Daulat Avais, Maurice Pascal, Jockers Ralf
INSERM, U1016, Institut Cochin, Paris, France.
Methods Enzymol. 2013;521:329-45. doi: 10.1016/B978-0-12-391862-8.00018-1.
Biosynthesis and function of G protein-coupled receptors (GPCR) are accompanied by multiple GPCR-associated protein complexes. Despite considerable sequence diversity, all GPCRs are assumed to share a common 7-transmembrane-spanning architecture giving rise to an extracellular, intracellular, and transmembrane interface for the interaction with protein partners recognizing either linear or structural receptor epitopes. Different purification techniques have been developed in the past to identify GPCR-associated proteins other than classically known interacting proteins like heterotrimeric G proteins and β-arrestins. These techniques use either entire receptors or receptor subdomains as baits. We are presenting here two proteomic approaches developed in our laboratory to purify protein complexes interacting either with receptor subdomains from cell or tissue lysates or with entire receptors from intact cells.
G蛋白偶联受体（GPCR）的生物合成和功能伴随着多种与GPCR相关的蛋白复合物。尽管存在相当大的序列多样性，但所有GPCR都被认为具有共同的7次跨膜结构，从而形成用于与识别线性或结构性受体表位的蛋白伴侣相互作用的细胞外、细胞内和跨膜界面。过去已经开发出不同的纯化技术来鉴定除了经典已知的相互作用蛋白（如异源三聚体G蛋白和β-抑制蛋白）之外的与GPCR相关的蛋白。这些技术要么使用完整的受体，要么使用受体亚结构域作为诱饵。我们在此展示我们实验室开发的两种蛋白质组学方法，用于从细胞或组织裂解物中纯化与受体亚结构域相互作用的蛋白复合物，或从完整细胞中纯化与完整受体相互作用的蛋白复合物。
