用于鉴定哺乳动物细胞中蛋白质-蛋白质相互作用的位点特异性光交联蛋白质组学方案。
Protocol for Site-Specific Photo-Crosslinking Proteomics to Identify Protein-Protein Interactions in Mammalian Cells.
机构信息
State Key Laboratory of Chemical Oncogenomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen 518055, China.
出版信息
STAR Protoc. 2020 Sep 15;1(3):100109. doi: 10.1016/j.xpro.2020.100109. eCollection 2020 Dec 18.
Protein-protein interactions (PPIs) play essential roles in almost all aspects of cellular processes. However, PPIs remain challenging to study due to their substoichiometry, low affinity, dynamic nature, and context dependence. Here, we present a protocol for the capture and identification of PPIs in live mammalian cells, which relies on site-specific photo-crosslinking in live cells, affinity purification, and quantitative proteomics. The protocol facilitates efficient and reliable identification of the interacting proteins of a given protein of interest in live cells. For complete details on the use and execution of this protocol, please refer to Wu et al. (2020).
蛋白质-蛋白质相互作用 (PPIs) 在几乎所有细胞过程中都发挥着重要作用。然而，由于它们的亚化学计量、低亲和力、动态性质和上下文依赖性，PPIs 的研究仍然具有挑战性。在这里，我们介绍了一种在活哺乳动物细胞中捕获和鉴定蛋白质-蛋白质相互作用的方案，该方案依赖于活细胞中的特异性光交联、亲和纯化和定量蛋白质组学。该方案有助于在活细胞中有效且可靠地鉴定给定感兴趣蛋白质的相互作用蛋白。有关此方案的使用和执行的完整详细信息，请参阅 Wu 等人。(2020)。
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