Native-PAGE检测线粒体复合体酶I活性实验设计  

Practical design of native polyacrylamide gel electrophoresis for detection of mitochondrial complex enzyme I activity

作　　者：李莲[1] 王居锋 赖德华[1] LI Lian;WANG Jufeng;LAI Dehua(School of Life Sciences,Sun Yat-Sen University,Guangzhou 510275,China)

机构地区：[1]中山大学生命科学学院,广东广州510275

出　　处：《实验室科学》2025年第1期29-32,共4页Laboratory Science

基　　金：中山大学2022年度校级本科教学质量工程类项目(项目编号:教务[2022]91号)。

摘　　要：生物氧化是生物化学代谢部分的重要内容,因其复杂性和抽象性不易被学生理解。采用非变性聚丙烯酰胺凝胶电泳(Native polyacrylamide gel electrophoresis, Native-PAGE)构建了适用于本科实验教学的“线粒体复合体酶I活性方法”。从锥虫中提取线粒体粗体物,以10μg/孔样品蛋白上样量,进行非变性聚丙烯酰胺凝胶电泳, NADH脱氢酶酶促反应时间为30分钟,可得到分辨率高、条带清晰的特异性酶活性条带。将Native-PAGE方法应用到本科生化实验中,可有效拓展教学内容,有助于教学紧密联系科研,对本科生化实验教学的开展有借鉴意义。Biological oxidation is an important part of biochemical metabolism,which is difficult to be understood by students because of its complexity.In this paper,we established“the method for detection of mitochondrial complex enzyme I activity”by native polyacrylamide gel electrophoresis.The mitochondria were extracted from trypanosomes and proteins(10μg/well)were resolved on native polyacrylamide gels.Specific bands with high resolution and clear pattern were obtained after electrophoresis and 30 min reaction of NADH dehydrogenase.The experiment can effectively extend the content of teaching,correlated it with up-to-date scientific techniques,and could be referred in undergraduate Biochemistry practical teaching.

关 键 词：非变性聚丙烯酰胺凝胶电泳 生物氧化 线粒体复合体酶I 酶活性 

分 类 号：Q-33[生物学]