Kyle J. Roux, Dae In Kim, Brian Burke
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引用次数: 201
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Abstract
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. Curr. Protoc. Protein Sci . 74:19.23.1-19.23.14. © 2013 by John Wiley & Sons, Inc.
BioID:蛋白质-蛋白质相互作用的筛选
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。该技术利用混杂生物素连接酶与生物素酸蛋白的接近性。连接酶与感兴趣的蛋白质融合并在细胞中表达,在细胞中对近端内源性蛋白质进行生物素化。由于这在自然界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,使它们能够通过标准生物素亲和力捕获进行选择性分离和鉴定。BioID鉴定的蛋白是目标蛋白的候选相互作用物。BioID可以应用于不溶性蛋白质,可以识别弱和/或瞬态相互作用,并且可以接受时间调节。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具有潜在的应用前景。咕咕叫。Protoc。蛋白质科学,74:19.23.1-19.23.14。©2013 by John Wiley &儿子,Inc。
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