人心肌肌凝蛋白轻链的分离提纯与鉴定研究

Purification and Identification of Human Cardiac Myosin Light Chains

参考采用Schoβler方法并加以改进,分别应用等电点沉淀法、快速蛋白质液相层析(FPLC)及SDS聚丙烯酰胺凝胶制备电泳三种方法分离提纯肌凝蛋白轻链(CMLC),其各具有不同的优缺点,并对CMLC做了鉴定。同时证明在-20℃条件下,冻存时间较长或蛋白浓度较低的CMLC容易降解。

Normal human left ventricular myocardium was obtained within 5 hours after death. Crude cardiac myosin was extracted from the myocardium by the method of Schoβler, and cardiac myosin light chains (CMLC) were purified by isoelectric precipitation, fast protein liquid chromatography and preparative SDS-PAGE, respectively. The three methods were compared.The Ca2+-activated ATPase activity of cardiac myosin was 101.4 nmole Pi/mg/min. Molecular weights of CMLCI (24500) and CMLC Ⅱ (20000) were identified by analytical SDS-PAGE. All of the results mentioned above were consistent with data reported in the literature. An analytical electrophoresis test of the stability of CMLC stored at-20℃ demonstrated that liquid CMLC at long storage times or lower concentrations was easily degraded.