基因重组蛇毒纤溶因子rFII的酶学特性研究

Studies on Enzyme Characteristics of Recombinant Fibrinolytic Enzyme of Agkistrodon acutus Venom

目的研究重组蛇毒纤溶因子的酶学特性。方法通过SDS-PAGE分析其分子量、纯度,用SDS-PAGE方法检测其对纤维蛋白原的水解能力,用分光光度法检测其水解azocasein的能力和PMSF及EDTA对蛇毒纤溶因子的作用。结果蛇毒重组纤溶因子rFII是一个分子量在28000的蛋白酶。它呈时间和浓度依赖性水解纤维蛋白原的Aα、Bβ、γ亚基,EDTA和PMSF均可以抑制该因子水解azocasein的能力。结论rFII是一种纤维蛋白原酶,它的酶活性可同时被EDTA和PMSF所抑制。

Objective To characterize the recombinant fibrinolytic enzyme (rFII) of Agkistrodon acutus Venom. Methods The molecular weight and fibrinogenolytic activity were determinaed by SDS-PAGE. Azocasein was used as substrate to investigate the caseinolytic activity and the effects of EDTA and PMSF on rFII. Results rFII is a proteinase with a molecular weight of 28000. It acted on the Aα, Bβ,γ chain of fibrinogen directly. The caseinolytic activity was inhibited by EDTA and PMSF. Conclusions rFII is a recombinant proteinase which can hydrolyze fibrinogen. The enzymatic activity can be inhibited by EDTA and PMSF.