摘要
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.
作者
肖昌华
何丽芬
唐绍宗
杨筠茹
董馥明
Xiao Changhua;He Lifeng;Tan Shaozong;Yang Junru;Dong Fuming(Kunming Institute of Zoology,Acadmia Sinica)
关键词
尖吻蝮蛇
凝血酶样
电泳
凝结活性
精氨酸酯酶
Agkisirodon acutus
activity of coagulation
arginineesterase
electrophoresis.