邻近标记在蛋白质组学中的发展及应用

The Proceedings and Applications of Proximity Labeling in Proteomics

人体内各种复杂的生命活动离不开蛋白质之间的相互作用。这种相互作用具有瞬时性和结合力弱等特点,并受到多种动态调节,特别是蛋白质翻译后修饰(post-translation modifications,PTM)。传统的亲和质谱检测方法存在蛋白纯化的局限性,在高效检测到动态变化方面存在不足。邻近标记是一种能够给与靶蛋白质瞬时靠近,或者互作(邻近)的蛋白质加上生物素的技术,它与质谱检测技术的联合使用能检测细胞过程中弱的、瞬时的蛋白质相互作用,有效解决上述问题。本文综述了基于生物素的邻近标记方法的发展现状,从依赖于融合序列的生物素标记开始,依次介绍有关生物素连接酶、过氧化物酶及其进化后的2代标记方法等经典生物素标记的方法和原理,比较各个方法间的差异和优缺点;也列举了一些近年来新出现的标记方法,如将生物素连接酶进行拆分、鉴定蛋白质在不同复合物中功能的方法、抗体靶向的标记方法,以及其他来源的生物素连接酶突变体,例如枯草芽孢杆菌(Bacillus subtilis)的C端氨基酸突变的生物素连接酶,能够应用在苍蝇和蠕虫中的生物素连接酶突变体。本文对这些方法进行归纳总结,旨在为初步接触该领域的科研工作者提供参考,同时也希望能够提供一些新的思路,推动蛋白质相互作用组学的发展。

The complexity of life activities in humans are inseparable from protein-protein interactions( PPIs). Most PPIs are dynamic,especially by numerous transient post-translational modifications( PTM). Proximity labeling is a recently developed technique,which labels the potential binding proteins of a target protein based on proximity by biotin. It can be used in combination with mass spectrometry to detect weak and transient PPIs in cellular processes. This paper reviews the development of different biotin-based proximity labeling methods. Starting from the biotin labeling method based on the fused substrate sequences,the principle of other labeling probes such as BirA*,an engineered peroxidase( APEX),Bacillus subtilis( BASU) and APEX2 are discussed. Methods developed in the past few years such as antibody-targeted proximity labeling and split BioID are also introduced. The application and prospect of these methods in investigating PPIs might inspire new researches in this field.