细胞质膜及其蛋白质复合物分离与定性方法研究  

Separation and analysis of protein complexes in plasma membrane

作　　者：胡凡[1] 许志洋[1] 夏峥嵘[1] 王富强[1] 王玲[1] 

机构地区：[1]南京医科大学分析测试中心,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2013年第3期407-411,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：南京医科大学科技发展基金面上项目(2011NJ-MU69;2011NJMU70)

摘　　要：目的:探讨细胞质膜及其蛋白质复合物分离与定性的方法。方法:结合差速离心法、密度梯度离心法、双水相法分离得到细胞质膜,并利用透射电镜和Western blot等实验手段验证分离所得细胞质膜的纯度。结果:本文采用了蓝绿温和凝胶电泳法对细胞质膜上的蛋白质复合物进行一维分离,得到23种蛋白质复合物;接着对23种蛋白质复合物进行第二相垂直电泳分离,分离出135种蛋白质;最后鉴定78种是质膜相关蛋白,其余57种来源内膜系统。结论:多种提纯方法的联用可以有效提高蛋白分离率与分离纯度,高纯度的质膜对质膜蛋白质组的研究具有重要意义。Objective：Discuss the methods of plasma membrane and its protein complex separation and qualitative. Methods：We evaluated the application of ＂density gradient ultracentfifugation＂and ＂aqueous two-phase partition＂to separate high-quality plasma membrane. Western blot and transmission electron microscope were used in the following research to validate the purity of the separated cell membrane. Results：We applied the blue-native method to process the one-dimensional separation of the protein complex in the cell membrane and 23 kinds of protein complex were separated. We also processed the second-dimensional separation of the 23 kinds of complex and identified 135 proteins, of which 78 were proteins correlated to plasma membrane and 57 were from endomembrane system. Conclusion：Our results show that the combination of different kinds of purification methods can effectively increase the protein separation rate and separation purity. High purity membrane on plasma membrane proteome research has important significance.

关 键 词：细胞质膜 BN-PAGE 蛋白质复合物 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]