基于液质联用技术的蛋白质-蛋白质相互作用研究进展

Research Progress of Protein-Protein Interaction Based on Liquid Chromatography Mass Spectrometry

蛋白质-蛋白质相互作用调控着细胞内众多生物过程,蛋白质间相互作用网络的绘制对解析复杂的生物过程至关重要.面对生物体中复杂的蛋白质-蛋白质相互作用,液质联用技术不仅具有灵敏度高的鉴定优势,还可以对数以千计的蛋白质进行定量分析.因此,对目标蛋白质进行富集、标记或共分级的处理后,结合液质联用技术对蛋白质准确而灵敏的鉴定,这类技术已被广泛应用于复杂样本中蛋白质-蛋白质相互作用网络的解析.目前基于液质联用技术的几种常用的方式,包括亲和纯化质谱方法(AP-MS)、近程标记质谱方法(PDB-MS)、化学交联质谱方法(XL-MS)和共分级偶联质谱方法(CF-MS)等.本综述讨论了这些方法的基本原理、优点和在细胞内解析蛋白质间相互作用的应用.

Protein-protein interactions are involved in the regulation of many biological processes in cells,and the mapping of protein-protein interaction networks is crucial for understanding complex biological processes.Liquid chromatography-mass spectrometry(LC-MS)can identify and quantify thousands of proteins simultaneously in complex organisms with its high sensitivity and accuracy.Therefore,after the enrichment,labeling or co-fractionation of target proteins,combined with LC-MS technology to identify proteins accurately and sensitively,such techniques have been widely used in the analysis of protein-protein interaction networks in the complex samples.There are LC-MS-based methods for studying protein-protein interactions,including affinity purification mass spectrometry(AP-MS),proximity-dependent biotinylation coupled to mass spectrometry(PDB-MS),chemical cross-linking with mass spectrometry(XL-MS)and co-fractionation mass spectrometry(CF-MS).This review discusses the mechanism,advantages and applications of these methods for the identification towards the protein-protein interactions in cells.