论文部分内容阅读
细胞的生化过程大都是由蛋白复合物完成的,研究蛋白复合物亚基的组成对于了解蛋白质的结构和生物学功能具有重要的意义,然而如何准确确定蛋白复合物中蛋白质亚基的数量(stoichiometry)仍然是一个挑战.近年来,活细胞体系单分子荧光成像技术的不断发展为原位实时动态地研究蛋白质的结构和性质提供了新的手段.本文主要介绍了应用活细胞全内反射单分子荧光成像技术表征细胞膜区蛋白复合物组成的3种方法,包括单分子漂白步数分析、荧光强度统计分布以及蛋白运动分析,并结合其基本原理介绍了这几种方法在活细胞体系膜蛋白研究中的应用.
The biochemical processes of cells are mostly done by protein complexes. Studying the composition of protein complex subunits is of great significance for understanding the structure and biological functions of proteins. However, how to accurately determine the number of protein subunits (stoichiometry ) Is still a challenge.In recent years, the continuous development of single molecule fluorescence imaging technology of living cell system provides a new means to study the structure and properties of protein in situ dynamically in real time.In this paper, Fluorescence imaging technology to characterize the composition of membrane protein complexes in the cell membrane by three methods, including single-molecule bleaching step number, statistical distribution of fluorescence intensity and protein kinetic analysis, combined with its basic principles of these methods introduced in living cell membrane protein research In the application.