摘要
新鲜猪心组织制成丙酮粉后,用0.45mol/L,pH4.2醋酸钾抽提组织型纤溶酶原活化物(t-PA)。抽提液经硫酸铵盐析,Benzamidine和血纤维蛋白亲和层析,Sephadex G-150凝胶过滤,纯化得到t-PA。比活11000IU/mg,经SDS-聚丙烯酰胺凝胶电泳鉴定,分子量为67000。 本文比较了t-PA、高分子量尿激酶(H-UK)和低分子量尿激酶(L-UK)的热稳定性及抑制剂对它们的抑制作用。结果表明,抑制剂对H-UK的抑制作用最强,L-UK次之,t-PA最弱;三者的热稳定性相似。
Fresh pig heart tissues were homogenized and delipidized with cold acetone. The dry powder was extracted with 0.45mol/L potassium acetate, pH4.2, the extract was fractionated with ammonium sulfate, followed by Benzamidine-Sepharose 4B affinity chromatography, Fibrin-Sepharose 4B affinity chromatography, and gel filtration on Sephadex G-150 sequentially. Thus purified t-PA (tissue plasminogen activator) has a specific activity of 11 000IU/mg and a molecular weight of 67kD as determined by SDS-polyacrylamide gel electrophoresis. The heat stability of t-PA,H-UK (high molecular weight urokinase) ,and L-UK (low molecular weight urokinase) and their inhibition by their inhibitor (purified from human placenta) were also compared. The results indicated that their heat stability was similar, but their inhibition by the inhibitor was different, with H-UK the strongest, L-UK the middle, and t-PA the weakest.
关键词
纤溶酶原
活化物
抑制剂
提纯
Tissue plasminogen activator
plasminogen activator inhibitor
