【摘要】 目的:从沙蚕Nereis virens的体腔液中纯化、鉴定出一种新的纤溶酶,并对其进行分子表征。方法:运用离子交换层析、凝胶过滤层析等常规色谱方法纯化蛋白酶,采用纤维蛋白板检测法鉴定纤溶活性及活性分布曲线,利用2D-电泳方法检测此纤溶酶的表观分子量和等电点,并检测多种蛋白酶抑制剂对其酶活性的影响,以确定此蛋白酶的类型。结果:获取一种新的具有极强纤维蛋白水解活性的单链蛋白酶,其相对分子质量为29000,等电点为4.5,其蛋白水解活性在pH7.8和45℃时表现为最高,并能被二异丙基氟磷酸(DFP)和甲苯磺酚氟(PMSF)完全性抑制,确证其为一种典型的丝氨酸内肽酶。结论:Nereis virens的体腔液中存在一种纤溶活性极强的单链丝氨酸内肽酶,该纤溶酶对预防和治疗血栓性疾病具有一定的药用价值。更多还原

【Abstract】 Objective To detect the partial molecular characteristics of a novel fibrinolytic protease from the coelomic fluid of Nereis virens identified and purified to homogeneity.Methods The fibrinolytic protease was purified with anion and cation exchange chromatography and gel filtration chromatography.The identification of fibrinolytic activity and active distributed curve had been assessed by method of fibrin plate.Its apparent molecular weight and isoelectric point were analysed by two-dimensional gel electrophoresis(2-DE).The effects of several protease inhibitors on the protease activity were examined,and its protease type was identified.Results A novel and single chain fibrinolytic protease was purified effectively.Its apparent molecular weight and isoelectric point were 29 000 and 4.5,respectively.The proteolytic activity peaked at pH 7.8 and 45℃.The protease was completely inhibited by DFP and PMSF,assessing as a serine protease.Conclusion From the coelomic fluid of Nereis virens,a novel and single chain serine protease with more strong fibrinolytic activity is discovered.The fibrinolytic protease has a medical value for preventing and treating thrombosis.更多还原