白唇竹叶青蛇(Trimeresurus albolabris)毒降纤酶的分离纯化以及性质  

Purification and characterization of fibrinolytic enzyme from the snake (Trimeresurus albolabris) venom

作　　者：邓敏[1] 余晓东[1] 陈夏[1] 李卉[1] 林亦心[1] 和七一[1] 柳建平[1] 

机构地区：[1]重庆师范大学生命科学学院重庆市生物活性物质工程研究中心重庆市动物生物学重点实验室,重庆400047

出　　处：《中国生化药物杂志》2008年第3期156-160,共5页Chinese Journal of Biochemical Pharmaceutics

基　　金：重庆市自然科学基金重点项目(CSTC2006BA5032)

摘　　要：目的从白唇竹叶青蛇(T.albolabris)毒中分离纯化无出血作用的降纤活性组分,探讨其理化性质及部分生物功能。方法用DEAE-SephadexA-25,SephadexG-100和CM-SephadexC-50三步色谱法进行分离纯化。SDS-PAGE和HPLC鉴定其纯度和相对分子质量,平板法测定其降纤活性。结果从白唇竹叶青蛇毒中分离纯化获得单一的降纤组分,能迅速水解纤维蛋白原或纤维蛋白原Aα链,缓慢水解Bβ链,而对γ链无作用,SDS-PAGE鉴定其相对分子质量为56000。EDTA能抑制其纤维蛋白原水解活性,而PMSF、β-巯基乙醇对其活性无影响,提示该组分为单链α金属蛋白酶。结论从白唇竹叶青蛇毒中分离纯化得到1种无出血作用且降纤活性强的新蛇毒降纤酶。Purpose To find out the fibrinolytic enzyme component without hemorrhagic activity from snake venom（ Trimeresurus albolabris ） and to study its physical-chemical properties and biological functions. Methods the fibrinolytic enzyme component was isolated and purified by column chromatography of DEAE-SephadexA- 25, SephadexG-100 and CM-SephadexC-50, analyzed and measured by SDS-PAGE and HPLC chromatography, and its activity was assayed by agarose plate method. Results the fibrinolytic enzyme purified from snake venom（ T. albolabris） cleaved Aα and Bβ, but not γ chains of fibrinogen＇or fibrin. Its molecular weight was 56 kDa with SDS-PAGE. Its fibrinolysis activity was inhibited by EDTA, but not PMSF（Phenylmethyl Sulfonyl Fluoride）and β-mercaptoethanol, and these indicated that it may be a metalloproteinase with a single chain. Conclusion A novel fibrinolytic enzyme, with the strong fibrinolysis activity and without hemorrhagic activity, was obtained from the venom of snake（ T. albolabris ）in China.

关 键 词：降纤酶 蛇毒 白唇竹叶青蛇 金属蛋白酶 

分 类 号：R282.74[医药卫生—中药学] R996.3[医药卫生—中医学]