常用蛋白酶切割位点.doc

常用融合蛋白切割位点PROTEASE COMPANY Protease Capture Thrombin Leu-Val-Pro-ArgGly-SerL-V-P-RG-SAmersham-Biosciences, Novagen,SIGMA, RocheBenzamidine-AgaroseFactor Xa Ile-Glu/Asp-Gly-ArgI-E/D-G-RAmersham-Biosciences, New England Biolabs,RocheBenzamidine-AgaroseEnterokinase Asp-Asp-Asp-Asp-LysD-D-D-D-KNew England Biolabs,Novagen, RocheTrypsin Inhibitor-AgaroseTEV protease Glu-Asn-Leu-Tyr-Phe-GlnGlyE-N-L-YF-QGInvitrogen Life TechnologiesNi-NTA (6His recomb. TEV)PreScission Leu-Glu-Val-Leu-Phe-GlnGly-ProL-E-V-L-F-QG-PAmersham-BiosciencesGSTrap for GST fusion enzymeTAGZyme His-tag removal by Exoproteolytic DigestionQiagenNi-NTA (6His recomb. enzyme)Intein Site dithiothreitol cleavageNew England BiolabsDTT elimination by dialysisHRV 3C Protease Leu-Glu-Val-Leu-Phe-GlnGly-ProL-E-V-L-F-QG-PNovagen Ni-NTA (6His recomb. enzyme)SUMO Protease recognize the tertiary structure of the ubiquitin-like (UBL) protein, SUMO Invitrogen Life Technologies, LifeSensorsNi-NTA (6His recomb. enzyme)Kex-2-Arg-X-Lys/Arg-ArgInvitrogen Life Technologies,Ni-NTA (6His recomb. enzyme)KEX2对arg的专一性高，要求最重要。Arg前为lys效率最高，不切-Arg-lys，Pro影响KEX2切割羧肽酶羧肽酶B可以切割C端的Lys或Arg；羧肽酶A可以切割C端除了Lys、Arg、Pro的氨基酸，但如果倒数第二个氨基酸为Pro两种羧肽酶均不能作用1.胰蛋白酶属肽链内切酶，能把多肽链中Lys和Arg残基中的羧基侧切断。2.胰凝乳蛋白酶（亦称糜蛋白酶）属肽链内切酶，主要切断多肽链中的芳香族氨基酸（Phe、Trp、Tyr）残基的羧基一侧。3.羧肽酶（分A和B型），一般的题目中没有特别指明的话就是两种类型的功