双杂交和质谱技术在蛋白质-蛋白质相互作用研究中的应用

基因的功能是由蛋白质来执行的,而蛋白质要通过与其他生物分子相互作用来完成其各种生物功能。因此,如果能够快速做出蛋白质在不同时间、空间和不同环境中的相互作用图谱,就会帮助我们了解这些蛋白质的功能,进而了解许多生命活动的机制。目前,用于大规模研究蛋白质间相互作用的方法主要有酵母双杂交系统及其衍生系统、亲和纯化与质谱分析联用技术,前者用于研究蛋白分子间的两两相互作用,后者用于研究蛋白质复合物间的相互作用。本文主要阐述了酵母双杂交、细菌双杂交、哺乳动物细胞双杂交、亲和纯化与质谱联用技术在大规模蛋白质相互作用研究中的应用。

Applications of Two-Hybrid Systems and Mass Spectrometry to Investigate Protein-Protein Interactions

Genes usually ascribe their function through proteins, and the function of a protein is to interact with other molecules. Therefore, if one could instantly map these interacting molecules in space, time and milieu, then one would be able to understand the function of proteins. Now, the approaches for high-throughput screening and identifying protein-protein interaction mainly are the yeast two-hybrid system and its derivatives for binary interactions, and systematic affinity purification followed by mass spectrometry for complex interactions. In this paper, applications of yeast, bacterial and mammalian cell two-hybrid systems, and the affinity purification followed by mass spectrometry to investigate protein-protein interactions were reviewed.