CAJ | 학술논문

[目的]从蝰蛇(缅甸亚种)毒分离纯化一种凝血活性因子X激活物FVe-1,并研究其理化性质、序列测定以及凝血活性.[方法]应用阳离子交换层析、分子凝胶过滤层析分离纯化蛇毒,采用MALDI质谱测定法测定分子质量,等电聚焦电泳测定等电点,Edman降解法测定蛋白N末端氨基酸序列,发色底物法和SDS-PAGE等方法测定FVe-1的酶学特征和凝血活性.[结果]从蝰蛇(缅甸亚种)毒分离得到的F Ve-1为单体蛋白,相对分子质量为13 808,等电点4.6,0.5 mg FVe-1的凝血活性与1.5625 u的凝血酶或与54.93 ng RVVX的凝血活性相当;可激活FX,但对凝血酶和纤维蛋白原无作用；酶活性的适宜pH为6.5～ 7.5,适宜温度为25 ～ 60℃；为钙依赖性,凝血活性可被EDTA和DTT抑制;FVe-1的N端序列为NH2-N-L-Y-Q-F-G-E-M-I-N;具有呈剂量依赖性的促血浆凝固作用.[结论]从蝰蛇(缅甸亚种)毒纯化出一种F Ve-1是一个凝血因子X的激活物,可激活凝血因子X,但对凝血酶和纤维蛋白原的作用微弱.
[목적]종호사(면전아충)독분리순화일충응혈활성인자X격활물FVe-1,병연구기이화성질、서렬측정이급응혈활성.[방법]응용양리자교환층석、분자응효과려층석분리순화사독,채용MALDI질보측정법측정분자질량,등전취초전영측정등전점,Edman강해법측정단백N말단안기산서렬,발색저물법화SDS-PAGE등방법측정FVe-1적매학특정화응혈활성.[결과]종호사(면전아충)독분리득도적F Ve-1위단체단백,상대분자질량위13 808,등전점4.6,0.5 mg FVe-1적응혈활성여1.5625 u적응혈매혹여54.93 ng RVVX적응혈활성상당;가격활FX,단대응혈매화섬유단백원무작용；매활성적괄의pH위6.5～ 7.5,괄의온도위25 ～ 60℃；위개의뢰성,응혈활성가피EDTA화DTT억제;FVe-1적N단서렬위NH2-N-L-Y-Q-F-G-E-M-I-N;구유정제량의뢰성적촉혈장응고작용.[결론]종호사(면전아충)독순화출일충F Ve-1시일개응혈인자X적격활물,가격활응혈인자X,단대응혈매화섬유단백원적작용미약.
[Objective] To purify and characterize a novel factor X activator,Fve-1 from Daboia russelli siamensis (Myanmar) venom.[ Methods]F V e-1 was purified by ion-exchange chromatography and gel filtration.The hemostatic activity of F V e-1 was determined based on chromogenic substrates.The fibrinogen-clotting activity of F V e-1 was also determined.Thermal stability, pH stability,enzyme activity,and inhibition of F V e- 1 were determined by its remaining procoagulant activity.N-treminal sequence was determined by the method of automated Edman degradation.[ Results ]F V e-1 was achieved by chromatography with a molecular weight of 13,808 and an isoelectric point of 4.6. The hemostatic activity of 0.5 mg Fve-1 was equal to that of 1.5625 u thrombin or that of 54.93 ng RVV X. F V e-1 primarily activated F X, but did not affect on prothrombin and fibrinogen. The suitable pH and temperature range of F V e-1 was 6.5-7.5 and 25-60 ℃,respectively.The activity of F V e-1 was enhanced by Ca2+ and inhibited by EDTA and DTT.The N-terminal sequence of F V e-1 was NH2-N-L-Y-Q-F-G-E-M-I-N.[Conclusion] F V e-1 is a factor X-activating enzyme,which could activate FX to FX a,but have minimal effect on prothrombin and fibrinogen.