尖吻蝮蛇毒无出血活性纤维蛋白溶解酶的纯化及其理化性质研究

目的：从蝮亚科尖吻蝮蛇毒(Agkistrodon acutus venom)中分离纯化出无出血活性纤维蛋白溶解酶(Non—hemonrrhagic fibrinolytic enzyme,NHFLE)并对其理化性质进行研究。方法：应用凝胶过滤和离子交换柱层析法分离纯化尖吻蝮蛇毒NHFLE，用纤维蛋白平板法测定其纤溶活力，用SDS—聚丙烯酰胺凝胶电泳(PAGE)测定其相对分子质量，用皮内出血实验测定其出血活性。结果：从短亚科尖吻蝮蛇毒中分离出一种单一组分的纤维蛋白溶解酶，它的相对分子质量为25000，没有出血活性，在普通纤维蛋白平板和加热平板上都能溶解纤维蛋白，相对活性为粗毒的3．2倍。结论：从尖吻蝮蛇毒中分离出相对分子质量为25000的纯化蛋白是具有直接溶解纤维蛋白活性，而无出血活性的单一的纤维蛋白溶解酶。

PURIFICATION AND CHARACTERIZATION OF NON-HEMORRHAGIC FIBRINOLYTIC ENZYME FROM AGKISTRODON ACUTUS

Objective:To purify non hemorrhagic fibrinolytic enzyme(NHFLE) from Agkistrodon acutus venom and study their physicochemical properties .Methods:We purify non hemorrhagic fibrinolytic enzyme by filtration and anion exchange chromatography and determined fibrinolytic activity by fibrin plate,calculated molecular weight by SDS PAGE;and measured heorrhagic activity on animals.Result:By means of DEAE Sepharose CL 6B ,Sephadex G 75 chromatography and TSK GEL G-2000SW high performance gel filtration,a purified protein with fibrinolytic activity was obtained from the venom of Agkistrodon acutus.The protein was homogeneous as judged by electrophresis.Its molecular weight was estimated to be 25 000 without any hemorrhagic activity. The NHFLE had directly fibrinolytic activity and The specific fibrinolytic activity of the protein was 3.2 time higher than that of crude venom.Conclusion:The NHFLE from Agkistrodon acutus venom has directly fibrinolytic activity. Its molecular weight is estimated to be 25 000 without any hemorrhagic activity.