尖吻蝮(Agkistrodon acutus) 蛇毒凝血酶样成分的研究Ⅰ.分离纯化及其理化特性的测定

STUDIES ON THE THROMBIN-LIKE PRINCIPLES OF AGKISTRODON ACUTUS VENOM Ⅰ. ISOLATION AND PURIFICATION OF ITS IDENTIFICATION OF PHYSICAL AND CHEMICAL PROPERTY

尖吻蝮蛇毒经DEAE—sephadex A-50柱层析分离得15个蛋白组分,将其具有凝血酶样活性组分再经DEAE—celluiose DE_(52),QAE—sephadex A-25纯化,再以Sephadex G-25脱盐得精制的凝血酶样活性成份。经聚丙烯酰胺凝胶电泳鉴定为四条迁移率不同的单一蛋白带;再经SDS—聚丙烯酰胺凝胶电泳测得四种凝血酶样酶(TLE)的分子量,TLE;为33500,TLE;为68000,TLE;为19500,TLE;为30000;经等电聚焦电泳测得等电点(PL)分别为4.1,4.8,3.9,3.4,经氨基酸组成分析,四种TLE均由相同的17种氨基酸组成,但其组成氨基酸的数量则有一定的差异。结果表明,从尖蝮蛇毒中分离纯化的这四种蛋白成份,它们都具凝血(或凝结)活性,但在层析,电泳、组成氨基酸的含量等方面则存在显著的差异,因而它们是四种不同的TLE。这是本实验室首先发现并分离得到的,还未见有同时从尖蝮蛇毒中分离、纯化得到四个不同的凝血酶样酶的报道。

Fifteen fractions of venom of Agkistrodon acutus were obtained through DEAE-sephadex A-50 column chromatography.Fractions with thrombin-like activity were purified further on DEAE-cellulose DE52,QAE-sephadex A-25 column chromatography and desalted on sephadex G-25.Single band were obtained and identified by polyacrlamide gel electrophoresis.Their molecular weights were TLE1 MW=33500,TLE2 MW=68000,TLE3 MW=19500,TLE4=30000 by SDS-polyacracrlamide gel electrophoresis.Isoelectric point(P1)were 4.1,4.8,3.9,3.4.(Containing the same 17 kinds of amino-acids but di ffernt in the amount of each kind.)The result shows that four purified TLEs have the activity of coagulation and the activity of arginineestcrase.