短尾蝮蛇毒纤溶酶原激活剂的分离纯化及活性测定  

Isolation,characterization and biological activity of a plasminogen activator from Gloydius brevicaudus venom

作　　者：张志强[1] 张进华[2] 余瑞铭[1] 李玉娜[1] 陈建济[1] 许云禄[1] 

机构地区：[1]福建医科大学蛇毒研究所医药生物工程中心 [2]福建医科大学附属协和医院药剂科,福建福州350004

出　　处：《中国药理学通报》2007年第12期1639-1644,共6页Chinese Pharmacological Bulletin

基　　金：福建省自然科学基金计划资助项目(NoC0720002)

摘　　要：目的从短尾蝮蛇毒中分离纯化纤溶酶原激活剂(plasminogen activator of Gloydius brevicaudus venom,GBV-PA),对其理化性质及部分生物活性进行研究。方法应用苯甲脒-琼脂糖凝胶(Benzamidine Sepharose6B)亲和层析及Lichrospher C18反相层析柱进行分离纯化,应用SDS-PAGE测定分子量、聚丙烯酰胺凝胶盘状电泳测定等电点,以发色底物法测定生物活性。结果通过亲和层析、反相层析等方法可从短尾蝮蛇毒中分离纯化出纤溶酶原激活剂至电泳纯以上,其相对分子质量约为32.6×103,等电点约为5.2,能特异激活人纤溶酶原为纤溶酶,其比活性为2.87t-PA IU.mg-1;该酶为丝氨酸蛋白酶,对纤维蛋白无亲和性。结论应用亲和层析和反相层析可从短尾蝮蛇毒获得一种纤溶酶原激活剂;该酶为丝氨酸蛋白酶,对纤维蛋白无亲和性。Aim To isolate and purify a novel plasminogen activator （ PA ） from Gloydius brevicaudus venom （GBV） and study characterization and biological activities of GBV-PA. Methods Affinity chromatography in Benzamidine Sepharose 6B （ AC ） and Lichrospher C-18 4. 6/250 reversed phase chromatography （RPC） were used for isolation and purification; SDS-PAGE was used to detect molecular weight （MW）; Disc polyacrylamide gel eletrophoresis was used to measure the point of isoelectric （pI）; Chromogenic substrate method was used to observe the biological activities. Results A novel GBV-PA which its purification reached the homogeneity level was isolated and purifled from GBV by AC and RPC ; The MW of the novel GBV-PA was 3.26 × 10^4 and the pI was 5.2 ; The novel GBV-PA activated human plasminogen specifically and the special activity was 2. 87 t-PA IU·mg^-1 ; Moreover, our results indicated that this novel GBV-PA was a serine proteinase which had no affinity to fibrin. Conclusion A novel GBV-PA that can be isolated and purificated from GBV by AC and RPC was proved to be a serine protease and has no affinity to fibrin.

关 键 词：短尾蝮蛇毒 纤溶酶原激活剂 亲和层析 反相层析 理化性质 生物活性 

分 类 号：R345[医药卫生—基础医学] R446.1