雷氏大疣蛛毒素溶栓活性测定及纤溶蛋白的分离

ISOLATION OF FIBRINOLYSIN AND DETERMINATION OF THE SPIDER VENOM FROM MACROTHELE RAVEN

[目的]检测雷氏大疣蛛毒素的体外溶栓活性及分离雷氏大疣蛛毒素中的纤溶酶组分。[方法]应用人血检测雷氏大疣蛛毒素的体外溶栓活性;用反相层析法和阳离子交换法分离雷氏大疣蛛毒素,应用纤维蛋白平板检测毒素及分离组分的纤溶酶活性。[结果]体外溶栓实验显示,不同浓度的雷氏大疣蛛毒素水溶液均可溶解人血凝块,浓度越高溶解作用越强。经纤维蛋白平板法检测,雷氏大疣蛛毒素具有良好的纤溶作用。反相层析法分离雷氏大疣蛛毒素收集得14种组分,其中4种具有纤溶活性。取其中分离效果较好的组分2和10利用阳离子交换法再分离,各收集了2种组分,其中2-2和10-2具纤溶活性且SDS-PAGE显示一条带,达到了电泳纯。[结论]雷氏大疣蛛毒素存在具纤溶酶活性的蛋白(多肽)组分,应用反相层析和阳离子交换的反法能较好的分离雷氏大疣蛛毒素组分。

[Objective] To determine the activity of vitro thrombolysis of spider venom from Macrothele raveni, and to isolate the composition of fibrinolysin of spider venom from Macrothele raveni. [Methods] The spider venom from Maerothele raveni was isolated by reversion phase chromatography and cation exchange, and the toxin and plasmin activity were detected by fibrin plate method. [ Results] The vitro thrombolysis test demonstrated that the water solution with different concentration of spider venom from Macrothele raven could solve the human blood clot, and the solution traction was increased with the concentration. The spider venom from Macrothele raveni had favorable thrombolyie activity. Four of the 14 fractions isolated from Maerothele raven collected by RP-chromatography had the thrombolyic activity. Fraction 2 and 10 isolated by RP-chromatography were isolated by cation-exchange chromatography again. 2-2 and 10-2 showed thrombolyie activity by means of fibrin plate, and both of them showed one zone which had reach the purification of electrophoresis detected by SDS-PAGE. [ Conclusion] Spider venom from Macrothele raveni had the protein fraction ofplasmin activity, and the components of the spider venom can be isolated efficiently by the method of RP-ehromatography and cation-exchange chromatography.