Glu His Tyr Pro Met Thr Glu His Phe Asp Asn
595 600 605
Val Leu Thr Val Gly Asn Leu Val Phe Thr Gly Ile Phe Thr Ala Glu
610 615 620
Met Val Leu Lys Leu Ile Ala Met Asp Pro Tyr Glu Tyr Phe Gln Gln
625 630 635 640
Gly Trp Asn Ile Phe Asp Ser Ile Ile Val Thr Leu Ser Leu Val Glu
645 650 655
Leu Gly Leu Ala Asn Val Gln Gly Leu Ser Val Leu Arg Ser Phe Arg
660 665 670
Leu Leu Arg Val Phe Lys Leu Ala Lys Ser Trp Pro Thr Leu Asn Met
675 680 685
Leu Ile Lys Ile Ile Gly Asn Ser Val Gly Ala Leu Gly Asn Leu Thr
690 695 700
Leu Val Leu Ala Ile Ile Val Phe Ile Phe Ala Val Val Gly Met Gln
705 710 715 720
Leu Phe Gly Lys Ser Tyr Lys Glu Cys Val Cys Lys Ile Ala Leu Asp
725 730 735
Cys Asn Leu Pro Arg Trp His Met His Asp Phe Phe His Ser Phe Leu
740 745 750
Ile Val Phe Arg Ile Leu Cys Gly Glu Trp Ile Glu Thr Met Trp Asp
755 760 765
Cys Met Glu Val Ala Gly Gln Ala Met Cys Leu Thr Val Phe Leu Met
770 775 780
Val Met Val Ile Gly Asn Leu Val Val Leu Asn Leu Phe Leu Ala Leu
785 790 795 800
Leu Leu Ser Ser Phe Ser Ala Asp Ser Leu Ala Ala Ser Asp Glu Asp
805 810 815
Gly Glu Met Asn Asn Leu Gln Ile Ala Ile Gly Arg Ile Lys Leu Gly
820 825 830
Ile Gly Phe Ala Lys Ala Phe Leu Leu Gly Leu Leu His Gly Lys Ile
835 840 845
Leu Ser Pro Lys Asp Ile Met Leu Ser Leu Gly Glu Ala Asp Gly Ala
850 855 860
Gly Glu Ala Gly Glu Ala Gly Glu Thr Ala Pro Glu Asp Glu Lys Lys
865 870 875 880
Glu Pro Pro Glu Glu Asp Leu Lys Lys Asp Asn His Ile Leu Asn His
885 890 895
Met Gly Leu Ala Asp Gly Pro Pro Ser Ser Leu Glu Leu Asp His Leu
900 905 910
Asn Phe Ile Asn Asn Pro Tyr Leu Thr Ile Gln Val Pro Ile Ala Ser
915 920 925
Glu Glu Ser Asp Leu Glu Met Pro Thr Glu Glu Glu Thr Asp Thr Phe
930 935 940
Ser Glu Pro Glu Asp Ser Lys Lys Pro Trp Pro Cys Leu Tyr Val Asp
945 950 955 960
Ile Ser Gln Gly Arg Gly Lys Lys Trp Trp Thr Leu Arg Arg Ala Cys
965 970 975
Phe Lys Ile Val Glu His Asn Trp Phe Glu Thr Phe Ile Val Phe Met
980 985 990
Ile Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu
995 1000 1005
Gln Arg Arg Val Ile Arg Thr Ile Leu Glu Tyr Ala Asp Lys Val
1010 1015 1020
Phe Thr Tyr Ile Phe Ile Met Glu Met Leu Leu Lys Trp Val Ala
1025 1030 1035
Tyr Gly Phe Lys Val Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp
1040 1045 1050
Phe Leu Ile Val Asp Val Ser Ile Ile Ser Leu Val Ala Asn Trp
1055 1060 1065
Leu Gly Tyr Ser Glu Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu
1070 1075 1080
Arg Ala Leu Arg Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met
1085 1090 1095
Arg Val Val Val Asn Ala Leu Leu Gly Ala Ile Pro Ser Ile Met
1100 1105 1110
Asn Val Leu Leu Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile
1115 1120 1125
Met Gly Val Asn Leu Phe Ala Gly Lys Phe Tyr Tyr Cys Ile Asn
1130 1135 1140
Thr Thr Thr Ser Glu Arg Phe Asp Ile Ser Glu Val Asn Asn Lys
1145 1150 1155
Ser Glu Cys Glu Ser Leu Met His Thr Gly Gln Val Arg Trp Leu
1160 1165 1170
Asn Val Lys Val Asn Tyr Asp Asn Val Gly Leu Gly Tyr Leu Ser
1175 1180 1185
Leu Leu Gln Val Ala Thr Phe Lys Gly Trp Met Asp Ile Met Tyr
1190 1195 1200
Ala Ala Val Asp Ser Arg Glu Lys Glu Glu Gln Pro Gln Tyr Glu
1205 1210 1215
Val Asn Leu Tyr Met Tyr Leu Tyr Phe Val Ile Phe Ile Ile Phe
1220 1225 1230
Gly Ser Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp
1235 1240 1245
Asn Phe Asn Gln Gln Lys Lys Lys Leu Gly Gly Lys Asp Ile Phe
1250 1255 1260
Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu
1265 1270 1275
Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg Pro Gln Asn Lys
1280 1285 1290
Ile Gln Gly Met Val Tyr Asp Leu Val Thr Lys Gln Ala Phe Asp
1295 1300 1305
Ile Thr Ile Met Ile Leu Ile Cys Leu Asn Met Val Thr Met Met
1310 1315 1320
Val Glu Thr Asp Asn Gln Ser Gln Leu Lys Val Asp Ile Leu Tyr
1325 1330 1335
Asn Ile Asn Met Ile Phe Ile Ile Ile Phe Thr Gly Glu Cys Val
1340 1345 1350
Leu Lys Met Leu Ala Leu Arg Gln Tyr Tyr Phe Thr Val Gly Trp
1355 1360 1365
Asn Ile Phe Asp Phe Val Val Val Ile Leu Ser Ile Val Gly Leu
1370 1375 1380
Ala Leu Ser Asp Leu Ile Gln Lys Tyr Phe Val Ser Pro Thr Leu
1385 1390 1395
Phe Arg Val Ile Arg Leu Ala Arg Ile Gly Arg Val Leu Arg Leu
1400 1405 1410
Ile Arg Gly Ala Lys Gly Ile Arg Thr Leu Leu Phe Ala Leu Met
1415 1420 1425
Met Ser Leu Pro Ala Leu Phe Asn Ile Gly Leu Leu Leu Phe Leu
1430 1435 1440
Val Met Phe Ile Tyr Ser Ile Phe Gly Met Ser Asn Phe Ala Tyr
1445 1450 1455
Val Lys Lys Glu Ser Gly Ile Asp Asp Met Phe Asn Phe Glu Thr
1460 1465 1470
Phe Gly Asn Ser Ile Ile Cys Leu Phe Glu Ile Thr Thr Ser Ala
1475 1480 1485
Gly Trp Asp Gly Leu Leu Asn Pro Ile Leu Asn Ser Gly Pro Pro
1490 1495 1500
Asp Cys Asp Pro Asn Leu Glu Asn Pro Gly Thr Ser Val Lys Gly
1505 1510 1515
Asp Cys Gly Asn Pro Ser Ile Gly Ile Cys Phe Phe Cys Ser Tyr
1520 1525 1530
Ile Ile Ile Ser Phe Leu Ile Val Val Asn Met Tyr Ile Ala Ile
1535 1540 1545
Ile Leu Glu Asn Phe Asn Val Ala Thr Glu Glu Ser Ser Glu Pro
1550 1555 1560
Leu Gly Glu Asp Asp Phe Glu Met Phe Tyr Glu Thr Trp Glu Lys
1565 1570 1575
Phe Asp Pro Asp Ala Thr Gln Phe Ile Ala Tyr Ser Arg Leu Ser
1580 1585 1590
Asp Phe Val Asp Thr Leu Gln Glu Pro Leu Arg Ile Ala Lys Pro
1595 1600 1605
Asn Lys Ile Lys Leu Ile Thr Leu Asp Leu Pro Met Val Pro Gly
1610 1615 1620
Asp Lys Ile His Cys Leu Asp Ile Leu Phe Ala Leu Thr Lys Glu
1625 1630 1635
Val Leu Gly Asp Ser Gly Glu Met Asp Ala Leu Lys Gln Thr Met
1640 1645 1650
Glu Glu Lys Phe Met Ala Ala Asn Pro Ser Lys Val Ser Tyr Glu
1655 1660 1665
Pro Ile Thr Thr Thr Leu Lys Arg Lys His Glu Glu Val Cys Ala
1670 1675 1680
Ile Lys Ile Gln Arg Ala Tyr Arg Arg His Leu Leu Gln Arg Ser
1685 1690 1695
Met Lys Gln Ala Ser Tyr Met Tyr Arg His Ser His Asp Gly Ser
1700 1705 1710
Gly Asp Asp Ala Pro Glu Lys Glu Gly Leu Leu Ala Asn Thr Met
1715 1720 1725
Ser Lys Met Tyr Gly His Glu Asn Gly Asn Ser Ser Ser Pro Ser
1730 1735 1740
Pro Glu Glu Lys Gly Glu Ala Gly Asp Ala Gly Pro Thr Met Gly
1745 1750 1755
Leu Met Pro Ile Ser Pro Ser Asp Thr Ala Trp Pro Pro Ala Pro
1760 1765 1770
Pro Pro Gly Gln Thr Val Arg Pro Gly Val Lys Glu Ser Leu Val
1775 1780 1785
<210> 32
<211> 2016
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(2016)
<223> 人 Nav 1.5（图 9）
<400> 32
Met Ala Asn Phe Leu Leu Pro Arg Gly Thr Ser Ser Phe Arg Arg Phe
1 5 10 15
Thr Arg Glu Ser Leu Ala Ala Ile Glu Lys Arg Met Ala Glu Lys Gln
20 25 30
Ala Arg Gly Ser Thr Thr Leu Gln Glu Ser Arg Glu Gly Leu Pro Glu
35 40 45
Glu Glu Ala Pro Arg Pro Gln Leu Asp Leu Gln Ala Ser Lys Lys Leu
50 55 60
Pro Asp Leu Tyr Gly Asn Pro Pro Gln Glu Leu Ile Gly Glu Pro Leu
65 70 75 80
Glu Asp Leu Asp Pro Phe Tyr Ser Thr Gln Lys Thr Phe Ile Val Leu
85 90 95
Asn Lys Gly Lys Thr Ile Phe Arg Phe Ser Ala Thr Asn Ala Leu Tyr
100 105 110
Val Leu Ser Pro Phe His Pro Ile Arg Arg Ala Ala Val Lys Ile Leu
115 120 125
Val His Ser Leu Phe Asn Met Leu Ile Met Cys Thr Ile Leu Thr Asn
130 135 140
Cys Val Phe Met Ala Gln His Asp Pro Pro Pro Trp Thr Lys Tyr Val
145 150 155 160
Glu Tyr Thr Phe Thr Ala Ile Tyr Thr Phe Glu Ser Leu Val Lys Ile
165 170 175
Leu Ala Arg Gly Phe Cys Leu His Ala Phe Thr Phe Leu Arg Asp Pro
180 185 190
Trp Asn Trp Leu Asp Phe Ser Val Ile Ile Met Ala Tyr Thr Thr Glu
195 200 205
Phe Val Asp Leu Gly Asn Val Ser Ala Leu Arg Thr Phe Arg Val Leu
210 215 220
Arg Ala Leu Lys Thr Ile Ser Val Ile Ser Gly Leu Lys Thr Ile Val
225 230 235 240
Gly Ala Leu Ile Gln Ser Val Lys Lys Leu Ala Asp Val Met Val Leu
245 250 255
Thr Val Phe Cys Leu Ser Val Phe Ala Leu Ile Gly Leu Gln Leu Phe
260 265 270
Met Gly Asn Leu Arg His Lys Cys Val Arg Asn Phe Thr Ala Leu Asn
275 280 285
Gly Thr Asn Gly Ser Val Glu Ala Asp Gly Leu Val Trp Glu Ser Leu
290 295 300
Asp Leu Tyr Leu Ser Asp Pro Glu Asn Tyr Leu Leu Lys Asn Gly Thr
305 310 315 320
Ser Asp Val Leu Leu Cys Gly Asn Ser Ser Asp Ala Gly Thr Cys Pro
325 330 335
Glu Gly Tyr Arg Cys Leu Lys Ala Gly Glu Asn Pro Asp His Gly Tyr
340 345 350
Thr Ser Phe Asp Ser Phe Ala Trp Ala Phe Leu Ala Leu Phe Arg Leu
355 360 365
Met Thr Gln Asp Cys Trp Glu Arg Leu Tyr Gln Gln Thr Leu Arg Ser
370 375 380
Ala Gly Lys Ile Tyr Met Ile Phe Phe Met Leu Val Ile Phe Leu Gly
385 390 395 400
Ser Phe Tyr Leu Val Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr
405 410 415
Glu Glu Gln Asn Gln Ala Thr Ile Ala Glu Thr Glu Glu Lys Glu Lys
420 425 430
Arg Phe Gln Glu Ala Met Glu Met Leu Lys Lys Glu His Glu Ala Leu
435 440 445
Thr Ile Arg Gly Val Asp Thr Val Ser Arg Ser Ser Leu Glu Met Ser
450 455 460
Pro Leu Ala Pro Val Asn Ser His Glu Arg Arg Ser Lys Arg Arg Lys
465 470 475 480
Arg Met Ser Ser Gly Thr Glu Glu Cys Gly Glu Asp Arg Leu Pro Lys
485 490 495
Ser Asp Ser Glu Asp Gly Pro Arg Ala Met Asn His Leu Ser Leu Thr
500 505 510
Arg Gly Leu Ser Arg Thr Ser Met Lys Pro Arg Ser Ser Arg Gly Ser
515 520 525
Ile Phe Thr Phe Arg Arg Arg Asp Leu Gly Ser Glu Ala Asp Phe Ala
530 535 540
Asp Asp Glu Asn Ser Thr Ala Gly Glu Ser Glu Ser His His Thr Ser
545 550 555 560
Leu Leu Val Pro Trp Pro Leu Arg Arg Thr Ser Ala Gln Gly Gln Pro
565 570 575
Ser Pro Gly Thr Ser Ala Pro Gly His Ala Leu His Gly Lys Lys Asn
580 585 590
Ser Thr Val Asp Cys Asn Gly Val Val Ser Leu Leu Gly Ala Gly Asp
595 600 605
Pro Glu Ala Thr Ser Pro Gly Ser His Leu Leu Arg Pro Val Met Leu
610 615 620
Glu His Pro Pro Asp Thr Thr Thr Pro Ser Glu Glu Pro Gly Gly Pro
625 630 635 640
Gln Met Leu Thr Ser Gln Ala Pro Cys Val Asp Gly Phe Glu Glu Pro
645 650 655
Gly Ala Arg Gln Arg Ala Leu Ser Ala Val Ser Val Leu Thr Ser Ala
660 665 670
Leu Glu Glu Leu Glu Glu Ser Arg His Lys Cys Pro Pro Cys Trp Asn
675 680 685
Arg Leu Ala Gln Arg Tyr Leu Ile Trp Glu Cys Cys Pro Leu Trp Met
690 695 700
Ser Ile Lys Gln Gly Val Lys Leu Val Val Met Asp Pro Phe Thr Asp
705 710 715 720
Leu Thr Ile Thr Met Cys Ile Val Leu Asn Thr Leu Phe Met Ala Leu
725 730 735
Glu His Tyr Asn Met Thr Ser Glu Phe Glu Glu Met Leu Gln Val Gly
740 745 750
Asn Leu Val Phe Thr Gly Ile Phe Thr Ala Glu Met Thr Phe Lys Ile
755 760 765
Ile Ala Leu Asp Pro Tyr Tyr Tyr Phe Gln Gln Gly Trp Asn Ile Phe
770 775 780
Asp Ser Ile Ile Val Ile Leu Ser Leu Met Glu Leu Gly Leu Ser Arg
785 790 795 800
Met Ser Asn Leu Ser Val Leu Arg Ser Phe Arg Leu Leu Arg Val Phe
805 810 815
Lys Leu Ala Lys Ser Trp Pro Thr Leu Asn Thr Leu Ile Lys Ile Ile
820 825 830
Gly Asn Ser Val Gly Ala Leu Gly Asn Leu Thr Leu Val Leu Ala Ile
835 840 845
Ile Val Phe Ile Phe Ala Val Val Gly Met Gln Leu Phe Gly Lys Asn
850 855 860
Tyr Ser Glu Leu Arg Asp Ser Asp Ser Gly Leu Leu Pro Arg Trp His
865 870 875 880
Met Met Asp Phe Phe His Ala Phe Leu Ile Ile Phe Arg Ile Leu Cys
885 890 895
Gly Glu Trp Ile Glu Thr Met Trp Asp Cys Met Glu Val Ser Gly Gln
900 905 910
Ser Leu Cys Leu Leu Val Phe Leu Leu Val Met Val Ile Gly Asn Leu
915 920 925
Val Val Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe Ser Ala
930 935 940
Asp Asn Leu Thr Ala Pro Asp Glu Asp Arg Glu Met Asn Asn Leu Gln
945 950 955 960
Leu Ala Leu Ala Arg Ile Gln Arg Gly Leu Arg Phe Val Lys Arg Thr
965 970 975
Thr Trp Asp Phe Cys Cys Gly Leu Leu Arg Gln Arg Pro Gln Lys Pro
980 985 990
Ala Ala Leu Ala Ala Gln Gly Gln Leu Pro Ser Cys Ile Ala Thr Pro
995 1000 1005
Tyr Ser Pro Pro Pro Pro Glu Thr Glu Lys Val Pro Pro Thr Arg
1010 1015 1020
Lys Glu Thr Arg Phe Glu Glu Gly Glu Gln Pro Gly Gln Gly Thr
1025 1030 1035
Pro Gly Asp Pro Glu Pro Val Cys Val Pro Ile Ala Val Ala Glu
1040 1045 1050
Ser Asp Thr Asp Asp Gln Glu Glu Asp Glu Glu Asn Ser Leu Gly
1055 1060 1065
Thr Glu Glu Glu Ser Ser Lys Gln Gln Glu Ser Gln Pro Val Ser
1070 1075 1080
Gly Gly Pro Glu Ala Pro Pro Asp Ser Arg Thr Trp Ser Gln Val
1085 1090 1095
Ser Ala Thr Ala Ser Ser Glu Ala Glu Ala Ser Ala Ser Gln Ala
1100 1105 1110
Asp Trp Arg Gln Gln Trp Lys Ala Glu Pro Gln Ala Pro Gly Cys
1115 1120 1125
Gly Glu Thr Pro Glu Asp Ser Cys Ser Glu Gly Ser Thr Ala Asp
1130 1135 1140
Met Thr Asn Thr Ala Glu Leu Leu Glu Gln Ile Pro Asp Leu Gly
1145 1150 1155
Gln Asp Val Lys Asp Pro Glu Asp Cys Phe Thr Glu Gly Cys Val
1160 1165 1170
Arg Arg Cys Pro Cys Cys Ala Val Asp Thr Thr Gln Ala Pro Gly
1175 1180 1185
Lys Val Trp Trp Arg Leu Arg Lys Thr Cys Tyr His Ile Val Glu
1190 1195 1200
His Ser Trp Phe Glu Thr Phe Ile Ile Phe Met Ile Leu Leu Ser
1205 1210 1215
Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Leu Glu Glu Arg Lys
1220 1225 1230
Thr Ile Lys Val Leu Leu Glu Tyr Ala Asp Lys Met Phe Thr Tyr
1235 1240 1245
Val Phe Val Leu Glu Met Leu Leu Lys Trp Val Ala Tyr Gly Phe
1250 1255 1260
Lys Lys Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile
1265 1270 1275
Val Asp Val Ser Leu Val Ser Leu Val Ala Asn Thr Leu Gly Phe
1280 1285 1290
Ala Glu Met Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu
1295 1300 1305
Arg Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val
1310 1315 1320
Val Asn Ala Leu Val Gly Ala Ile Pro Ser Ile Met Asn Val Leu
1325 1330 1335
Leu Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile Met Gly Val
1340 1345 1350
Asn Leu Phe Ala Gly Lys Phe Gly Arg Cys Ile Asn Gln Thr Glu
1355 1360 1365
Gly Asp Leu Pro Leu Asn Tyr Thr Ile Val Asn Asn Lys Ser Gln
1370 1375 1380
Cys Glu Ser Leu Asn Leu Thr Gly Glu Leu Tyr Trp Thr Lys Val
1385 1390 1395
Lys Val Asn Phe Asp Asn Val Gly Ala Gly Tyr Leu Ala Leu Leu
1400 1405 1410
Gln Val Ala Thr Phe Lys Gly Trp Met Asp Ile Met Tyr Ala Ala
1415 1420 1425
Val Asp Ser Arg Gly Tyr Glu Glu Gln Pro Gln Trp Glu Tyr Asn
1430 1435 1440
Leu Tyr Met Tyr Ile Tyr Phe Val Ile Phe Ile Ile Phe Gly Ser
1445 1450 1455
Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe
1460 1465 1470
Asn Gln Gln Lys Lys Lys Leu Gly Gly Gln Asp Ile Phe Met Thr
1475 1480 1485
Glu Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser
1490 1495 1500
Lys Lys Pro Gln Lys Pro Ile Pro Arg Pro Leu Asn Lys Tyr Gln
1505 1510 1515
Gly Phe Ile Phe Asp Ile Val Thr Lys Gln Ala Phe Asp Val Thr
1520 1525 1530
Ile Met Phe Leu Ile Cys Leu Asn Met Val Thr Met Met Val Glu
1535 1540 1545
Thr Asp Asp Gln Ser Pro Glu Lys Ile Asn Ile Leu Ala Lys Ile
1550 1555 1560
Asn Leu Leu Phe Val Ala Ile Phe Thr Gly Glu Cys Ile Val Lys
1565 1570 1575
Leu Ala Ala Leu Arg His Tyr Tyr Phe Thr Asn Ser Trp Asn Ile
1580 1585 1590
Phe Asp Phe Val Val Val Ile Leu Ser Ile Val Gly Thr Val Leu
1595 1600 1605
Ser Asp Ile Ile Gln Lys Tyr Phe Phe Ser Pro Thr Leu Phe Arg
1610 1615 1620
Val Ile Arg Leu Ala Arg Ile Gly Arg Ile Leu Arg Leu Ile Arg
1625 1630 1635
Gly Ala Lys Gly Ile Arg Thr Leu Leu Phe Ala Leu Met Met Ser
1640 1645 1650
Leu Pro Ala Leu Phe Asn Ile Gly Leu Leu Leu Phe Leu Val Met
1655 1660 1665
Phe Ile Tyr Ser Ile Phe Gly Met Ala Asn Phe Ala Tyr Val Lys
1670 1675 1680
Trp Glu Ala Gly Ile Asp Asp Met Phe Asn Phe Gln Thr Phe Ala
1685 1690 1695
Asn Ser Met Leu Cys Leu Phe Gln Ile Thr Thr Ser Ala Gly Trp
1700 1705 1710
Asp Gly Leu Leu Ser Pro Ile Leu Asn Thr Gly Pro Pro Tyr Cys
1715 1720 1725
Asp Pro Thr Leu Pro Asn Ser Asn Gly Ser Arg Gly Asp Cys Gly
1730 1735 1740
Ser Pro Ala Val Gly Ile Leu Phe Phe Thr Thr Tyr Ile Ile Ile
1745 1750 1755
Ser Phe Leu Ile Val Val Asn Met Tyr Ile Ala Ile Ile Leu Glu
1760 1765 1770
Asn Phe Ser Val Ala Thr Glu Glu Ser Thr Glu Pro Leu Ser Glu
1775 1780 1785
Asp Asp Phe Asp Met Phe Tyr Glu Ile Trp Glu Lys Phe Asp Pro
1790 1795 1800
Glu Ala Thr Gln Phe Ile Glu Tyr Ser Val Leu Ser Asp Phe Ala
1805 1810 1815
Asp Ala Leu Ser Glu Pro Leu Arg Ile Ala Lys Pro Asn Gln Ile
1820 1825 1830
Ser Leu Ile Asn Met Asp Leu Pro Met Val Ser Gly Asp Arg Ile
1835 1840 1845
His Cys Met Asp Ile Leu Phe Ala Phe Thr Lys Arg Val Leu Gly
1850 1855 1860
Glu Ser Gly Glu Met Asp Ala Leu Lys Ile Gln Met Glu Glu Lys
1865 1870 1875
Phe Met Ala Ala Asn Pro Ser Lys Ile Ser Tyr Glu Pro Ile Thr
1880 1885 1890
Thr Thr Leu Arg Arg Lys His Glu Glu Val Ser Ala Met Val Ile
1895 1900 1905
Gln Arg Ala Phe Arg Arg His Leu Leu Gln Arg Ser Leu Lys His
1910 1915 1920
Ala Ser Phe Leu Phe Arg Gln Gln Ala Gly Ser Gly Leu Ser Glu
1925 1930 1935
Glu Asp Ala Pro Glu Arg Glu Gly Leu Ile Ala Tyr Val Met Ser
1940 1945 1950
Glu Asn Phe Ser Arg Pro Leu Gly Pro Pro Ser Ser Ser Ser Ile
1955 1960 1965
Ser Ser Thr Ser Phe Pro Pro Ser Tyr Asp Ser Val Thr Arg Ala
1970 1975 1980
Thr Ser Asp Asn Leu Gln Val Arg Gly Ser Asp Tyr Ser His Ser
1985 1990 1995
Glu Asp Leu Ala Asp Phe Pro Pro Ser Pro Asp Arg Asp Arg Glu
2000 2005 2010
Ser Ile Val
2015
<210> 33
<211> 1980
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(1980)
<223> 人 Nav 1.6（图 9）
<400> 33
Met Ala Ala Arg Leu Leu Ala Pro Pro Gly Pro Asp Ser Phe Lys Pro
1 5 10 15
Phe Thr Pro Glu Ser Leu Ala Asn Ile Glu Arg Arg Ile Ala Glu Ser
20 25 30
Lys Leu Lys Lys Pro Pro Lys Ala Asp Gly Ser His Arg Glu Asp Asp
35 40 45
Glu Asp Ser Lys Pro Lys Pro Asn Ser Asp Leu Glu Ala Gly Lys Ser
50 55 60
Leu Pro Phe Ile Tyr Gly Asp Ile Pro Gln Gly Leu Val Ala Val Pro
65 70 75 80
Leu Glu Asp Phe Asp Pro Tyr Tyr Leu Thr Gln Lys Thr Phe Val Val
85 90 95
Leu Asn Arg Gly Lys Thr Leu Phe Arg Phe Ser Ala Thr Pro Ala Leu
100 105 110
Tyr Ile Leu Ser Pro Phe Asn Leu Ile Arg Arg Ile Ala Ile Lys Ile
115 120 125
Leu Ile His Ser Val Phe Ser Met Ile Ile Met Cys Thr Ile Leu Thr
130 135 140
Asn Cys Val Phe Met Thr Phe Ser Asn Pro Pro Asp Trp Ser Lys Asn
145 150 155 160
Val Glu Tyr Thr Phe Thr Gly Ile Tyr Thr Phe Glu Ser Leu Val Lys
165 170 175
Ile Ile Ala Arg Gly Phe Cys Ile Asp Gly Phe Thr Phe Leu Arg Asp
180 185 190
Pro Trp Asn Trp Leu Asp Phe Ser Val Ile Met Met Ala Tyr Ile Thr
195 200 205
Glu Phe Val Asn Leu Gly Asn Val Ser Ala Leu Arg Thr Phe Arg Val
210 215 220
Leu Arg Ala Leu Lys Thr Ile Ser Val Ile Pro Gly Leu Lys Thr Ile
225 230 235 240
Val Gly Ala Leu Ile Gln Ser Val Lys Lys Leu Ser Asp Val Met Ile
245 250 255
Leu Thr Val Phe Cys Leu Ser Val Phe Ala Leu Ile Gly Leu Gln Leu
260 265 270
Phe Met Gly Asn Leu Arg Asn Lys Cys Val Val Trp Pro Ile Asn Phe
275 280 285
Asn Glu Ser Tyr Leu Glu Asn Gly Thr Lys Gly Phe Asp Trp Glu Glu
290 295 300
Tyr Ile Asn Asn Lys Thr Asn Phe Tyr Thr Val Pro Gly Met Leu Glu
305 310 315 320
Pro Leu Leu Cys Gly Asn Ser Ser Asp Ala Gly Gln Cys Pro Glu Gly
325 330 335
Tyr Gln Cys Met Lys Ala Gly Arg Asn Pro Asn Tyr Gly Tyr Thr Ser
340 345 350
Phe Asp Thr Phe Ser Trp Ala Phe Leu Ala Leu Phe Arg Leu Met Thr
355 360 365
Gln Asp Tyr Trp Glu Asn Leu Tyr Gln Leu Thr Leu Arg Ala Ala Gly
370 375 380
Lys Thr Tyr Met Ile Phe Phe Val Leu Val Ile Phe Val Gly Ser Phe
385 390 395 400
Tyr Leu Val Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr Glu Glu
405 410 415
Gln Asn Gln Ala Thr Leu Glu Glu Ala Glu Gln Lys Glu Ala Glu Phe
420 425 430
Lys Ala Met Leu Glu Gln Leu Lys Lys Gln Gln Glu Glu Ala Gln Ala
435 440 445
Ala Ala Met Ala Thr Ser Ala Gly Thr Val Ser Glu Asp Ala Ile Glu
450 455 460
Glu Glu Gly Glu Glu Gly Gly Gly Ser Pro Arg Ser Ser Ser Glu Ile
465 470 475 480
Ser Lys Leu Ser Ser Lys Ser Ala Lys Glu Arg Arg Asn Arg Arg Lys
485 490 495
Lys Arg Lys Gln Lys Glu Leu Ser Glu Gly Glu Glu Lys Gly Asp Pro
500 505 510
Glu Lys Val Phe Lys Ser Glu Ser Glu Asp Gly Met Arg Arg Lys Ala
515 520 525
Phe Arg Leu Pro Asp Asn Arg Ile Gly Arg Lys Phe Ser Ile Met Asn
530 535 540
Gln Ser Leu Leu Ser Ile Pro Gly Ser Pro Phe Leu Ser Arg His Asn
545 550 555 560
Ser Lys Ser Ser Ile Phe Ser Phe Arg Gly Pro Gly Arg Phe Arg Asp
565 570 575
Pro Gly Ser Glu Asn Glu Phe Ala Asp Asp Glu His Ser Thr Val Glu
580 585 590
Glu Ser Glu Gly Arg Arg Asp Ser Leu Phe Ile Pro Ile Arg Ala Arg
595 600 605
Glu Arg Arg Ser Ser Tyr Ser Gly Tyr Ser Gly Tyr Ser Gln Gly Ser
610 615 620
Arg Ser Ser Arg Ile Phe Pro Ser Leu Arg Arg Ser Val Lys Arg Asn
625 630 635 640
Ser Thr Val Asp Cys Asn Gly Val Val Ser Leu Ile Gly Gly Pro Gly
645 650 655
Ser His Ile Gly Gly Arg Leu Leu Pro Glu Ala Thr Thr Glu Val Glu
660 665 670
Ile Lys Lys Lys Gly Pro Gly Ser Leu Leu Val Ser Met Asp Gln Leu
675 680 685
Ala Ser Tyr Gly Arg Lys Asp Arg Ile Asn Ser Ile Met Ser Val Val
690 695 700
Thr Asn Thr Leu Val Glu Glu Leu Glu Glu Ser Gln Arg Lys Cys Pro
705 710 715 720
Pro Cys Trp Tyr Lys Phe Ala Asn Thr Phe Leu Ile Trp Glu Cys His
725 730 735
Pro Tyr Trp Ile Lys Leu Lys Glu Ile Val Asn Leu Ile Val Met Asp
740 745 750
Pro Phe Val Asp Leu Ala Ile Thr Ile Cys Ile Val Leu Asn Thr Leu
755 760 765
Phe Met Ala Met Glu His His Pro Met Thr Pro Gln Phe Glu His Val
770 775 780
Leu Ala Val Gly Asn Leu Val Phe Thr Gly Ile Phe Thr Ala Glu Met
785 790 795 800
Phe Leu Lys Leu Ile Ala Met Asp Pro Tyr Tyr Tyr Phe Gln Glu Gly
805 810 815
Trp Asn Ile Phe Asp Gly Phe Ile Val Ser Leu Ser Leu Met Glu Leu
820 825 830
Ser Leu Ala Asp Val Glu Gly Leu Ser Val Leu Arg Ser Phe Arg Leu
835 840 845
Leu Arg Val Phe Lys Leu Ala Lys Ser Trp Pro Thr Leu Asn Met Leu
850 855 860
Ile Lys Ile Ile Gly Asn Ser Val Gly Ala Leu Gly Asn Leu Thr Leu
865 870 875 880
Val Leu Ala Ile Ile Val Phe Ile Phe Ala Val Val Gly Met Gln Leu
885 890 895
Phe Gly Lys Ser Tyr Lys Glu Cys Val Cys Lys Ile Asn Gln Asp Cys
900 905 910
Glu Leu Pro Arg Trp His Met His Asp Phe Phe His Ser Phe Leu Ile
915 920 925
Val Phe Arg Val Leu Cys Gly Glu Trp Ile Glu Thr Met Trp Asp Cys
930 935 940
Met Glu Val Ala Gly Gln Ala Met Cys Leu Ile Val Phe Met Met Val
945 950 955 960
Met Val Ile Gly Asn Leu Val Val Leu Asn Leu Phe Leu Ala Leu Leu
965 970 975
Leu Ser Ser Phe Ser Ala Asp Asn Leu Ala Ala Thr Asp Asp Asp Gly
980 985 990
Glu Met Asn Asn Leu Gln Ile Ser Val Ile Arg Ile Lys Lys Gly Val
995 1000 1005
Ala Trp Thr Lys Leu Lys Val His Ala Phe Met Gln Ala His Phe
1010 1015 1020
Lys Gln Arg Glu Ala Asp Glu Val Lys Pro Leu Asp Glu Leu Tyr
1025 1030 1035
Glu Lys Lys Ala Asn Cys Ile Ala Asn His Thr Gly Ala Asp Ile
1040 1045 1050
His Arg Asn Gly Asp Phe Gln Lys Asn Gly Asn Gly Thr Thr Ser
1055 1060 1065
Gly Ile Gly Ser Ser Val Glu Lys Tyr Ile Ile Asp Glu Asp His
1070 1075 1080
Met Ser Phe Ile Asn Asn Pro Asn Leu Thr Val Arg Val Pro Ile
1085 1090 1095
Ala Val Gly Glu Ser Asp Phe Glu Asn Leu Asn Thr Glu Asp Val
1100 1105 1110
Ser Ser Glu Ser Asp Pro Glu Gly Ser Lys Asp Lys Leu Asp Asp
1115 1120 1125
Thr Ser Ser Ser Glu Gly Ser Thr Ile Asp Ile Lys Pro Glu Val
1130 1135 1140
Glu Glu Val Pro Val Glu Gln Pro Glu Glu Tyr Leu Asp Pro Asp
1145 1150 1155
Ala Cys Phe Thr Glu Gly Cys Val Gln Arg Phe Lys Cys Cys Gln
1160 1165 1170
Val Asn Ile Glu Glu Gly Leu Gly Lys Ser Trp Trp Ile Leu Arg
1175 1180 1185
Lys Thr Cys Phe Leu Ile Val Glu His Asn Trp Phe Glu Thr Phe
1190 1195 1200
Ile Ile Phe Met Ile Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu
1205 1210 1215
Asp Ile Tyr Ile Glu Gln Arg Lys Thr Ile Arg Thr Ile Leu Glu
1220 1225 1230
Tyr Ala Asp Lys Val Phe Thr Tyr Ile Phe Ile Leu Glu Met Leu
1235 1240 1245
Leu Lys Trp Thr Ala Tyr Gly Phe Val Lys Phe Phe Thr Asn Ala
1250 1255 1260
Trp Cys Trp Leu Asp Phe Leu Ile Val Ala Val Ser Leu Val Ser
1265 1270 1275
Leu Ile Ala Asn Ala Leu Gly Tyr Ser Glu Leu Gly Ala Ile Lys
1280 1285 1290
Ser Leu Arg Thr Leu Arg Ala Leu Arg Pro Leu Arg Ala Leu Ser
1295 1300 1305
Arg Phe Glu Gly Met Arg Val Val Val Asn Ala Leu Val Gly Ala
1310 1315 1320
Ile Pro Ser Ile Met Asn Val Leu Leu Val Cys Leu Ile Phe Trp
1325 1330 1335
Leu Ile Phe Ser Ile Met Gly Val Asn Leu Phe Ala Gly Lys Tyr
1340 1345 1350
His Tyr Cys Phe Asn Glu Thr Ser Glu Ile Arg Phe Glu Ile Glu
1355 1360 1365
Asp Val Asn Asn Lys Thr Glu Cys Glu Lys Leu Met Glu Gly Asn
1370 1375 1380
Asn Thr Glu Ile Arg Trp Lys Asn Val Lys Ile Asn Phe Asp Asn
1385 1390 1395
Val Gly Ala Gly Tyr Leu Ala Leu Leu Gln Val Ala Thr Phe Lys
1400 1405 1410
Gly Trp Met Asp Ile Met Tyr Ala Ala Val Asp Ser Arg Lys Pro
1415 1420 1425
Asp Glu Gln Pro Lys Tyr Glu Asp Asn Ile Tyr Met Tyr Ile Tyr
1430 1435 1440
Phe Val Ile Phe Ile Ile Phe Gly Ser Phe Phe Thr Leu Asn Leu
1445 1450 1455
Phe Ile Gly Val Ile Ile Asp Asn Phe Asn Gln Gln Lys Lys Lys
1460 1465 1470
Phe Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr
1475 1480 1485
Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro
1490 1495 1500
Ile Pro Arg Pro Leu Asn Lys Ile Gln Gly Ile Val Phe Asp Phe
1505 1510 1515
Val Thr Gln Gln Ala Phe Asp Ile Val Ile Met Met Leu Ile Cys
1520 1525 1530
Leu Asn Met Val Thr Met Met Val Glu Thr Asp Thr Gln Ser Lys
1535 1540 1545
Gln Met Glu Asn Ile Leu Tyr Trp Ile Asn Leu Val Phe Val Ile
1550 1555 1560
Phe Phe Thr Cys Glu Cys Val Leu Lys Met Phe Ala Leu Arg His
1565 1570 1575
Tyr Tyr Phe Thr Ile Gly Trp Asn Ile Phe Asp Phe Val Val Val
1580 1585 1590
Ile Leu Ser Ile Val Gly Met Phe Leu Ala Asp Ile Ile Glu Lys
1595 1600 1605
Tyr Phe Val Ser Pro Thr Leu Phe Arg Val Ile Arg Leu Ala Arg
1610 1615 1620
Ile Gly Arg Ile Leu Arg Leu Ile Lys Gly Ala Lys Gly Ile Arg
1625 1630 1635
Thr Leu Leu Phe Ala Leu Met Met Ser Leu Pro Ala Leu Phe Asn
1640 1645 1650
Ile Gly Leu Leu Leu Phe Leu Val Met Phe Ile Phe Ser Ile Phe
1655 1660 1665
Gly Met Ser Asn Phe Ala Tyr Val Lys His Glu Ala Gly Ile Asp
1670 1675 1680
Asp Met Phe Asn Phe Glu Thr Phe Gly Asn Ser Met Ile Cys Leu
1685 1690 1695
Phe Gln Ile Thr Thr Ser Ala Gly Trp Asp Gly Leu Leu Leu Pro
1700 1705 1710
Ile Leu Asn Arg Pro Pro Asp Cys Ser Leu Asp Lys Glu His Pro
1715 1720 1725
Gly Ser Gly Phe Lys Gly Asp Cys Gly Asn Pro Ser Val Gly Ile
1730 1735 1740
Phe Phe Phe Val Ser Tyr Ile Ile Ile Ser Phe Leu Ile Val Val
1745 1750 1755
Asn Met Tyr Ile Ala Ile Ile Leu Glu Asn Phe Ser Val Ala Thr
1760 1765 1770
Glu Glu Ser Ala Asp Pro Leu Ser Glu Asp Asp Phe Glu Thr Phe
1775 1780 1785
Tyr Glu Ile Trp Glu Lys Phe Asp Pro Asp Ala Thr Gln Phe Ile
1790 1795 1800
Glu Tyr Cys Lys Leu Ala Asp Phe Ala Asp Ala Leu Glu His Pro
1805 1810 1815
Leu Arg Val Pro Lys Pro Asn Thr Ile Glu Leu Ile Ala Met Asp
1820 1825 1830
Leu Pro Met Val Ser Gly Asp Arg Ile His Cys Leu Asp Ile Leu
1835 1840 1845
Phe Ala Phe Thr Lys Arg Val Leu Gly Asp Ser Gly Glu Leu Asp
1850 1855 1860
Ile Leu Arg Gln Gln Met Glu Glu Arg Phe Val Ala Ser Asn Pro
1865 1870 1875
Ser Lys Val Ser Tyr Glu Pro Ile Thr Thr Thr Leu Arg Arg Lys
1880 1885 1890
Gln Glu Glu Val Ser Ala Val Val Leu Gln Arg Ala Tyr Arg Gly
1895 1900 1905
His Leu Ala Arg Arg Gly Phe Ile Cys Lys Lys Thr Thr Ser Asn
1910 1915 1920
Lys Leu Glu Asn Gly Gly Thr His Arg Glu Lys Lys Glu Ser Thr
1925 1930 1935
Pro Ser Thr Ala Ser Leu Pro Ser Tyr Asp Ser Val Thr Lys Pro
1940 1945 1950
Glu Lys Glu Lys Gln Gln Arg Ala Glu Glu Gly Arg Arg Glu Arg
1955 1960 1965
Ala Lys Arg Gln Lys Glu Val Arg Glu Ser Lys Cys
1970 1975 1980
<210> 34
<211> 1988
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(1988)
<223> 人 Nav 1.7（图 9）
<400> 34
Met Ala Met Leu Pro Pro Pro Gly Pro Gln Ser Phe Val His Phe Thr
1 5 10 15
Lys Gln Ser Leu Ala Leu Ile Glu Gln Arg Ile Ala Glu Arg Lys Ser
20 25 30
Lys Glu Pro Lys Glu Glu Lys Lys Asp Asp Asp Glu Glu Ala Pro Lys
35 40 45
Pro Ser Ser Asp Leu Glu Ala Gly Lys Gln Leu Pro Phe Ile Tyr Gly
50 55 60
Asp Ile Pro Pro Gly Met Val Ser Glu Pro Leu Glu Asp Leu Asp Pro
65 70 75 80
Tyr Tyr Ala Asp Lys Lys Thr Phe Ile Val Leu Asn Lys Gly Lys Thr
85 90 95
Ile Phe Arg Phe Asn Ala Thr Pro Ala Leu Tyr Met Leu Ser Pro Phe
100 105 110
Ser Pro Leu Arg Arg Ile Ser Ile Lys Ile Leu Val His Ser Leu Phe
115 120 125
Ser Met Leu Ile Met Cys Thr Ile Leu Thr Asn Cys Ile Phe Met Thr
130 135 140
Met Asn Asn Pro Pro Asp Trp Thr Lys Asn Val Glu Tyr Thr Phe Thr
145 150 155 160
Gly Ile Tyr Thr Phe Glu Ser Leu Val Lys Ile Leu Ala Arg Gly Phe
165 170 175
Cys Val Gly Glu Phe Thr Phe Leu Arg Asp Pro Trp Asn Trp Leu Asp
180 185 190
Phe Val Val Ile Val Phe Ala Tyr Leu Thr Glu Phe Val Asn Leu Gly
195 200 205
Asn Val Ser Ala Leu Arg Thr Phe Arg Val Leu Arg Ala Leu Lys Thr
210 215 220
Ile Ser Val Ile Pro Gly Leu Lys Thr Ile Val Gly Ala Leu Ile Gln
225 230 235 240
Ser Val Lys Lys Leu Ser Asp Val Met Ile Leu Thr Val Phe Cys Leu
245 250 255
Ser Val Phe Ala Leu Ile Gly Leu Gln Leu Phe Met Gly Asn Leu Lys
260 265 270
His Lys Cys Phe Arg Asn Ser Leu Glu Asn Asn Glu Thr Leu Glu Ser
275 280 285
Ile Met Asn Thr Leu Glu Ser Glu Glu Asp Phe Arg Lys Tyr Phe Tyr
290 295 300
Tyr Leu Glu Gly Ser Lys Asp Ala Leu Leu Cys Gly Phe Ser Thr Asp
305 310 315 320
Ser Gly Gln Cys Pro Glu Gly Tyr Thr Cys Val Lys Ile Gly Arg Asn
325 330 335
Pro Asp Tyr Gly Tyr Thr Ser Phe Asp Thr Phe Ser Trp Ala Phe Leu
340 345 350
Ala Leu Phe Arg Leu Met Thr Gln Asp Tyr Trp Glu Asn Leu Tyr Gln
355 360 365
Gln Thr Leu Arg Ala Ala Gly Lys Thr Tyr Met Ile Phe Phe Val Val
370 375 380
Val Ile Phe Leu Gly Ser Phe Tyr Leu Ile Asn Leu Ile Leu Ala Val
385 390 395 400
Val Ala Met Ala Tyr Glu Glu Gln Asn Gln Ala Asn Ile Glu Glu Ala
405 410 415
Lys Gln Lys Glu Leu Glu Phe Gln Gln Met Leu Asp Arg Leu Lys Lys
420 425 430
Glu Gln Glu Glu Ala Glu Ala Ile Ala Ala Ala Ala Ala Glu Tyr Thr
435 440 445
Ser Ile Arg Arg Ser Arg Ile Met Gly Leu Ser Glu Ser Ser Ser Glu
450 455 460
Thr Ser Lys Leu Ser Ser Lys Ser Ala Lys Glu Arg Arg Asn Arg Arg
465 470 475 480
Lys Lys Lys Asn Gln Lys Lys Leu Ser Ser Gly Glu Glu Lys Gly Asp
485 490 495
Ala Glu Lys Leu Ser Lys Ser Glu Ser Glu Asp Ser Ile Arg Arg Lys
500 505 510
Ser Phe His Leu Gly Val Glu Gly His Arg Arg Ala His Glu Lys Arg
515 520 525
Leu Ser Thr Pro Asn Gln Ser Pro Leu Ser Ile Arg Gly Ser Leu Phe
530 535 540
Ser Ala Arg Arg Ser Ser Arg Thr Ser Leu Phe Ser Phe Lys Gly Arg
545 550 555 560
Gly Arg Asp Ile Gly Ser Glu Thr Glu Phe Ala Asp Asp Glu His Ser
565 570 575
Ile Phe Gly Asp Asn Glu Ser Arg Arg Gly Ser Leu Phe Val Pro His
580 585 590
Arg Pro Gln Glu Arg Arg Ser Ser Asn Ile Ser Gln Ala Ser Arg Ser
595 600 605
Pro Pro Met Leu Pro Val Asn Gly Lys Met His Ser Ala Val Asp Cys
610 615 620
Asn Gly Val Val Ser Leu Val Asp Gly Arg Ser Ala Leu Met Leu Pro
625 630 635 640
Asn Gly Gln Leu Leu Pro Glu Val Ile Ile Asp Lys Ala Thr Ser Asp
645 650 655
Asp Ser Gly Thr Thr Asn Gln Ile His Lys Lys Arg Arg Cys Ser Ser
660 665 670
Tyr Leu Leu Ser Glu Asp Met Leu Asn Asp Pro Asn Leu Arg Gln Arg
675 680 685
Ala Met Ser Arg Ala Ser Ile Leu Thr Asn Thr Val Glu Glu Leu Glu
690 695 700
Glu Ser Arg Gln Lys Cys Pro Pro Trp Trp Tyr Arg Phe Ala His Lys
705 710 715 720
Phe Leu Ile Trp Asn Cys Ser Pro Tyr Trp Ile Lys Phe Lys Lys Cys
725 730 735
Ile Tyr Phe Ile Val Met Asp Pro Phe Val Asp Leu Ala Ile Thr Ile
740 745 750
Cys Ile Val Leu Asn Thr Leu Phe Met Ala Met Glu His His Pro Met
755 760 765
Thr Glu Glu Phe Lys Asn Val Leu Ala Ile Gly Asn Leu Val Phe Thr
770 775 780
Gly Ile Phe Ala Ala Glu Met Val Leu Lys Leu Ile Ala Met Asp Pro
785 790 795 800
Tyr Glu Tyr Phe Gln Val Gly Trp Asn Ile Phe Asp Ser Leu Ile Val
805 810 815
Thr Leu Ser Leu Val Glu Leu Phe Leu Ala Asp Val Glu Gly Leu Ser
820 825 830
Val Leu Arg Ser Phe Arg Leu Leu Arg Val Phe Lys Leu Ala Lys Ser
835 840 845
Trp Pro Thr Leu Asn Met Leu Ile Lys Ile Ile Gly Asn Ser Val Gly
850 855 860
Ala Leu Gly Asn Leu Thr Leu Val Leu Ala Ile Ile Val Phe Ile Phe
865 870 875 880
Ala Val Val Gly Met Gln Leu Phe Gly Lys Ser Tyr Lys Glu Cys Val
885 890 895
Cys Lys Ile Asn Asp Asp Cys Thr Leu Pro Arg Trp His Met Asn Asp
900 905 910
Phe Phe His Ser Phe Leu Ile Val Phe Arg Val Leu Cys Gly Glu Trp
915 920 925
Ile Glu Thr Met Trp Asp Cys Met Glu Val Ala Gly Gln Ala Met Cys
930 935 940
Leu Ile Val Tyr Met Met Val Met Val Ile Gly Asn Leu Val Val Leu
945 950 955 960
Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe Ser Ser Asp Asn Leu
965 970 975
Thr Ala Ile Glu Glu Asp Pro Asp Ala Asn Asn Leu Gln Ile Ala Val
980 985 990
Thr Arg Ile Lys Lys Gly Ile Asn Tyr Val Lys Gln Thr Leu Arg Glu
995 1000 1005
Phe Ile Leu Lys Ala Phe Ser Lys Lys Pro Lys Ile Ser Arg Glu
1010 1015 1020
Ile Arg Gln Ala Glu Asp Leu Asn Thr Lys Lys Glu Asn Tyr Ile
1025 1030 1035
Ser Asn His Thr Leu Ala Glu Met Ser Lys Gly His Asn Phe Leu
1040 1045 1050
Lys Glu Lys Asp Lys Ile Ser Gly Phe Gly Ser Ser Val Asp Lys
1055 1060 1065
His Leu Met Glu Asp Ser Asp Gly Gln Ser Phe Ile His Asn Pro
1070 1075 1080
Ser Leu Thr Val Thr Val Pro Ile Ala Pro Gly Glu Ser Asp Leu
1085 1090 1095
Glu Asn Met Asn Ala Glu Glu Leu Ser Ser Asp Ser Asp Ser Glu
1100 1105 1110
Tyr Ser Lys Val Arg Leu Asn Arg Ser Ser Ser Ser Glu Cys Ser
1115 1120 1125
Thr Val Asp Asn Pro Leu Pro Gly Glu Gly Glu Glu Ala Glu Ala
1130 1135 1140
Glu Pro Met Asn Ser Asp Glu Pro Glu Ala Cys Phe Thr Asp Gly
1145 1150 1155
Cys Val Trp Arg Phe Ser Cys Cys Gln Val Asn Ile Glu Ser Gly
1160 1165 1170
Lys Gly Lys Ile Trp Trp Asn Ile Arg Lys Thr Cys Tyr Lys Ile
1175 1180 1185
Val Glu His Ser Trp Phe Glu Ser Phe Ile Val Leu Met Ile Leu
1190 1195 1200
Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Ile Glu Arg
1205 1210 1215
Lys Lys Thr Ile Lys Ile Ile Leu Glu Tyr Ala Asp Lys Ile Phe
1220 1225 1230
Thr Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Trp Ile Ala Tyr
1235 1240 1245
Gly Tyr Lys Thr Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe
1250 1255 1260
Leu Ile Val Asp Val Ser Leu Val Thr Leu Val Ala Asn Thr Leu
1265 1270 1275
Gly Tyr Ser Asp Leu Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg
1280 1285 1290
Ala Leu Arg Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg
1295 1300 1305
Val Val Val Asn Ala Leu Ile Gly Ala Ile Pro Ser Ile Met Asn
1310 1315 1320
Val Leu Leu Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Ile Met
1325 1330 1335
Gly Val Asn Leu Phe Ala Gly Lys Phe Tyr Glu Cys Ile Asn Thr
1340 1345 1350
Thr Asp Gly Ser Arg Phe Pro Ala Ser Gln Val Pro Asn Arg Ser
1355 1360 1365
Glu Cys Phe Ala Leu Met Asn Val Ser Gln Asn Val Arg Trp Lys
1370 1375 1380
Asn Leu Lys Val Asn Phe Asp Asn Val Gly Leu Gly Tyr Leu Ser
1385 1390 1395
Leu Leu Gln Val Ala Thr Phe Lys Gly Trp Thr Ile Ile Met Tyr
1400 1405 1410
Ala Ala Val Asp Ser Val Asn Val Asp Lys Gln Pro Lys Tyr Glu
1415 1420 1425
Tyr Ser Leu Tyr Met Tyr Ile Tyr Phe Val Val Phe Ile Ile Phe
1430 1435 1440
Gly Ser Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp
1445 1450 1455
Asn Phe Asn Gln Gln Lys Lys Lys Leu Gly Gly Gln Asp Ile Phe
1460 1465 1470
Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu
1475 1480 1485
Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg Pro Gly Asn Lys
1490 1495 1500
Ile Gln Gly Cys Ile Phe Asp Leu Val Thr Asn Gln Ala Phe Asp
1505 1510 1515
Ile Ser Ile Met Val Leu Ile Cys Leu Asn Met Val Thr Met Met
1520 1525 1530
Val Glu Lys Glu Gly Gln Ser Gln His Met Thr Glu Val Leu Tyr
1535 1540 1545
Trp Ile Asn Val Val Phe Ile Ile Leu Phe Thr Gly Glu Cys Val
1550 1555 1560
Leu Lys Leu Ile Ser Leu Arg His Tyr Tyr Phe Thr Val Gly Trp
1565 1570 1575
Asn Ile Phe Asp Phe Val Val Val Ile Ile Ser Ile Val Gly Met
1580 1585 1590
Phe Leu Ala Asp Leu Ile Glu Thr Tyr Phe Val Ser Pro Thr Leu
1595 1600 1605
Phe Arg Val Ile Arg Leu Ala Arg Ile Gly Arg Ile Leu Arg Leu
1610 1615 1620
Val Lys Gly Ala Lys Gly Ile Arg Thr Leu Leu Phe Ala Leu Met
1625 1630 1635
Met Ser Leu Pro Ala Leu Phe Asn Ile Gly Leu Leu Leu Phe Leu
1640 1645 1650
Val Met Phe Ile Tyr Ala Ile Phe Gly Met Ser Asn Phe Ala Tyr
1655 1660 1665
Val Lys Lys Glu Asp Gly Ile Asn Asp Met Phe Asn Phe Glu Thr
1670 1675 1680
Phe Gly Asn Ser Met Ile Cys Leu Phe Gln Ile Thr Thr Ser Ala
1685 1690 1695
Gly Trp Asp Gly Leu Leu Ala Pro Ile Leu Asn Ser Lys Pro Pro
1700 1705 1710
Asp Cys Asp Pro Lys Lys Val His Pro Gly Ser Ser Val Glu Gly
1715 1720 1725
Asp Cys Gly Asn Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr
1730 1735 1740
Ile Ile Ile Ser Phe Leu Val Val Val Asn Met Tyr Ile Ala Val
1745 1750 1755
Ile Leu Glu Asn Phe Ser Val Ala Thr Glu Glu Ser Thr Glu Pro
1760 1765 1770
Leu Ser Glu Asp Asp Phe Glu Met Phe Tyr Glu Val Trp Glu Lys
1775 1780 1785
Phe Asp Pro Asp Ala Thr Gln Phe Ile Glu Phe Ser Lys Leu Ser
1790 1795 1800
Asp Phe Ala Ala Ala Leu Asp Pro Pro Leu Leu Ile Ala Lys Pro
1805 1810 1815
Asn Lys Val Gln Leu Ile Ala Met Asp Leu Pro Met Val Ser Gly
1820 1825 1830
Asp Arg Ile His Cys Leu Asp Ile Leu Phe Ala Phe Thr Lys Arg
1835 1840 1845
Val Leu Gly Glu Ser Gly Glu Met Asp Ser Leu Arg Ser Gln Met
1850 1855 1860
Glu Glu Arg Phe Met Ser Ala Asn Pro Ser Lys Val Ser Tyr Glu
1865 1870 1875
Pro Ile Thr Thr Thr Leu Lys Arg Lys Gln Glu Asp Val Ser Ala
1880 1885 1890
Thr Val Ile Gln Arg Ala Tyr Arg Arg Tyr Arg Leu Arg Gln Asn
1895 1900 1905
Val Lys Asn Ile Ser Ser Ile Tyr Ile Lys Asp Gly Asp Arg Asp
1910 1915 1920
Asp Asp Leu Leu Asn Lys Lys Asp Met Ala Phe Asp Asn Val Asn
1925 1930 1935
Glu Asn Ser Ser Pro Glu Lys Thr Asp Ala Thr Ser Ser Thr Thr
1940 1945 1950
Ser Pro Pro Ser Tyr Asp Ser Val Thr Lys Pro Asp Lys Glu Lys
1955 1960 1965
Tyr Glu Gln Asp Arg Thr Glu Lys Glu Asp Lys Gly Lys Asp Ser
1970 1975 1980
Lys Glu Ser Lys Lys
1985
<210> 35
<211> 1956
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(1956)
<223> 人 Nav 1.8（图 9）
<400> 35
Met Glu Phe Pro Ile Gly Ser Leu Glu Thr Asn Asn Phe Arg Arg Phe
1 5 10 15
Thr Pro Glu Ser Leu Val Glu Ile Glu Lys Gln Ile Ala Ala Lys Gln
20 25 30
Gly Thr Lys Lys Ala Arg Glu Lys His Arg Glu Gln Lys Asp Gln Glu
35 40 45
Glu Lys Pro Arg Pro Gln Leu Asp Leu Lys Ala Cys Asn Gln Leu Pro
50 55 60
Lys Phe Tyr Gly Glu Leu Pro Ala Glu Leu Ile Gly Glu Pro Leu Glu
65 70 75 80
Asp Leu Asp Pro Phe Tyr Ser Thr His Arg Thr Phe Met Val Leu Asn
85 90 95
Lys Gly Arg Thr Ile Ser Arg Phe Ser Ala Thr Arg Ala Leu Trp Leu
100 105 110
Phe Ser Pro Phe Asn Leu Ile Arg Arg Thr Ala Ile Lys Val Ser Val
115 120 125
His Ser Trp Phe Ser Leu Phe Ile Thr Val Thr Ile Leu Val Asn Cys
130 135 140
Val Cys Met Thr Arg Thr Asp Leu Pro Glu Lys Ile Glu Tyr Val Phe
145 150 155 160
Thr Val Ile Tyr Thr Phe Glu Ala Leu Ile Lys Ile Leu Ala Arg Gly
165 170 175
Phe Cys Leu Asn Glu Phe Thr Tyr Leu Arg Asp Pro Trp Asn Trp Leu
180 185 190
Asp Phe Ser Val Ile Thr Leu Ala Tyr Val Gly Thr Ala Ile Asp Leu
195 200 205
Arg Gly Ile Ser Gly Leu Arg Thr Phe Arg Val Leu Arg Ala Leu Lys
210 215 220
Thr Val Ser Val Ile Pro Gly Leu Lys Val Ile Val Gly Ala Leu Ile
225 230 235 240
His Ser Val Lys Lys Leu Ala Asp Val Thr Ile Leu Thr Ile Phe Cys
245 250 255
Leu Ser Val Phe Ala Leu Val Gly Leu Gln Leu Phe Lys Gly Asn Leu
260 265 270
Lys Asn Lys Cys Val Lys Asn Asp Met Ala Val Asn Glu Thr Thr Asn
275 280 285
Tyr Ser Ser His Arg Lys Pro Asp Ile Tyr Ile Asn Lys Arg Gly Thr
290 295 300
Ser Asp Pro Leu Leu Cys Gly Asn Gly Ser Asp Ser Gly His Cys Pro
305 310 315 320
Asp Gly Tyr Ile Cys Leu Lys Thr Ser Asp Asn Pro Asp Phe Asn Tyr
325 330 335
Thr Ser Phe Asp Ser Phe Ala Trp Ala Phe Leu Ser Leu Phe Arg Leu
340 345 350
Met Thr Gln Asp Ser Trp Glu Arg Leu Tyr Gln Gln Thr Leu Arg Thr
355 360 365
Ser Gly Lys Ile Tyr Met Ile Phe Phe Val Leu Val Ile Phe Leu Gly
370 375 380
Ser Phe Tyr Leu Val Asn Leu Ile Leu Ala Val Val Thr Met Ala Tyr
385 390 395 400
Glu Glu Gln Asn Gln Ala Thr Thr Asp Glu Ile Glu Ala Lys Glu Lys
405 410 415
Lys Phe Gln Glu Ala Leu Glu Met Leu Arg Lys Glu Gln Glu Val Leu
420 425 430
Ala Ala Leu Gly Ile Asp Thr Thr Ser Leu His Ser His Asn Gly Ser
435 440 445
Pro Leu Thr Ser Lys Asn Ala Ser Glu Arg Arg His Arg Ile Lys Pro
450 455 460
Arg Val Ser Glu Gly Ser Thr Glu Asp Asn Lys Ser Pro Arg Ser Asp
465 470 475 480
Pro Tyr Asn Gln Arg Arg Met Ser Phe Leu Gly Leu Ala Ser Gly Lys
485 490 495
Arg Arg Ala Ser His Gly Ser Val Phe His Phe Arg Ser Pro Gly Arg
500 505 510
Asp Ile Ser Leu Pro Glu Gly Val Thr Asp Asp Gly Val Phe Pro Gly
515 520 525
Asp His Glu Ser His Arg Gly Ser Leu Leu Leu Gly Gly Gly Ala Gly
530 535 540
Gln Gln Gly Pro Leu Pro Arg Ser Pro Leu Pro Gln Pro Ser Asn Pro
545 550 555 560
Asp Ser Arg His Gly Glu Asp Glu His Gln Pro Pro Pro Thr Ser Glu
565 570 575
Leu Ala Pro Gly Ala Val Asp Val Ser Ala Phe Asp Ala Gly Gln Lys
580 585 590
Lys Thr Phe Leu Ser Ala Glu Tyr Leu Asp Glu Pro Phe Arg Ala Gln
595 600 605
Arg Ala Met Ser Val Val Ser Ile Ile Thr Ser Val Leu Glu Glu Leu
610 615 620
Glu Glu Ser Glu Gln Lys Cys Pro Pro Cys Leu Thr Ser Leu Ser Gln
625 630 635 640
Lys Tyr Leu Ile Trp Asp Cys Cys Pro Met Trp Val Lys Leu Lys Thr
645 650 655
Ile Leu Phe Gly Leu Val Thr Asp Pro Phe Ala Glu Leu Thr Ile Thr
660 665 670
Leu Cys Ile Val Val Asn Thr Ile Phe Met Ala Met Glu His His Gly
675 680 685
Met Ser Pro Thr Phe Glu Ala Met Leu Gln Ile Gly Asn Ile Val Phe
690 695 700
Thr Ile Phe Phe Thr Ala Glu Met Val Phe Lys Ile Ile Ala Phe Asp
705 710 715 720
Pro Tyr Tyr Tyr Phe Gln Lys Lys Trp Asn Ile Phe Asp Cys Ile Ile
725 730 735
Val Thr Val Ser Leu Leu Glu Leu Gly Val Ala Lys Lys Gly Ser Leu
740 745 750
Ser Val Leu Arg Ser Phe Arg Leu Leu Arg Val Phe Lys Leu Ala Lys
755 760 765
Ser Trp Pro Thr Leu Asn Thr Leu Ile Lys Ile Ile Gly Asn Ser Val
770 775 780
Gly Ala Leu Gly Asn Leu Thr Ile Ile Leu Ala Ile Ile Val Phe Val
785 790 795 800
Phe Ala Leu Val Gly Lys Gln Leu Leu Gly Glu Asn Tyr Arg Asn Asn
805 810 815
Arg Lys Asn Ile Ser Ala Pro His Glu Asp Trp Pro Arg Trp His Met
820 825 830
His Asp Phe Phe His Ser Phe Leu Ile Val Phe Arg Ile Leu Cys Gly
835 840 845
Glu Trp Ile Glu Asn Met Trp Ala Cys Met Glu Val Gly Gln Lys Ser
850 855 860
Ile Cys Leu Ile Leu Phe Leu Thr Val Met Val Leu Gly Asn Leu Val
865 870 875 880
Val Leu Asn Leu Phe Ile Ala Leu Leu Leu Asn Ser Phe Ser Ala Asp
885 890 895
Asn Leu Thr Ala Pro Glu Asp Asp Gly Glu Val Asn Asn Leu Gln Val
900 905 910
Ala Leu Ala Arg Ile Gln Val Phe Gly His Arg Thr Lys Gln Ala Leu
915 920 925
Cys Ser Phe Phe Ser Arg Ser Cys Pro Phe Pro Gln Pro Lys Ala Glu
930 935 940
Pro Glu Leu Val Val Lys Leu Pro Leu Ser Ser Ser Lys Ala Glu Asn
945 950 955 960
His Ile Ala Ala Asn Thr Ala Arg Gly Ser Ser Gly Gly Leu Gln Ala
965 970 975
Pro Arg Gly Pro Arg Asp Glu His Ser Asp Phe Ile Ala Asn Pro Thr
980 985 990
Val Trp Val Ser Val Pro Ile Ala Glu Gly Glu Ser Asp Leu Asp Asp
995 1000 1005
Leu Glu Asp Asp Gly Gly Glu Asp Ala Gln Ser Phe Gln Gln Glu
1010 1015 1020
Val Ile Pro Lys Gly Gln Gln Glu Gln Leu Gln Gln Val Glu Arg
1025 1030 1035
Cys Gly Asp His Leu Thr Pro Arg Ser Pro Gly Thr Gly Thr Ser
1040 1045 1050
Ser Glu Asp Leu Ala Pro Ser Leu Gly Glu Thr Trp Lys Asp Glu
1055 1060 1065
Ser Val Pro Gln Val Pro Ala Glu Gly Val Asp Asp Thr Ser Ser
1070 1075 1080
Ser Glu Gly Ser Thr Val Asp Cys Leu Asp Pro Glu Glu Ile Leu
1085 1090 1095
Arg Lys Ile Pro Glu Leu Ala Asp Asp Leu Glu Glu Pro Asp Asp
1100 1105 1110
Cys Phe Thr Glu Gly Cys Ile Arg His Cys Pro Cys Cys Lys Leu
1115 1120 1125
Asp Thr Thr Lys Ser Pro Trp Asp Val Gly Trp Gln Val Arg Lys
1130 1135 1140
Thr Cys Tyr Arg Ile Val Glu His Ser Trp Phe Glu Ser Phe Ile
1145 1150 1155
Ile Phe Met Ile Leu Leu Ser Ser Gly Ser Leu Ala Phe Glu Asp
1160 1165 1170
Tyr Tyr Leu Asp Gln Lys Pro Thr Val Lys Ala Leu Leu Glu Tyr
1175 1180 1185
Thr Asp Arg Val Phe Thr Phe Ile Phe Val Phe Glu Met Leu Leu
1190 1195 1200
Lys Trp Val Ala Tyr Gly Phe Lys Lys Tyr Phe Thr Asn Ala Trp
1205 1210 1215
Cys Trp Leu Asp Phe Leu Ile Val Asn Ile Ser Leu Ile Ser Leu
1220 1225 1230
Thr Ala Lys Ile Leu Glu Tyr Ser Glu Val Ala Pro Ile Lys Ala
1235 1240 1245
Leu Arg Thr Leu Arg Ala Leu Arg Pro Leu Arg Ala Leu Ser Arg
1250 1255 1260
Phe Glu Gly Met Arg Val Val Val Asp Ala Leu Val Gly Ala Ile
1265 1270 1275
Pro Ser Ile Met Asn Val Leu Leu Val Cys Leu Ile Phe Trp Leu
1280 1285 1290
Ile Phe Ser Ile Met Gly Val Asn Leu Phe Ala Gly Lys Phe Trp
1295 1300 1305
Arg Cys Ile Asn Tyr Thr Asp Gly Glu Phe Ser Leu Val Pro Leu
1310 1315 1320
Ser Ile Val Asn Asn Lys Ser Asp Cys Lys Ile Gln Asn Ser Thr
1325 1330 1335
Gly Ser Phe Phe Trp Val Asn Val Lys Val Asn Phe Asp Asn Val
1340 1345 1350
Ala Met Gly Tyr Leu Ala Leu Leu Gln Val Ala Thr Phe Lys Gly
1355 1360 1365
Trp Met Asp Ile Met Tyr Ala Ala Val Asp Ser Arg Glu Val Asn
1370 1375 1380
Met Gln Pro Lys Trp Glu Asp Asn Val Tyr Met Tyr Leu Tyr Phe
1385 1390 1395
Val Ile Phe Ile Ile Phe Gly Gly Phe Phe Thr Leu Asn Leu Phe
1400 1405 1410
Val Gly Val Ile Ile Asp Asn Phe Asn Gln Gln Lys Lys Lys Leu
1415 1420 1425
Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr
1430 1435 1440
Asn Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile
1445 1450 1455
Pro Arg Pro Leu Asn Lys Phe Gln Gly Phe Val Phe Asp Ile Val
1460 1465 1470
Thr Arg Gln Ala Phe Asp Ile Thr Ile Met Val Leu Ile Cys Leu
1475 1480 1485
Asn Met Ile Thr Met Met Val Glu Thr Asp Asp Gln Ser Glu Glu
1490 1495 1500
Lys Thr Lys Ile Leu Gly Lys Ile Asn Gln Phe Phe Val Ala Val
1505 1510 1515
Phe Thr Gly Glu Cys Val Met Lys Met Phe Ala Leu Arg Gln Tyr
1520 1525 1530
Tyr Phe Thr Asn Gly Trp Asn Val Phe Asp Phe Ile Val Val Val
1535 1540 1545
Leu Ser Ile Ala Ser Leu Ile Phe Ser Ala Ile Leu Lys Ser Leu
1550 1555 1560
Gln Ser Tyr Phe Ser Pro Thr Leu Phe Arg Val Ile Arg Leu Ala
1565 1570 1575
Arg Ile Gly Arg Ile Leu Arg Leu Ile Arg Ala Ala Lys Gly Ile
1580 1585 1590
Arg Thr Leu Leu Phe Ala Leu Met Met Ser Leu Pro Ala Leu Phe
1595 1600 1605
Asn Ile Gly Leu Leu Leu Phe Leu Val Met Phe Ile Tyr Ser Ile
1610 1615 1620
Phe Gly Met Ser Ser Phe Pro His Val Arg Trp Glu Ala Gly Ile
1625 1630 1635
Asp Asp Met Phe Asn Phe Gln Thr Phe Ala Asn Ser Met Leu Cys
1640 1645 1650
Leu Phe Gln Ile Thr Thr Ser Ala Gly Trp Asp Gly Leu Leu Ser
1655 1660 1665
Pro Ile Leu Asn Thr Gly Pro Pro Tyr Cys Asp Pro Asn Leu Pro
1670 1675 1680
Asn Ser Asn Gly Thr Arg Gly Asp Cys Gly Ser Pro Ala Val Gly
1685 1690 1695
Ile Ile Phe Phe Thr Thr Tyr Ile Ile Ile Ser Phe Leu Ile Met
1700 1705 1710
Val Asn Met Tyr Ile Ala Val Ile Leu Glu Asn Phe Asn Val Ala
1715 1720 1725
Thr Glu Glu Ser Thr Glu Pro Leu Ser Glu Asp Asp Phe Asp Met
1730 1735 1740
Phe Tyr Glu Thr Trp Glu Lys Phe Asp Pro Glu Ala Thr Gln Phe
1745 1750 1755
Ile Thr Phe Ser Ala Leu Ser Asp Phe Ala Asp Thr Leu Ser Gly
1760 1765 1770
Pro Leu Arg Ile Pro Lys Pro Asn Arg Asn Ile Leu Ile Gln Met
1775 1780 1785
Asp Leu Pro Leu Val Pro Gly Asp Lys Ile His Cys Leu Asp Ile
1790 1795 1800
Leu Phe Ala Phe Thr Lys Asn Val Leu Gly Glu Ser Gly Glu Leu
1805 1810 1815
Asp Ser Leu Lys Ala Asn Met Glu Glu Lys Phe Met Ala Thr Asn
1820 1825 1830
Leu Ser Lys Ser Ser Tyr Glu Pro Ile Ala Thr Thr Leu Arg Trp
1835 1840 1845
Lys Gln Glu Asp Ile Ser Ala Thr Val Ile Gln Lys Ala Tyr Arg
1850 1855 1860
Ser Tyr Val Leu His Arg Ser Met Ala Leu Ser Asn Thr Pro Cys
1865 1870 1875
Val Pro Arg Ala Glu Glu Glu Ala Ala Ser Leu Pro Asp Glu Gly
1880 1885 1890
Phe Val Ala Phe Thr Ala Asn Glu Asn Cys Val Leu Pro Asp Lys
1895 1900 1905
Ser Glu Thr Ala Ser Ala Thr Ser Phe Pro Pro Ser Tyr Glu Ser
1910 1915 1920
Val Thr Arg Gly Leu Ser Asp Arg Val Asn Met Arg Thr Ser Ser
1925 1930 1935
Ser Ile Gln Asn Glu Asp Glu Ala Thr Ser Met Glu Leu Ile Ala
1940 1945 1950
Pro Gly Pro
1955
<210> 36
<211> 1791
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(1791)
<223> 人 Nav 1.9（图 9）
<400> 36
Met Asp Asp Arg Cys Tyr Pro Val Ile Phe Pro Asp Glu Arg Asn Phe
1 5 10 15
Arg Pro Phe Thr Ser Asp Ser Leu Ala Ala Ile Glu Lys Arg Ile Ala
20 25 30
Ile Gln Lys Glu Lys Lys Lys Ser Lys Asp Gln Thr Gly Glu Val Pro
35 40 45
Gln Pro Arg Pro Gln Leu Asp Leu Lys Ala Ser Arg Lys Leu Pro Lys
50 55 60
Leu Tyr Gly Asp Ile Pro Arg Glu Leu Ile Gly Lys Pro Leu Glu Asp
65 70 75 80
Leu Asp Pro Phe Tyr Arg Asn His Lys Thr Phe Met Val Leu Asn Arg
85 90 95
Lys Arg Thr Ile Tyr Arg Phe Ser Ala Lys His Ala Leu Phe Ile Phe
100 105 110
Gly Pro Phe Asn Ser Ile Arg Ser Leu Ala Ile Arg Val Ser Val His
115 120 125
Ser Leu Phe Ser Met Phe Ile Ile Gly Thr Val Ile Ile Asn Cys Val
130 135 140
Phe Met Ala Thr Gly Pro Ala Lys Asn Ser Asn Ser Asn Asn Thr Asp
145 150 155 160
Ile Ala Glu Cys Val Phe Thr Gly Ile Tyr Ile Phe Glu Ala Leu Ile
165 170 175
Lys Ile Leu Ala Arg Gly Phe Ile Leu Asp Glu Phe Ser Phe Leu Arg
180 185 190
Asp Pro Trp Asn Trp Leu Asp Ser Ile Val Ile Gly Ile Ala Ile Val
195 200 205
Ser Tyr Ile Pro Gly Ile Thr Ile Lys Leu Leu Pro Leu Arg Thr Phe
210 215 220
Arg Val Phe Arg Ala Leu Lys Ala Ile Ser Val Val Ser Arg Leu Lys
225 230 235 240
Val Ile Val Gly Ala Leu Leu Arg Ser Val Lys Lys Leu Val Asn Val
245 250 255
Ile Ile Leu Thr Phe Phe Cys Leu Ser Ile Phe Ala Leu Val Gly Gln
260 265 270
Gln Leu Phe Met Gly Ser Leu Asn Leu Lys Cys Ile Ser Arg Asp Cys
275 280 285
Lys Asn Ile Ser Asn Pro Glu Ala Tyr Asp His Cys Phe Glu Lys Lys
290 295 300
Glu Asn Ser Pro Glu Phe Lys Met Cys Gly Ile Trp Met Gly Asn Ser
305 310 315 320
Ala Cys Ser Ile Gln Tyr Glu Cys Lys His Thr Lys Ile Asn Pro Asp
325 330 335
Tyr Asn Tyr Thr Asn Phe Asp Asn Phe Gly Trp Ser Phe Leu Ala Met
340 345 350
Phe Arg Leu Met Thr Gln Asp Ser Trp Glu Lys Leu Tyr Gln Gln Thr
355 360 365
Leu Arg Thr Thr Gly Leu Tyr Ser Val Phe Phe Phe Ile Val Val Ile
370 375 380
Phe Leu Gly Ser Phe Tyr Leu Ile Asn Leu Thr Leu Ala Val Val Thr
385 390 395 400
Met Ala Tyr Glu Glu Gln Asn Lys Asn Val Ala Ala Glu Ile Glu Ala
405 410 415
Lys Glu Lys Met Phe Gln Glu Ala Gln Gln Leu Leu Lys Glu Glu Lys
420 425 430
Glu Ala Leu Val Ala Met Gly Ile Asp Arg Ser Ser Leu Thr Ser Leu
435 440 445
Glu Thr Ser Tyr Phe Thr Pro Lys Lys Arg Lys Leu Phe Gly Asn Lys
450 455 460
Lys Arg Lys Ser Phe Phe Leu Arg Glu Ser Gly Lys Asp Gln Pro Pro
465 470 475 480
Gly Ser Asp Ser Asp Glu Asp Cys Gln Lys Lys Pro Gln Leu Leu Glu
485 490 495
Gln Thr Lys Arg Leu Ser Gln Asn Leu Ser Leu Asp His Phe Asp Glu
500 505 510
His Gly Asp Pro Leu Gln Arg Gln Arg Ala Leu Ser Ala Val Ser Ile
515 520 525
Leu Thr Ile Thr Met Lys Glu Gln Glu Lys Ser Gln Glu Pro Cys Leu
530 535 540
Pro Cys Gly Glu Asn Leu Ala Ser Lys Tyr Leu Val Trp Asn Cys Cys
545 550 555 560
Pro Gln Trp Leu Cys Val Lys Lys Val Leu Arg Thr Val Met Thr Asp
565 570 575
Pro Phe Thr Glu Leu Ala Ile Thr Ile Cys Ile Ile Ile Asn Thr Val
580 585 590
Phe Leu Ala Met Glu His His Lys Met Glu Ala Ser Phe Glu Lys Met
595 600 605
Leu Asn Ile Gly Asn Leu Val Phe Thr Ser Ile Phe Ile Ala Glu Met
610 615 620
Cys Leu Lys Ile Ile Ala Leu Asp Pro Tyr His Tyr Phe Arg Arg Gly
625 630 635 640
Trp Asn Ile Phe Asp Ser Ile Val Ala Leu Leu Ser Phe Ala Asp Val
645 650 655
Met Asn Cys Val Leu Gln Lys Arg Ser Trp Pro Phe Leu Arg Ser Phe
660 665 670
Arg Val Leu Arg Val Phe Lys Leu Ala Lys Ser Trp Pro Thr Leu Asn
675 680 685
Thr Leu Ile Lys Ile Ile Gly Asn Ser Val Gly Ala Leu Gly Ser Leu
690 695 700
Thr Val Val Leu Val Ile Val Ile Phe Ile Phe Ser Val Val Gly Met
705 710 715 720
Gln Leu Phe Gly Arg Ser Phe Asn Ser Gln Lys Ser Pro Lys Leu Cys
725 730 735
Asn Pro Thr Gly Pro Thr Val Ser Cys Leu Arg His Trp His Met Gly
740 745 750
Asp Phe Trp His Ser Phe Leu Val Val Phe Arg Ile Leu Cys Gly Glu
755 760 765
Trp Ile Glu Asn Met Trp Glu Cys Met Gln Glu Ala Asn Ala Ser Ser
770 775 780
Ser Leu Cys Val Ile Val Phe Ile Leu Ile Thr Val Ile Gly Lys Leu
785 790 795 800
Val Val Leu Asn Leu Phe Ile Ala Leu Leu Leu Asn Ser Phe Ser Asn
805 810 815
Glu Glu Arg Asn Gly Asn Leu Glu Gly Glu Ala Arg Lys Thr Lys Val
820 825 830
Gln Leu Ala Leu Asp Arg Phe Arg Arg Ala Phe Cys Phe Val Arg His
835 840 845
Thr Leu Glu His Phe Cys His Lys Trp Cys Arg Lys Gln Asn Leu Pro
850 855 860
Gln Gln Lys Glu Val Ala Gly Gly Cys Ala Ala Gln Ser Lys Asp Ile
865 870 875 880
Ile Pro Leu Val Met Glu Met Lys Arg Gly Ser Glu Thr Gln Glu Glu
885 890 895
Leu Gly Ile Leu Thr Ser Val Pro Lys Thr Leu Gly Val Arg His Asp
900 905 910
Trp Thr Trp Leu Ala Pro Leu Ala Glu Glu Glu Asp Asp Val Glu Phe
915 920 925
Ser Gly Glu Asp Asn Ala Gln Arg Ile Thr Gln Pro Glu Pro Glu Gln
930 935 940
Gln Ala Tyr Glu Leu His Gln Glu Asn Lys Lys Pro Thr Ser Gln Arg
945 950 955 960
Val Gln Ser Val Glu Ile Asp Met Phe Ser Glu Asp Glu Pro His Leu
965 970 975
Thr Ile Gln Asp Pro Arg Lys Lys Ser Asp Val Thr Ser Ile Leu Ser
980 985 990
Glu Cys Ser Thr Ile Asp Leu Gln Asp Gly Phe Gly Trp Leu Pro Glu
995 1000 1005
Met Val Pro Lys Lys Gln Pro Glu Arg Cys Leu Pro Lys Gly Phe
1010 1015 1020
Gly Cys Cys Phe Pro Cys Cys Ser Val Asp Lys Arg Lys Pro Pro
1025 1030 1035
Trp Val Ile Trp Trp Asn Leu Arg Lys Thr Cys Tyr Gln Ile Val
1040 1045 1050
Lys His Ser Trp Phe Glu Ser Phe Ile Ile Phe Val Ile Leu Leu
1055 1060 1065
Ser Ser Gly Ala Leu Ile Phe Glu Asp Val His Leu Glu Asn Gln
1070 1075 1080
Pro Lys Ile Gln Glu Leu Leu Asn Cys Thr Asp Ile Ile Phe Thr
1085 1090 1095
His Ile Phe Ile Leu Glu Met Val Leu Lys Trp Val Ala Phe Gly
1100 1105 1110
Phe Gly Lys Tyr Phe Thr Ser Ala Trp Cys Cys Leu Asp Phe Ile
1115 1120 1125
Ile Val Ile Val Ser Val Thr Thr Leu Ile Asn Leu Met Glu Leu
1130 1135 1140
Lys Ser Phe Arg Thr Leu Arg Ala Leu Arg Pro Leu Arg Ala Leu
1145 1150 1155
Ser Gln Phe Glu Gly Met Lys Val Val Val Asn Ala Leu Ile Gly
1160 1165 1170
Ala Ile Pro Ala Ile Leu Asn Val Leu Leu Val Cys Leu Ile Phe
1175 1180 1185
Trp Leu Val Phe Cys Ile Leu Gly Val Tyr Phe Phe Ser Gly Lys
1190 1195 1200
Phe Gly Lys Cys Ile Asn Gly Thr Asp Ser Val Ile Asn Tyr Thr
1205 1210 1215
Ile Ile Thr Asn Lys Ser Gln Cys Glu Ser Gly Asn Phe Ser Trp
1220 1225 1230
Ile Asn Gln Lys Val Asn Phe Asp Asn Val Gly Asn Ala Tyr Leu
1235 1240 1245
Ala Leu Leu Gln Val Ala Thr Phe Lys Gly Trp Met Asp Ile Ile
1250 1255 1260
Tyr Ala Ala Val Asp Ser Thr Glu Lys Glu Gln Gln Pro Glu Phe
1265 1270 1275
Glu Ser Asn Ser Leu Gly Tyr Ile Tyr Phe Val Val Phe Ile Ile
1280 1285 1290
Phe Gly Ser Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile
1295 1300 1305
Asp Asn Phe Asn Gln Gln Gln Lys Lys Leu Gly Gly Gln Asp Ile
1310 1315 1320
Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys
1325 1330 1335
Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg Pro Leu Asn
1340 1345 1350
Lys Cys Gln Gly Leu Val Phe Asp Ile Val Thr Ser Gln Ile Phe
1355 1360 1365
Asp Ile Ile Ile Ile Ser Leu Ile Ile Leu Asn Met Ile Ser Met
1370 1375 1380
Met Ala Glu Ser Tyr Asn Gln Pro Lys Ala Met Lys Ser Ile Leu
1385 1390 1395
Asp His Leu Asn Trp Val Phe Val Val Ile Phe Thr Leu Glu Cys
1400 1405 1410
Leu Ile Lys Ile Phe Ala Leu Arg Gln Tyr Tyr Phe Thr Asn Gly
1415 1420 1425
Trp Asn Leu Phe Asp Cys Val Val Val Leu Leu Ser Ile Val Ser
1430 1435 1440
Thr Met Ile Ser Thr Leu Glu Asn Gln Glu His Ile Pro Phe Pro
1445 1450 1455
Pro Thr Leu Phe Arg Ile Val Arg Leu Ala Arg Ile Gly Arg Ile
1460 1465 1470
Leu Arg Leu Val Arg Ala Ala Arg Gly Ile Arg Thr Leu Leu Phe
1475 1480 1485
Ala Leu Met Met Ser Leu Pro Ser Leu Phe Asn Ile Gly Leu Leu
1490 1495 1500
Leu Phe Leu Ile Met Phe Ile Tyr Ala Ile Leu Gly Met Asn Trp
1505 1510 1515
Phe Ser Lys Val Asn Pro Glu Ser Gly Ile Asp Asp Ile Phe Asn
1520 1525 1530
Phe Lys Thr Phe Ala Ser Ser Met Leu Cys Leu Phe Gln Ile Ser
1535 1540 1545
Thr Ser Ala Gly Trp Asp Ser Leu Leu Ser Pro Met Leu Arg Ser
1550 1555 1560
Lys Glu Ser Cys Asn Ser Ser Ser Glu Asn Cys His Leu Pro Gly
1565 1570 1575
Ile Ala Thr Ser Tyr Phe Val Ser Tyr Ile Ile Ile Ser Phe Leu
1580 1585 1590
Ile Val Val Asn Met Tyr Ile Ala Val Ile Leu Glu Asn Phe Asn
1595 1600 1605
Thr Ala Thr Glu Glu Ser Glu Asp Pro Leu Gly Glu Asp Asp Phe
1610 1615 1620
Asp Ile Phe Tyr Glu Val Trp Glu Lys Phe Asp Pro Glu Ala Thr
1625 1630 1635
Gln Phe Ile Lys Tyr Ser Ala Leu Ser Asp Phe Ala Asp Ala Leu
1640 1645 1650
Pro Glu Pro Leu Arg Val Ala Lys Pro Asn Lys Tyr Gln Phe Leu
1655 1660 1665
Val Met Asp Leu Pro Met Val Ser Glu Asp Arg Leu His Cys Met
1670 1675 1680
Asp Ile Leu Phe Ala Phe Thr Ala Arg Val Leu Gly Gly Ser Asp
1685 1690 1695
Gly Leu Asp Ser Met Lys Ala Met Met Glu Glu Lys Phe Met Glu
1700 1705 1710
Ala Asn Pro Leu Lys Lys Leu Tyr Glu Pro Ile Val Thr Thr Thr
1715 1720 1725
Lys Arg Lys Glu Glu Glu Arg Gly Ala Ala Ile Ile Gln Lys Ala
1730 1735 1740
Phe Arg Lys Tyr Met Met Lys Val Thr Lys Gly Asp Gln Gly Asp
1745 1750 1755
Gln Asn Asp Leu Glu Asn Gly Pro His Ser Pro Leu Gln Thr Leu
1760 1765 1770
Cys Asn Gly Asp Leu Ser Ser Phe Gly Val Ala Lys Gly Lys Val
1775 1780 1785
His Cys Asp
1790
<210> 37
<211> 36
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(36)
<223> 人 Nax 1204-1239（图 14B）
<400> 37
Gly Gly Ser Asn Ile Phe Ile Thr Val Lys Gln Arg Lys Gln Tyr Arg
1 5 10 15
Arg Leu Lys Lys Leu Met Tyr Glu Asp Ser Gln Arg Pro Val Pro Arg
20 25 30
Pro Leu Asn Lys
35
<210> 38
<211> 36
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(36)
<223> 人 Nav 1.1 1494-1529（图 14B）
<400> 38
Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn
1 5 10 15
Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg
20 25 30
Pro Gly Asn Lys
35
<210> 39
<211> 36
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(36)
<223> 人 Nav 1.2 1484-1519（图 14B）
<400> 39
Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn
1 5 10 15
Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg
20 25 30
Pro Ala Asn Lys
35
<210> 40
<211> 36
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(36)
<223> 人 Nav 1.3 1479-1514（图 14B）
<400> 40
Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn
1 5 10 15
Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg
20 25 30
Pro Ala Asn Lys
35
<210> 41
<211> 36
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(36)
<223> 人 Nav 1.4 1206-1241（图 14B）
<400> 41
Gly Gly Lys Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn
1 5 10 15
Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg
20 25 30
Pro Gln Asn Lys
35
<210> 42
<211> 36
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(36)
<223> 人 Nav 1.5 1481-1516（图 14B）
<400> 42
Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn
1 5 10 15
Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg
20 25 30
Pro Leu Asn Lys
35
<210> 43
<211> 36
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(36)
<223> 人 Nav 1.6 1475-1510（图 14B）
<400> 43
Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn
1 5 10 15
Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg
20 25 30
Pro Leu Asn Lys
35
<210> 44
<211> 36
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(36)
<223> 人 Nav 1.7 1468-1503（图 14B）
<400> 44
Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn
1 5 10 15
Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg
20 25 30
Pro Gly Asn Lys
35
<210> 45
<211> 36
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(36)
<223> 人 Nav 1.8 1429-1464（图 14B）
<400> 45
Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn
1 5 10 15
Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg
20 25 30
Pro Leu Asn Lys
35
<210> 46
<211> 36
<212> PRT
<213> 智人
<220>
<221> misc_特征
<222> (1)..(36)
<223> 人 Nav 1.9 1319-1354（图 14B）
<400> 46
Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn
1 5 10 15
Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg
20 25 30
Pro Leu Asn Lys
35
<210> 47
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaX_D1_S6（图 20）
<400> 47
Gly Lys Val Tyr Met Ile Phe Phe Val Val Val Ser Phe Leu Phe Ser
1 5 10 15
Phe Tyr Met Ala Ser Leu Phe Leu Gly Ile Leu Ala Met Ala Tyr
20 25 30
<210> 48
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.1_D1_S6（图 20）
<400> 48
Gly Lys Thr Tyr Met Ile Phe Phe Val Leu Val Ile Phe Leu Gly Ser
1 5 10 15
Phe Tyr Leu Ile Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr
20 25 30
<210> 49
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.2_D1_S6（图 20）
<400> 49
Gly Lys Thr Tyr Met Ile Phe Phe Val Leu Val Ile Phe Leu Gly Ser
1 5 10 15
Phe Tyr Leu Ile Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr
20 25 30
<210> 50
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.3_D1_S6（图 20）
<400> 50
Gly Lys Thr Tyr Met Ile Phe Phe Val Leu Val Ile Phe Leu Gly Ser
1 5 10 15
Phe Tyr Leu Val Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr
20 25 30
<210> 51
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.4_D1_S6（图 20）
<400> 51
Gly Lys Thr Tyr Met Ile Phe Phe Val Val Ile Ile Phe Leu Gly Ser
1 5 10 15
Phe Tyr Leu Ile Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr
20 25 30
<210> 52
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.5_D1_S6（图 20）
<400> 52
Gly Lys Ile Tyr Met Ile Phe Phe Met Leu Val Ile Phe Leu Gly Ser
1 5 10 15
Phe Tyr Leu Val Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr
20 25 30
<210> 53
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.6_D1_S6（图 20）
<400> 53
Gly Lys Thr Tyr Met Ile Phe Phe Val Leu Val Ile Phe Val Gly Ser
1 5 10 15
Phe Tyr Leu Val Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr
20 25 30
<210> 54
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.7_D1_S6（图 20）
<400> 54
Gly Lys Thr Tyr Met Ile Phe Phe Val Val Val Ile Phe Leu Gly Ser
1 5 10 15
Phe Tyr Leu Ile Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr
20 25 30
<210> 55
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.8_D1_S6（图 20）
<400> 55
Gly Lys Ile Tyr Met Ile Phe Phe Val Leu Val Ile Phe Leu Gly Ser
1 5 10 15
Phe Tyr Leu Val Asn Leu Ile Leu Ala Val Val Thr Met Ala Tyr
20 25 30
<210> 56
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.9_D1_S6（图 20）
<400> 56
Gly Leu Tyr Ser Val Phe Phe Phe Ile Val Val Ile Phe Leu Gly Ser
1 5 10 15
Phe Tyr Leu Ile Asn Leu Thr Leu Ala Val Val Thr Met Ala Tyr
20 25 30
<210> 57
<211> 30
<212> PRT
<213> 未知
<220>
<223> NaX_D2_S6（图 20）
<400> 57
Gln Ser Trp Cys Ile Pro Phe Tyr Leu Met Val Ile Leu Ile Gly Asn
1 5 10 15
Leu Leu Val Leu Tyr Leu Phe Leu Ala Leu Val Ser Ser Phe
20 25 30
<210> 58
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.1_D2_S6（图 20）
<400> 58
Gln Ala Met Cys Leu Thr Val Phe Met Met Val Met Val Ile Gly Asn
1 5 10 15
Leu Val Val Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe
20 25 30
<210> 59
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.2_D2_S6（图 20）
<400> 59
Gln Thr Met Cys Leu Thr Val Phe Met Met Val Met Val Ile Gly Asn
1 5 10 15
Leu Val Val Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe
20 25 30
<210> 60
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.3_D2_S6（图 20）
<400> 60
Gln Thr Met Cys Leu Ile Val Phe Met Leu Val Met Val Ile Gly Asn
1 5 10 15
Leu Val Val Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe
20 25 30
<210> 61
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.4_D2_S6（图 20）
<400> 61
Gln Ala Met Cys Leu Thr Val Phe Leu Met Val Met Val Ile Gly Asn
1 5 10 15
Leu Val Val Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe
20 25 30
<210> 62
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.5_D2_S6（图 20）
<400> 62
Gln Ser Leu Cys Leu Leu Val Phe Leu Leu Val Met Val Ile Gly Asn
1 5 10 15
Leu Val Val Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe
20 25 30
<210> 63
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.6_D2_S6（图 20）
<400> 63
Gln Ala Met Cys Leu Ile Val Phe Met Met Val Met Val Ile Gly Asn
1 5 10 15
Leu Val Val Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe
20 25 30
<210> 64
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.7_D2_S6（图 20）
<400> 64
Gln Ala Met Cys Leu Ile Val Tyr Met Met Val Met Val Ile Gly Asn
1 5 10 15
Leu Val Val Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe
20 25 30
<210> 65
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.8_D2_S6（图 20）
<400> 65
Lys Ser Ile Cys Leu Ile Leu Phe Leu Thr Val Met Val Leu Gly Asn
1 5 10 15
Leu Val Val Leu Asn Leu Phe Ile Ala Leu Leu Leu Asn Ser Phe
20 25 30
<210> 66
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.9_D2_S6（图 20）
<400> 66
Ser Ser Leu Cys Val Ile Val Phe Ile Leu Ile Thr Val Ile Gly Lys
1 5 10 15
Leu Val Val Leu Asn Leu Phe Ile Ala Leu Leu Leu Asn Ser Phe
20 25 30
<210> 67
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaX_D3_S6（图 20）
<400> 67
Asn Ile Tyr Met Tyr Cys Tyr Phe Ile Asn Phe Ile Ile Phe Gly Val
1 5 10 15
Phe Leu Pro Leu Ser Met Leu Ile Thr Val Ile Ile Asp Asn Phe
20 25 30
<210> 68
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.1_D3_S6（图 20）
<400> 68
Ser Leu Tyr Met Tyr Leu Tyr Phe Val Ile Phe Ile Ile Phe Gly Ser
1 5 10 15
Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe
20 25 30
<210> 69
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.2_D3_S6（图 20）
<400> 69
Asn Leu Tyr Met Tyr Leu Tyr Phe Val Ile Phe Ile Ile Phe Gly Ser
1 5 10 15
Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe
20 25 30
<210> 70
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.3_D3_S6（图 20）
<400> 70
Asn Leu Tyr Met Tyr Leu Tyr Phe Val Ile Phe Ile Ile Phe Gly Ser
1 5 10 15
Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe
20 25 30
<210> 71
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.4_D3_S6（图 20）
<400> 71
Asn Leu Tyr Met Tyr Leu Tyr Phe Val Ile Phe Ile Ile Phe Gly Ser
1 5 10 15
Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe
20 25 30
<210> 72
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.5_D3_S6（图 20）
<400> 72
Asn Leu Tyr Met Tyr Ile Tyr Phe Val Ile Phe Ile Ile Phe Gly Ser
1 5 10 15
Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe
20 25 30
<210> 73
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.6_D3_S6（图 20）
<400> 73
Asn Ile Tyr Met Tyr Ile Tyr Phe Val Ile Phe Ile Ile Phe Gly Ser
1 5 10 15
Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe
20 25 30
<210> 74
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.7_D3_S6（图 20）
<400> 74
Ser Leu Tyr Met Tyr Ile Tyr Phe Val Val Phe Ile Ile Phe Gly Ser
1 5 10 15
Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe
20 25 30
<210> 75
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.8_D3_S6（图 20）
<400> 75
Asn Val Tyr Met Tyr Leu Tyr Phe Val Ile Phe Ile Ile Phe Gly Gly
1 5 10 15
Phe Phe Thr Leu Asn Leu Phe Val Gly Val Ile Ile Asp Asn Phe
20 25 30
<210> 76
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.9_D3_S6（图 20）
<400> 76
Asn Ser Leu Gly Tyr Ile Tyr Phe Val Val Phe Ile Ile Phe Gly Ser
1 5 10 15
Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe
20 25 30
<210> 77
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaX_D4_S6（图 20）
<400> 77
Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Leu Ile Ser Trp
1 5 10 15
Leu Ile Ile Val Asn Met Tyr Ile Val Val Val Met Glu Phe Leu
20 25 30
<210> 78
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.1_D4_S6（图 20）
<400> 78
Pro Ser Val Gly Ile Phe Phe Phe Val Ser Tyr Ile Ile Ile Ser Phe
1 5 10 15
Leu Val Val Val Asn Met Tyr Ile Ala Val Ile Leu Glu Asn Phe
20 25 30
<210> 79
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.2_D4_S6（图 20）
<400> 79
Pro Ser Val Gly Ile Phe Phe Phe Val Ser Tyr Ile Ile Ile Ser Phe
1 5 10 15
Leu Val Val Val Asn Met Tyr Ile Ala Val Ile Leu Glu Asn Phe
20 25 30
<210> 80
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.3_D4_S6（图 20）
<400> 80
Pro Ser Val Gly Ile Phe Phe Phe Val Ser Tyr Ile Ile Ile Ser Phe
1 5 10 15
Leu Val Val Val Asn Met Tyr Ile Ala Val Ile Leu Glu Asn Phe
20 25 30
<210> 81
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.4_D4_S6（图 20）
<400> 81
Pro Ser Ile Gly Ile Cys Phe Phe Cys Ser Tyr Ile Ile Ile Ser Phe
1 5 10 15
Leu Ile Val Val Asn Met Tyr Ile Ala Ile Ile Leu Glu Asn Phe
20 25 30
<210> 82
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.5_D4_S6（图 20）
<400> 82
Pro Ala Val Gly Ile Leu Phe Phe Thr Thr Tyr Ile Ile Ile Ser Phe
1 5 10 15
Leu Ile Val Val Asn Met Tyr Ile Ala Ile Ile Leu Glu Asn Phe
20 25 30
<210> 83
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.6_D4_S6（图 20）
<400> 83
Pro Ser Val Gly Ile Phe Phe Phe Val Ser Tyr Ile Ile Ile Ser Phe
1 5 10 15
Leu Ile Val Val Asn Met Tyr Ile Ala Ile Ile Leu Glu Asn Phe
20 25 30
<210> 84
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.7_D4_S6（图 20）
<400> 84
Pro Ser Val Gly Ile Phe Tyr Phe Val Ser Tyr Ile Ile Ile Ser Phe
1 5 10 15
Leu Val Val Val Asn Met Tyr Ile Ala Val Ile Leu Glu Asn Phe
20 25 30
<210> 85
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.8_D4_S6（图 20）
<400> 85
Pro Ala Val Gly Ile Ile Phe Phe Thr Thr Tyr Ile Ile Ile Ser Phe
1 5 10 15
Leu Ile Met Val Asn Met Tyr Ile Ala Val Ile Leu Glu Asn Phe
20 25 30
<210> 86
<211> 31
<212> PRT
<213> 未知
<220>
<223> NaV1.9_D4_S6（图 20）
<400> 86
Pro Gly Ile Ala Thr Ser Tyr Phe Val Ser Tyr Ile Ile Ile Ser Phe
1 5 10 15
Leu Ile Val Val Asn Met Tyr Ile Ala Val Ile Leu Glu Asn Phe
20 25 30
<210> 87
<211> 16
<212> PRT
<213> 未知
<220>
<223> S4 栅极电荷
<400> 87
Pro Thr Leu Gln Thr Ala Arg Thr Leu Arg Ile Leu Lys Ile Ile Pro
1 5 10 15
<210> 88
<211> 16
<212> PRT
<213> 未知
<220>
<223> S4 栅极电荷
<400> 88
Ala Leu Leu Arg Leu Phe Arg Met Leu Arg Ile Phe Lys Leu Gly Lys
1 5 10 15
<210> 89
<211> 17
<212> PRT
<213> 未知
<220>
<223> S4 栅极电荷
<400> 89
Lys Pro Leu Ile Ser Met Lys Phe Leu Arg Pro Leu Arg Val Leu Ser
1 5 10 15
Gln
<210> 90
<211> 22
<212> PRT
<213> 未知
<220>
<223> S4 栅极电荷
<400> 90
Gln Leu Ile Leu Leu Ser Arg Ile Ile His Met Leu Arg Leu Gly Lys
1 5 10 15
Gly Pro Lys Val Phe His
20

Claims (76)

1.一种鉴定人NaX离子通道蛋白(NaX)调节剂的方法，所述方法包括：
a)提供突变型人NaX，其中人NaX的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换；
b)在潜在NaX调节剂的存在下对所述突变型人NaX进行离子通道测定；以及
c)如果在所述潜在调节剂的存在下所述测定中的所述突变型人NaX的活性高于或低于在所述潜在调节剂不存在下的活性，则将所述潜在调节剂鉴定为人NaX调节剂。
2.根据权利要求1所述的方法，其中所述突变型人NaX包含来自人或哺乳动物NaV的DIIIS1-S6区域的全部或部分，任选地其中经置换的人NaX的DIII结构域的所述至少一部分包含或由以下组成：(a)DIII(残基920-1200)，(b)DIII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)，(c)DIII VSD3、S4-S5接头和S5(残基920-1078)，(d)DIII和DII-DIII接头(残基733-1200)，(e)DIII和DIII-DIV接头(920-1237)，(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。
3.根据权利要求1或2所述的方法，其中经所述人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的所述至少-部分不包含S5且/或不包含S6。
4.根据权利要求1至3中任一项所述的方法，其中经所述人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的所述至少一部分不包含S1、S2、S3、S4和/或S4-S5接头中的任一者。
5.根据权利要求1至4中任一项所述的方法，其中人NaX的DIII结构域的所述至少一部分经哺乳动物Nav1.1、哺乳动物Nav1.2、哺乳动物Nav1.3、哺乳动物Nav1.4、哺乳动物Nav1.5、哺乳动物Nav1.6、哺乳动物Nav1.7、哺乳动物Nav1.8、哺乳动物Nav1.9、人Nav1.1、人Nav1.2、人Nav1.3、人Nav1.4、人Nav1.5、人Nav1.6、人Nav1.7、人Nav1.8或人Nav1.9的对应部分置换。
6.根据权利要求5所述的方法，其中人NaX的DIII结构域的所述至少一部分经人Nav1.7的对应部分置换。
7.根据权利要求6所述的方法，其中所述突变型人NaX是嵌合构建体2、嵌合构建体3、嵌合构建体7、嵌合构建体9、嵌合构建体11、嵌合构建体15或嵌合构建体19，或者其中所述突变型人NaX包含SEQ ID No：3、4、8、10、12、16或20中的一者的氨基酸序列。
8.一种鉴定人NaX离子通道蛋白(NaX)调节剂的方法，所述方法包括：
a)提供突变型人NaX，其包含以甘氨酸、脯氨酸或者极性或带电残基对人NaX的两个、三个或四个S6 α螺旋中的每个S6 α螺旋上的至少一个残基的取代；
b)在潜在调节剂的存在下对所述突变型人NaX进行离子通道测定；以及
c)如果在所述潜在调节剂的存在下所述测定中的所述突变型人NaX的活性高于或低于在所述潜在调节剂不存在下的活性，则将所述潜在调节剂鉴定为人NaX调节剂。
9.根据权利要求8所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对所述四个S6 α螺旋中的两个S6 α螺旋上的至少一个残基的取代。
10.根据权利要求8所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对所述四个S6 α螺旋中的三个S6 α螺旋上的至少一个残基的取代。
11.根据权利要求8所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对全部四个S6 α螺旋上的至少一个残基的取代。
12.根据权利要求8所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对所述四个S6 α螺旋中的至少三个S6 α螺旋上的氨基酸残基的一个取代。
13.根据权利要求8所述的方法，其中所述突变型人NaX包含在SEQ ID NO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。
14.根据权利要求8所述的方法，其中所述突变型人NaX包含在SEQ ID NO：1的以下区段中的至少三个区段内以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。
15.根据权利要求8所述的方法，其中所述突变型人NaX包含在SEQ ID NO：1的以下区段中的每个区段内以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。
16.根据权利要求8所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对在残基L390、F724、I1189和I1492中的两者或更多者处的氨基酸残基的取代。
17.根据权利要求8所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对在残基L390、F724、I1189和I1492中的三者或更多者处的氨基酸残基的取代。
18.根据权利要求17所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基L390、F724和I1189或对氨基酸残基F724、I1189和I1492的取代。
19.根据权利要求18所述的方法，其中与野生型人NaX相比，所述突变型人NaX包含取代F724Q、I1189T和I1492T。
20.根据权利要求18所述的方法，其中与野生型人NaX相比，所述人NaX包含取代L390E、I1189E和I1492E。
21.根据权利要求1至20中任一项所述的方法，其中所述离子通道测定是在所述突变型人NaX的经鉴定的调节剂的存在下进行。
22.根据权利要求21所述的方法，其中所述突变型人NaX的所述经鉴定的调节剂是河豚毒素(TTX)、奎尼丁、氟卡尼、丁卡因或利多卡因中的一者或多者。
23.根据权利要求1至22中任一项所述的方法，其中所述潜在调节剂是肽或大环或抗体。
24.根据权利要求1至22中任一项所述的方法，其中所述潜在调节剂是小分子。
25.根据权利要求1至24中任一项所述的方法，其进一步包括确定在部分(c)中鉴定的所述潜在调节剂与所述突变型人NaX和野生型人NaX中的任一者或两者的结合亲和力。
26.根据权利要求25所述的方法，其中所述潜在调节剂以10μM或更低、10μM至50nM、10μM至500nM、1μM或更低、1μM至50nM或者100nM或更低的EC50或IC50与突变型人NaX和野生型人NaX中的任一者或两者结合。
27.根据权利要求1至26中任一项所述的方法，其中所述离子通道测定是膜片钳或自动化膜片钳测定、离子通量测定或者离子敏感或电压敏感染料测定。
28.根据权利要求1至27中任一项所述的方法，其中在部分(c)中鉴定的所述潜在调节剂使所述测定中的所述突变型人NaX的活性降低至少25％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％或至少90％。
29.根据权利要求1至28中任一项所述的方法，其中在部分(c)中鉴定的所述潜在调节剂以10nM至500μM、50nM至500μM、10nM至50μM、100nM至500μM、100nM至50μM、1至500μM、1至50μM、10至500μM或50至250μM的半最大浓度降低所述测定中的所述突变型人NaX的活性。
30.根据权利要求1至27中任一项所述的方法，其中在部分(c)中鉴定的所述潜在调节剂增加所述测定中的所述突变型人NaX的活性。
31.一种通过根据权利要求1至30中任一项所述的方法鉴定的人NaX的调节剂，所述调节剂任选地是肽、大环、小分子或抗体。
32.一种突变型人NaX离子通道(NaX)蛋白，其中人NaX的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换。
33.根据权利要求32所述的突变型人NaX，其中所述突变型人NaX包含来自人或哺乳动物NaV的DIII S1-S6区域的全部或部分，任选地其中经置换的人NaX的DIII结构域的所述至少一部分包含或由以下组成：(a)DIII(残基920-1200)，(b)DIII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)，(c)DIII VSD3、S4-S5接头和S5(残基920-1078)，(d)DIII和DII-DIII接头(残基733-1200)，(e)DIII和DIII-DIV接头(920-1137)，(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。
34.根据权利要求32或33所述的突变型人NaX，其中经所述人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的所述至少一部分不包含S5且/或不包含S6。
35.根据权利要求32至34中任一项所述的突变型人NaX，其中经所述人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的所述至少一部分不包含S1、S2、S3、S4和/或S4-S5接头中的任一者。
36.根据权利要求32至35中任一项所述的突变型人NaX，其中人NaX的DIII结构域的所述至少一部分经哺乳动物Nav1.1、哺乳动物Nav1.2、哺乳动物Nav1.3、哺乳动物Nav1.4、哺乳动物Nav1.5、哺乳动物Nav1.6、哺乳动物Nav1.7、哺乳动物Nav1.8、哺乳动物Nav1.9、人Nav1.1、人Nav1.2、人Nav1.3、人Nav1.4、人Nav1.5、人Nav1.6、人Nav1.7、人Nav1.8或人Nav1.9的对应部分置换。
37.根据权利要求36所述的突变型人NaX，其中人NaX的DIII结构域的所述至少一部分经人Nav1.7的对应部分置换。
38.根据权利要求37所述的突变型人NaX，其中所述突变型人NaX是嵌合构建体2、嵌合构建体3、嵌合构建体7、嵌合构建体9、嵌合构建体11、嵌合构建体15或嵌合构建体19，或者其中所述突变型人NaX包含SEQ ID No：3、4、8、10、12、16或20中的一者的氨基酸序列。
39.一种突变型人NaX离子通道(NaX)蛋白，其包含以甘氨酸、脯氨酸或者极性或带电残基对两个、三个或四个S6 α螺旋中的每个S6 α螺旋上的至少一个残基的取代。
40.根据权利要求39所述的突变型人NaX，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对所述四个S6 α螺旋中的两个S6 α螺旋上的至少一个残基的取代。
41.根据权利要求39所述的突变型人NaX，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对所述四个S6 α螺旋中的三个S6 α螺旋上的至少一个残基的取代。
42.根据权利要求39所述的突变型人NaX，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对全部四个S6 α螺旋上的至少一个残基的取代。
43.根据权利要求39所述的突变型人NaX，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对所述四个S6 α螺旋中的至少三个S6 α螺旋上的氨基酸残基的一个取代。
44.根据权利要求39所述的突变型人NaX，其中所述突变型人NaX包含在SEQ ID NO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。
45.根据权利要求39所述的突变型人NaX，其中所述突变型人NaX包含在SEQ ID NO：1的以下区段中的至少三个区段内以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。
46.根据权利要求39所述的突变型人NaX，其中所述突变型人NaX包含在SEQ ID NO：1的以下区段中的每个区段内以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。
47.根据权利要求39所述的突变型人NaX，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对在残基L390、F724、I1189和I1492中的两者或更多者处的氨基酸残基的取代。
48.根据权利要求39所述的突变型人NaX，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对在残基L390、F724、I1189和I1492中的三者或更多者处的氨基酸残基的取代。
49.根据权利要求48所述的突变型人NaX，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基L390、F724和I1189或对氨基酸残基F724、I1189和I1492的取代。
50.根据权利要求49所述的突变型人NaX，其中与野生型人NaX相比，所述突变型人NaX包含取代F724Q、I1189T和I1492T。
51.根据权利要求49所述的突变型人NaX，其中与野生型人NaX相比，所述人NaX包含取代L390E、I1189E和I1492E。
52.一种确定调节人离子通道蛋白的活性的测试分子是否调节人NaX离子通道(NaX)蛋白的活性的方法，所述方法包括：
a)提供突变型人NaX，其中人NaX的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换；
b)在所述测试分子的存在下对所述突变型人NaX进行离子通道测定；以及
c)如果在所述测试分子的存在下所述测定中的所述突变型人NaX的活性高于或低于在所述测试分子不存在下的活性，则确定所述测试分子为人NaX调节剂；以及任选地，
d)如果根据部分(c)所述测试分子不是人NaX调节剂，则选择所述测试分子进行额外筛选。
53.一种确定调节人离子通道蛋白的活性的测试分子是否调节人NaX离子通道(NaX)蛋白的活性的方法，所述方法包括：
a)确定所述测试分子调节所述人离子通道蛋白的活性；
b)提供突变型人NaX，其中人NaX的DIII结构域的至少一部分经人或哺乳动物NaV蛋白(NaV)的DIII结构域的对应部分置换；
c)在所述测试分子的存在下对所述突变型人NaX进行离子通道测定；以及
d)如果在所述测试分子的存在下所述测定中的所述突变型人NaX的活性高于或低于在所述测试分子不存在下的活性，则确定所述测试分子为人NaX调节剂；以及任选地，
e)如果根据部分(d)所述测试分子不是人NaX调节剂，则选择所述测试分子进行额外筛选。
54.根据权利要求52或53所述的方法，其中所述突变型人NaX包含来自人或哺乳动物NaV的DIII S1-S6区域的全部或部分，任选地其中经置换的人NaX的DIII结构域的所述至少一部分包含或由以下组成：(a)DIII(残基920-1200)，(b)DIII电压传感器结构域III(VSD3)和S4-S5接头(残基920-1058)，(c)DIII VSD3、S4-S5接头和S5(残基920-1078)，(d)DIII和DII-DIII接头(残基733-1200)，(e)DIII和DIII-DIV接头(920-1237)，(f)DIII和DII-DIII接头和DIII-DIV接头(733-1237)。
55.根据权利要求52、53或54所述的方法，其中经所述人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的所述至少一部分不包含S5且/或不包含S6。
56.根据权利要求52至55中任一项所述的方法，其中经所述人或哺乳动物NaV的DIII结构域的对应部分置换的人NaX的DIII结构域的所述至少一部分不包含S1、S2、S3、S4和/或S4-S5接头中的任一者。
57.根据权利要求52至56中任一项所述的方法，其中人NaX的DIII结构域的所述至少一部分经哺乳动物Nav1.1、哺乳动物Nav1.2、哺乳动物Nav1.3、哺乳动物Nav1.4、哺乳动物Nav1.5、哺乳动物Nav1.6、哺乳动物Nav1.7、哺乳动物Nav1.8、哺乳动物Nav1.9、人Nav1.1、人Nav1.2、人Nav1.3、人Nav1.4、人Nav1.5、人Nav1.6、人Nav1.7、人Nav1.8或人Nav1.9的对应部分置换。
58.根据权利要求57所述的方法，其中人NaX的DIII结构域的所述至少一部分经人Nav1.7的对应部分置换。
59.根据权利要求58所述的方法，其中所述突变型人NaX是嵌合构建体2、嵌合构建体3、嵌合构建体7、嵌合构建体9、嵌合构建体11、嵌合构建体15或嵌合构建体19，或者其中所述突变型人NaX包含SEQ ID No：3、4、8、10、12、16或20中的一者的氨基酸序列。
60.一种确定调节人离子通道蛋白的活性的测试分子是否调节人NaX离子通道(NaX)蛋白的活性的方法，所述方法包括：
a)提供突变型人NaX，其包含以甘氨酸、脯氨酸或者极性或带电残基对人NaX的两个、三个或四个S6 α螺旋中的每个S6 α螺旋上的至少一个残基的取代；
b)在所述测试分子的存在下对所述突变型人NaX进行离子通道测定；以及
c)如果在所述测试分子的存在下所述测定中的所述突变型人NaX的活性高于或低于在所述测试分子不存在下的活性，则确定所述测试分子为人NaX调节剂；以及任选地，
d)如果根据部分(c)所述测试分子不是人NaX调节剂，则选择所述测试分子进行额外筛选。
61.一种确定调节人离子通道蛋白的活性的测试分子是否也调节人NaX离子通道(NaX)蛋白的活性的方法，所述方法包括：
a)确定所述测试分子调节所述人离子通道蛋白的活性；
b)提供突变型人NaX，其包含以甘氨酸、脯氨酸或者极性或带电残基对人NaX的两个、三个或四个S6 α螺旋中的每个S6 α螺旋上的至少一个残基的取代；
c)在所述测试分子的存在下对所述突变型人NaX进行离子通道测定；以及
d)如果在所述测试分子的存在下所述测定中的所述突变型人NaX的活性高于或低于在所述测试分子不存在下的活性，则确定所述测试分子为人NaX调节剂；以及任选地，
e)如果根据部分(d)所述测试分子不是人NaX调节剂，则选择所述测试分子进行额外筛选。
62.根据权利要求60或61所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对所述四个S6 α螺旋中的两个S6 α螺旋上的至少一个残基的取代。
63.根据权利要求60或61所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对所述四个S6 α螺旋中的三个S6 α螺旋上的至少一个残基的取代。
64.根据权利要求60或61所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对全部四个S6 α螺旋上的至少一个残基的取代。
65.根据权利要求60或61所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对所述四个S6 α螺旋中的至少三个S6 α螺旋上的氨基酸残基的一个取代。
66.根据权利要求60或61所述的方法，其中所述突变型人NaX包含在SEQ ID NO：1的以下区段中的至少两个区段内以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。
67.根据权利要求60或61所述的方法，其中所述突变型人NaX包含在SEQ ID NO：1的以下区段中的至少三个区段内以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。
68.根据权利要求60或61所述的方法，其中所述突变型人NaX包含在SEQ ID NO：1的以下区段中的每个区段内以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基的一个或两个取代：残基383-397(在结构域I(D1)的S6中)、残基717-731(在DII的S6中)、残基1182-1196(在DIII的S6中)和/或残基1485-1499(在DIV的S6中)。
69.根据权利要求60或61所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对在残基L390、F724、I1189和I1492中的两者或更多者处的氨基酸残基的取代。
70.根据权利要求60或61所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对在残基L390、F724、I1189和I1492中的三者或更多者处的氨基酸残基的取代。
71.根据权利要求70所述的方法，其中所述突变型人NaX包含以甘氨酸、脯氨酸或者极性或带电残基对氨基酸残基L390、F724和I1189或对氨基酸残基F724、I1189和I1492的取代。
72.根据权利要求71所述的方法，其中与野生型人NaX相比，所述突变型人NaX包含取代F724Q、I1189T和I1492T。
73.根据权利要求71所述的方法，其中与野生型人NaX相比，所述人NaX包含取代L390E、I1189E和I1492E。
74.一种通过根据权利要求52至73中任一项所述的方法鉴定的分子，所述分子任选地是肽、大环、小分子或抗体。
75.一种分子复合物，其包含根据权利要求32至51中任一项所述的嵌合型或突变型人NaX和根据权利要求31所述的调节剂或根据权利要求52、53、60或61所述的测试分子。
76.一种用于测定人NaX的活性或用于鉴定与人NaX结合的人NaX调节剂和/或分子的试剂盒，所述试剂盒包括根据权利要求32至51中任一项所述的突变型人NaX，并且进一步包括以下中的至少一者：一种或多种用于执行离子通道测定的试剂、所述突变型人NaX的调节剂以及使用说明。
CN202280038599.0A 2021-05-30 2022-05-27 突变型人nax蛋白和筛选方法 Pending CN117616038A (zh)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US202163195039P 2021-05-30 2021-05-30
US63/195,039 2021-05-30
PCT/US2022/031302 WO2022256251A1 (en) 2021-05-30 2022-05-27 Mutant human nax proteins and screening methods

Publications (1)

Publication Number Publication Date
CN117616038A true CN117616038A (zh) 2024-02-27

Family

ID=82115716

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202280038599.0A Pending CN117616038A (zh) 2021-05-30 2022-05-27 突变型人nax蛋白和筛选方法

Country Status (5)

Country Link
EP (1) EP4347633A1 (zh)
JP (1) JP2024520478A (zh)
CN (1) CN117616038A (zh)
TW (1) TW202306973A (zh)
WO (1) WO2022256251A1 (zh)

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7094600B2 (en) * 2003-06-26 2006-08-22 The Research Foundation Of State University Of New York Screen for sodium channel modulators
CA2745371C (en) * 2008-12-22 2016-01-05 Universite Laval Methods, reagents and kits for the identification of modulators of voltage-gated sodium channels
WO2013016418A2 (en) * 2011-07-27 2013-01-31 The Regents Of The University Of Michigan Ion channel probes and methods of use thereof

Also Published As

Publication number Publication date
EP4347633A1 (en) 2024-04-10
JP2024520478A (ja) 2024-05-24
WO2022256251A1 (en) 2022-12-08
TW202306973A (zh) 2023-02-16

Similar Documents

Publication Publication Date Title
Noreng et al. Structure of the human epithelial sodium channel by cryo-electron microscopy
Confalonieri et al. Tyrosine phosphorylation of Eps15 is required for ligand-regulated, but not constitutive, endocytosis
Hou et al. Cryo-EM structure of the calcium release-activated calcium channel Orai in an open conformation
Bodrug et al. The kinesin-5 tail domain directly modulates the mechanochemical cycle of the motor domain for anti-parallel microtubule sliding
Ruan et al. Structures of the TRPM5 channel elucidate mechanisms of activation and inhibition
Noland et al. Structure-guided unlocking of NaX reveals a non-selective tetrodotoxin-sensitive cation channel
Araya-Secchi et al. The prolactin receptor scaffolds Janus kinase 2 via co-structure formation with phosphoinositide-4, 5-bisphosphate
CN117616038A (zh) 突变型人nax蛋白和筛选方法
US20050191612A1 (en) Method of screening insulin content enhancer
Ye et al. Structural heterogeneity of the ion and lipid channel TMEM16F
Strauss et al. Structural basis of allosteric modulation of metabotropic glutamate receptor activation and desensitization
Kienker et al. Topography of the TH5 segment in the diphtheria toxin T-domain channel
Hussain et al. Structural insights into the organization and channel properties of human Pannexin isoforms 1 and 3
Duan et al. Cryo-EM structure of the receptor-activated TRPC5 ion channel at 2.9 angstrom resolution
Maity et al. Cryo-EM structure of OSCA1. 2 from Oryza sativa: Mechanical basis of hyperosmolality-gating in plants
Shi et al. Human P2X4 receptor gating is modulated by a stable cytoplasmic cap and a unique allosteric pocket
CN109791156B (zh) 鉴别表位的方法
Körbel et al. Phosphoprotein profiling of erythropoietin receptor‐dependent pathways using different proteomic strategies
Huang et al. Inhibition and transport mechanisms of the ABC transporter hMRP5
WO2011068801A2 (en) Methods for identifying compounds that modulate ion channel activity of a kir channel
Azumaya Structure-function analyses of ion channels involved in glutamatergic signaling
CN117642632A (zh) Nalcn通道蛋白及其调节剂
Tonggu Structural studies of BK ion channel in a lipid environment
Suo Structural and Functional Studies on Noxious Stimuli Sensing of the Transient Receptor Potential Ankyrin 1 Channel
Mortazavi Structural Studies of the KCa3. 1-CaM Channel Complex, a Critical Component of T-Cell Function

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
REG Reference to a national code

Ref country code: HK

Ref legal event code: DE

Ref document number: 40105807

Country of ref document: HK