Department of Biochemistry & Molecular Biology, Dalhousie University, 5850 College Street, Halifax, Nova Scotia, Canada.
Biochem Cell Biol. 2011 Apr;89(2):98-105. doi: 10.1139/o10-142.
G-protein coupled receptors (GPCRs) are ubiquitous membrane proteins allowing intracellular responses to extracellular factors that range from photons of light to small molecules to proteins. Despite extensive exploitation of GPCRs as therapeutic targets, biophysical characterization of GPCR-ligand interactions remains challenging. In this minireview, we focus on techniques that have been successfully used for structural and biophysical characterization of peptide ligands binding to their cognate GPCRs. The techniques reviewed include solution-state nuclear magnetic resonance (NMR) spectroscopy, solid-state NMR, X-ray diffraction, fluorescence spectroscopy and single-molecule fluorescence methods, flow cytometry, surface plasmon resonance, isothermal titration calorimetry, and atomic force microscopy. The goal herein is to provide a cohesive starting point to allow selection of techniques appropriate to the elucidation of a given GPCR-peptide interaction.
G 蛋白偶联受体 (GPCRs) 是普遍存在的膜蛋白，允许细胞内对从光量子到小分子再到蛋白质等各种细胞外因子做出反应。尽管 GPCR 已被广泛用作治疗靶点，但对 GPCR-配体相互作用的生物物理特性的研究仍然具有挑战性。在本篇简评中，我们重点介绍了成功用于结构和生物物理特性研究的技术，这些技术用于研究与 GPCR 相互作用的肽配体。所综述的技术包括溶液态核磁共振 (NMR) 光谱学、固态 NMR、X 射线衍射、荧光光谱学和单分子荧光方法、流式细胞术、表面等离子体共振、等温滴定量热法和原子力显微镜。本文的目的是提供一个连贯的起点，以便选择合适的技术来阐明特定的 GPCR-肽相互作用。
