新方法的开发对于获得对现有方法或技术无法获得的生物过程的进一步见解至关重要。蛋白质-蛋白质相互作用的检测通常具有挑战性,特别是对于弱和瞬时相互作用或膜蛋白。在过去的十年里,已经开发了几种邻近标记方法来探索活细胞中的蛋白质相互作用。其中,最有效的是基于蛋白质伴侣修饰,如生物素化或嘌呤化。这些技术是基于酶的工程变体,如过氧化物酶或释放反应分子的连接酶,在特定底物的存在下,结合周围的蛋白质。将感兴趣的蛋白质(POI)融合到这些酶中可以定义无偏的“proxisome”,“也就是说,所有与POI相互作用或紧邻的蛋白质。这里,描述了可用的不同的接近标签工具,并提供了全面的比较以讨论优点和局限性。
The development of new approaches is critical to gain further insights into biological processes that cannot be obtained by existing methods or technologies. The detection of protein-protein interaction is often challenging, especially for weak and transient interactions or for membrane proteins. Over the last decade, several proximity-tagging methodologies have been developed to explore protein interactions in living cells. Among those, the most efficient are based on protein partner modification, such as biotinylation or pupylation. Such technologies are based on engineered variants of enzymes like peroxidases or ligases that release reactive molecules, in the presence of specific substrates, that bind surrounding proteins. Fusing a protein of interest (POI) to these enzymes allows the definition of an unbiased \"
proxisome,\" that is, all of the proteins in interaction or in close vicinity of the POI. Here, the different proximity-labeling tools available are described and comprehensive comparison to discuss advantages and limitations is provided.