Le Sage Valerie, Cinti Alessandro, Mouland Andrew J
HIV-1 RNA Trafficking Laboratory, Lady Davis Institute at the Jewish General Hospital, Montréal, Québec, Canada.
Department of Medicine, McGill University, Montréal, Québec, Canada.
Curr Protoc Cell Biol. 2016 Dec 1;73:17.19.1-17.19.12. doi: 10.1002/cpcb.11.
Complex interaction networks orchestrate key cellular processes including but not limited to transcription, translation, metabolism, and cell signaling. Delineating these interactions will aid in deciphering the regulation and function of these pathways and potential for manipulation. Proximity-dependent biotin identification (BioID) is quickly gaining popularity as a powerful tool for identifying novel protein-protein and proximity-based interactions in live cells. This technique relies on a promiscuous biotin ligase, which is fused to a protein of interest and, upon expression in the desired cell, will biotinylate proximal endogenous proteins. In vivo protein-protein interactions can be very transient and occur momentarily to facilitate signaling or a metabolic function. BioID is useful in identifying these weak and/or transient interactions that are not detected by traditional methods such as yeast two-hybrid or affinity purification. Here, we outline a BioID protocol that can be used as a workflow to guide a new application. © 2016 by John Wiley & Sons, Inc.
复杂的相互作用网络协调着关键的细胞过程，包括但不限于转录、翻译、代谢和细胞信号传导。描绘这些相互作用将有助于解读这些途径的调控和功能以及操纵的潜力。邻近依赖性生物素识别（BioID）作为一种在活细胞中识别新型蛋白质-蛋白质和基于邻近性的相互作用的强大工具，正迅速受到欢迎。该技术依赖于一种混杂的生物素连接酶，它与感兴趣的蛋白质融合，在所需细胞中表达时，会将近端内源性蛋白质生物素化。体内蛋白质-蛋白质相互作用可能非常短暂，瞬间发生以促进信号传导或代谢功能。BioID可用于识别传统方法（如酵母双杂交或亲和纯化）无法检测到的这些弱和/或短暂的相互作用。在这里，我们概述了一种BioID方案，可作为指导新应用的工作流程。© 2016 John Wiley & Sons, Inc.
