中国药理学通报2011，Vol.27Issue(6)：818-823,6.DOI:10.3969/j.issn.1001-1978.2011.06.019
原矛头蝮蛇毒中一个新颖双链纤溶酶的分离纯化与性质研究
Purification and characterization of a novel double-stranded serine protease from Protobothrops mucrosquamatus venom
摘要
Abstract
Aim To discover the new or novel fibrinolysin from Protobothrop.s mucrosquamatus venom. The physico-chemical property and biological activity of the interest enzyme were investigated for better understanding of its role in snakebite and evaluating its clinical potential. Methods The object protein was isolated by chromotography techniques. Assays were performed to measure molecular mass. isoelectric point, peptide mass fingerprinting analysis, variety of proteolytic activity, anticomplementary activity, edema-inducing activity, hemorrhagic activity, and mice tail bleeding time. Results By anion exchange chromatography,gel filtration chromatography, and affinity chromatography, an acidic fibrinolysin PMSP-A was purified from Protobothrops mucro.squamatus venom. It consists of two non-equal polypeptides connected by covalent bond. The isoelectric points of the two subunits were 5.7 and 6. 1, respectively. The molecular weight of PMSP-A was determined to be 26. 1 ku and 25. 3 ku by SDS-PAGE and gel filtration, respectively. Peptide mass fingerprinting analysis revealed that PMSP-A shared sequence similarity with the blood clotting factor from Trimeresurus flavoviridi.s. PMSP-A degraded Bβ- chain and Aα-chain of fibrinogen in order. This fibrinogenolytic activity was inhibited by PMSF and 1, 10phenanthroline. but not by EDTA, EGTA, and SBTI.PMSP-A hydrolyzed fibrin and arginine ethyl ester, but no hydrolytic activity against azocasein. PMSP-A induced slight edema of mice paw. No hemorrhagic activity was observed. Interestingly, PMSP-A prolonged the bleeding time of mice tail vein significantly. Conclusion PMSP-A is a novel double chains serine protease from Protobothrops mucrosquamatus. and it possesses diversified biological activity.关键词
蛇毒丝氨酸蛋白酶/纤溶酶/抗凝活性/纤维蛋白(原)水解活性/抗补体活性/蛇伤/蛇毒/原矛头蝮蛇/烙铁头蛇分类
医药卫生引用本文复制引用
季仁升,孙黔云,乙引..原矛头蝮蛇毒中一个新颖双链纤溶酶的分离纯化与性质研究[J].中国药理学通报,2011,27(6):818-823,6.基金项目
国家自然科学基金资助项目(No 30560035) （No 30560035）
贵州省科技计划资助项目(No黔科合SY(2010)3050号) （No黔科合SY(2010）
贵州省优秀青年科技人才资助项目(No黔科合人字2005-0510) （No黔科合人字2005-0510）
