Roux Kyle J, Kim Dae In, Burke Brian
Children's Health Research Center, Sanford Research, North Sioux Falls, South Dakota.
Department of Pediatrics, Sanford School of Medicine, University of South Dakota, Sioux Falls, South Dakota.
Curr Protoc Protein Sci. 2013 Nov 5;74:19.23.1-19.23.14. doi: 10.1002/0471140864.ps1923s74.
BioID is a unique method to screen for physiologically relevant protein interactions that occur in living cells. This technique harnesses a promiscuous biotin ligase to biotinylate proteins based on proximity. The ligase is fused to a protein of interest and expressed in cells, where it biotinylates proximal endogenous proteins. Because it is a rare protein modification in nature, biotinylation of these endogenous proteins by BioID fusion proteins enables their selective isolation and identification with standard biotin-affinity capture. Proteins identified by BioID are candidate interactors for the protein of interest. BioID can be applied to insoluble proteins, can identify weak and/or transient interactions, and is amenable to temporal regulation. Initially applied to mammalian cells, BioID has potential application in a variety of cell types from diverse species.
生物识别（BioID）是一种筛选活细胞中发生的生理相关蛋白质相互作用的独特方法。该技术利用一种混杂的生物素连接酶，根据接近程度对蛋白质进行生物素化。连接酶与感兴趣的蛋白质融合并在细胞中表达，在细胞中它会对近端内源性蛋白质进行生物素化。由于生物素化在自然界中是一种罕见的蛋白质修饰，通过BioID融合蛋白对这些内源性蛋白质进行生物素化，能够使用标准的生物素亲和捕获方法对它们进行选择性分离和鉴定。通过BioID鉴定的蛋白质是感兴趣蛋白质的候选相互作用蛋白。BioID可应用于不溶性蛋白质，能识别弱相互作用和/或瞬时相互作用，并且易于进行时间调控。BioID最初应用于哺乳动物细胞，在来自不同物种的多种细胞类型中都具有潜在应用价值。
