基因重组蛇毒纤溶因子rFII理化特性及纤溶能力的研究

Biochemical Characterization and Fibrinolytic Activity of rFII, a Recombinant Fibrinolytic Enzyme from Agkistrodon acutus Venom

【目的】研究重组蛇毒纤溶因子的理化特性和纤溶能力。【方法】SDS鄄PAGE及高效液相法分析重组蛇毒纤溶因子rFII相对分子质量、纯度,等电聚焦分析其等电点;纤维平板和SDS鄄PAGE检测重组蛇毒纤溶因子对纤维蛋白及纤维蛋白原的水解能力。【结果】重组蛇毒纤溶因子rFII是一个相对分子质量在28000,等电点9.0的碱性蛋白酶。它水解纤维蛋白原的Aα、Bβ、酌亚基,水解能力随时间和浓度的增加而增加。它对纤维蛋白的水解能力比相同浓度下的纤溶酶强4倍。【结论】rFII是一种高效的水解纤维蛋白原和纤维蛋白的碱性蛋白酶。

Objective]To observe the biochemical characterization and fibrinolytic activity of rFII. [Method]We calculated the molecular weight by SDS-PAGE, determined the pI of rFII by isoelectric focusing and the purity by HPLC. We determined fibrinogenolytic activity by SDS-PAGE and fibrinolytic activity by fibrin plate.[Result]rFII was an alkaline proteinase which had a relative molecular mass of 28 000. It acted on the Aα, Bβ, γ chain of fibrinogen directly. The fibrinolytic activity was 4 times higher than that of plasmin in the same concentration.[Conclusion]rFII was an alkaline proteinase which hydrolyzed fibrinogen and fibrin efficiently.