基于生物素连接酶的邻近标记技术在蛋白质组学中的研究进展

Research progress of proximity labeling technology based on biotin ligase in proteomics

基于生物素酶的蛋白质邻近标记技术是利用融合在感兴趣蛋白上的生物素连接酶对邻近的蛋白质生物素化,通过生物素与链酶亲和素之间的亲和力进行分离,再结合质谱分析鉴定出生物素化蛋白。该技术能够用于检测弱而短暂的蛋白质相互作用,也给在无膜细胞器和其他不易分离或纯化的亚细胞结构的上的蛋白互作研究提供了新的选择,很好地补充了传统研究蛋白质互作方法的空白。本文对近几年出现的基于生物素酶的蛋白质邻近标记的技术发展及其应用进行了综述。

Proximity-dependent biotinylation(PDB)uses biotin ligase fused to the protein of interest to bioti⁃nylate adjacent proteins,purify them with streptavidin beads,and then identify the biotinylated protein by mass spectrometry.This technology can be used to detect transient and/or low affinity interactions,provide a chance to learn more about membrane-less organelles and other subcellular structures that cannot be easily isolated or puri⁃fied,and fill the gap in traditional methods.This article summarizes the technological development and applica⁃tion of PDB in recent years.