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

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.