Varnaitė Renata, MacNeill Stuart A
School of Biology, University of St Andrews, North Haugh, St Andrews, Scotland, UK.
Proteomics. 2016 Oct;16(19):2503-2518. doi: 10.1002/pmic.201600123. Epub 2016 Jul 27.
Proximity-dependent biotin identification (BioID) is a recently developed method that allows the identification of proteins in the close vicinity of a protein of interest in living cells. BioID relies on fusion of the protein of interest with a mutant form of the biotin ligase enzyme BirA (BirA*) that is capable of promiscuously biotinylating proximal proteins irrespective of whether these interact directly or indirectly with the fusion protein or are merely located in the same subcellular neighborhood. The covalent addition of biotin allows the labeled proteins to be purified from cell extracts on the basis of their affinity for streptavidin and identified by mass spectrometry. To date, BioID has been successfully applied to study a variety of proteins and processes in mammalian cells and unicellular eukaryotes and has been shown to be particularly suited to the study of insoluble or inaccessible cellular structures and for detecting weak or transient protein associations. Here, we provide an introduction to BioID, together with a detailed summary of where and how the method has been applied to date, and briefly discuss technical aspects involved in the planning and execution of a BioID study.
邻近依赖性生物素识别（BioID）是一种最近开发的方法，可用于识别活细胞中目标蛋白附近的蛋白质。BioID依赖于将目标蛋白与生物素连接酶BirA的突变形式（BirA*）融合，该突变形式能够随机地对近端蛋白质进行生物素化，而不管这些蛋白质是直接还是间接与融合蛋白相互作用，或者仅仅位于同一亚细胞区域。生物素的共价添加使得标记的蛋白质能够基于它们对链霉亲和素的亲和力从细胞提取物中纯化出来，并通过质谱进行鉴定。迄今为止，BioID已成功应用于研究哺乳动物细胞和单细胞真核生物中的多种蛋白质和过程，并已被证明特别适合于研究不溶性或难以接近的细胞结构，以及检测弱的或短暂的蛋白质相互作用。在这里，我们介绍BioID，并详细总结该方法迄今为止的应用地点和方式，并简要讨论BioID研究规划和实施中涉及的技术方面。
