蝰蛇(缅甸亚种)毒促凝—纤溶双相活性组分FⅥbb的分离纯化及生物活性  

Purification,biochemical properties,and activities of FⅥbb, a procoagulant-fibrinolytic biphasic active component from daboia russelli siamensis (myanmar) venom

作　　者：王睿[1] 朱玮玮[1] 林熙[1] 孙燕[2] 

机构地区：[1]暨南大学医学院药理学系,广东广州510632 [2]中山大学中山医学院药理学教研室,广东广州510080

出　　处：《中国药理学通报》2013年第6期802-808,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81000209);教育部科学技术研究重点资助项目(No 210255);暨南大学科研培育与创新基金资助项目(No 21609304)

摘　　要：目的从蝰蛇(缅甸亚种)毒分离纯化一种促凝—纤溶双相活性组分FⅥbb,并研究其理化性质和生物活性。方法应用CM-Sephadex C-50阳离子交换层析Sephadex G-150(超细)凝胶过滤层析Chelating Sepharose Fast Flow金属离子螯合亲和层析分离纯化蛇毒,反相HPLC检测组分FⅥbb纯度,采用MALDI质谱测定法测定分子量,以发色底物法、纤维平板法和SDS-PAGE测定FⅥbb的酶学特征和生物活性。结果从蝰蛇(缅甸亚种)毒分离得到的促凝—纤溶双相活性组分FⅥbb为单体蛋白,相对分子质量为59 138,最适温度为40℃,最适pH为10.0,为金属蛋白酶。结论应用CM-Sephadex C-50阳离子交换层析和Chelating Sepha-rose Fast Flow金属离子螯合亲和层析等方法可以从蝰蛇(缅甸亚种)毒纯化出促凝—纤溶双相活性组分FⅥbb,具有促凝—纤溶双相活性。Aim To purify F Ⅵbb, a procoagulant-fi-brinolytic biphasic active component from daboia rus-selli siamensis （myanmar） venom ,and illustrate its bi- ochemical and biological characters. Methods FⅥbb was purified from daboia russelli siamensis （myanmar） venom by a combination of cation exchange chromatog- raphy on CM-Sephadex C-50, gel filtration on Sepha- dex G-150 （ superfine ） , Znion affinity chromatography on Chelating Sepharose Fast Flow. The purity of FⅥbb was determined by reversed-phase HPLC. The molecu- lar weight of F Ⅵ bb was determined by MALDI mass spectrometry. The procoagulant-fibrinolytic biphasic active and the enzymological characteristics of F Ⅵ bb were determined based on chromogenic substrates,SDS-PAGE and fibrin plate method. Results FⅥbb was a metalloprotease, which was achieved by chroma- tography with a molecular weight of 59 138 and an iso- electric point of 4.6. The suitable temperature and pH range of FⅥbb was 40 ℃ and 10. Conclusion F Ⅵ bb can be successfully isolated and purified by CM- Sephadex C-50 ion exchange chromatograph and Chela- ting Sepharose Fast Flow affinity chromatograph. It has procoagulant-fibrinolytic biphasic activity.

关 键 词：蝰蛇 FⅥbb促凝活性 纤溶活性 金属蛋白酶 蛋白 分离 

分 类 号：R345.9[医药卫生—基础医学] R973.1