肌钙蛋白Ⅰ提取纯化新方法的研究  

Studies on a New Method of Extraction and Purification of Troponin Ⅰ

作　　者：刘忠英[1] 胡秀丽[1] 石毅[1] 姜熙罗[1] 安汝国[1] 马学华[2] 

机构地区：[1]吉林大学生物工程研究所,长春130021 [2]哈尔滨医科大学附属第一临床医院检验科

出　　处：《高等学校化学学报》2002年第5期818-821,共4页Chemical Journal of Chinese Universities

基　　金：吉林省自然科学基金(批准号:990324)资助.

摘　　要：兔骨骼肌匀浆液经离心后，依次经DEAE－Sephadex A25，Sephadex G75及Butyl Sepharose F．F柱层析，得到了兔骨骼肌肌钙蛋白C（sTnC）纯品．将其与Sepharose 4B凝胶偶联，制成sTnC亲和层析凝胶．人心肌或骨骼肌经匀浆、离心后上sTnC亲和层析柱，一步分离可获得人心肌肌钙蛋白Ⅰ（cTn I）或人骨骼肌肌钙蛋白Ⅰ（sTn I）纯品．HPLC分析出现单一峰．SDS－PAGE分析得到一条电泳带，其分子量分别为25 000及 21 000．用 20 g sTnC亲和凝胶处理 40 g人心肌时，相当于每 100 g心肌可得到 82.6 mg cTnI.Abstract Rabbit skeletal muscle was homogenized and centrifuged, then applied to DEAE-Sephadex A25, Sephadex G75 and Butyl Sepharose columns in turn. Finally, the purified sTnC was obtained. The affinity chromatography gel was prepared by binding the purified sTnC onto the CNBr-act Sepharose 4B. Troponin I was isolated directly from whole skeletal and cardial muscle by using the affinity chromatography gel. Both the results of HPLC and SDS-PAGE show a single band. The molecular weights of the purified sTn I and cTn I were 21 000 and 25 000, respectively. The yield of sTnC was 82. 6 mg/100 g cardiac tissue, which was higher than that obtained by using the previous method. In this work, 91. 6 mg purified cTn I were obtained.

关 键 词：肌钙蛋白C 提取 心肌肌钙蛋白Ⅰ 骨骼肌肌钙蛋白Ⅰ 急性心肌梗塞 临床诊断 

分 类 号：R542.22[医药卫生—心血管疾病]