Department of Chemistry, University of Florida, Gainesville, FL 32611, USA.
Department of Chemistry, University of Florida, Gainesville, FL 32611, USA.
Methods. 2020 Aug 1;180:79-88. doi: 10.1016/j.ymeth.2020.08.008. Epub 2020 Sep 8.
G protein-coupled receptors (GPCRs) represent the largest class of "druggable" proteins in the human genome. For more than a decade, crystal structures and, more recently, cryoEM structures of GPCR complexes have provided unprecedented insight into GPCR drug binding and cell signaling. Nevertheless, structure determination of receptors in complexes with weakly binding molecules or complex polypeptides remains especially challenging, including for hormones, many of which have so far eluded researchers. Nuclear magnetic resonance (NMR) spectroscopy has emerged as a promising approach to determine structures of ligands bound to their receptors and to provide insights into the dynamics of GPCR-bound drugs. The capability to investigate compounds with weak binding affinities has also been leveraged in NMR applications to identify novel lead compounds in drug screening campaigns. We review recent structural biology studies of GPCR ligands by NMR, highlighting new methodologies enabling studies of GPCRs with native sequences and in native-like membrane environments that provide insights into important drugs and endogenous ligands.
G 蛋白偶联受体 (GPCRs) 是人类基因组中最大的“可成药”蛋白类群。十多年来,GPCR 复合物的晶体结构,以及最近的冷冻电镜结构,为 GPCR 药物结合和细胞信号转导提供了前所未有的深入见解。然而,与弱结合分子或复杂多肽结合的受体的结构测定仍然特别具有挑战性,其中包括许多激素,迄今为止,这些激素仍让研究人员感到困惑。核磁共振 (NMR) 光谱已成为一种很有前途的方法,可以确定配体与受体结合的结构,并深入了解与 GPCR 结合的药物的动力学。NMR 应用中的这种能够研究弱结合亲和力化合物的能力也被用于药物筛选活动中,以鉴定新的先导化合物。我们综述了通过 NMR 研究 GPCR 配体的最新结构生物学研究,重点介绍了能够研究具有天然序列和天然样膜环境的 GPCR 的新方法学,这些方法学为重要药物和内源性配体提供了深入见解。