Drug Discovery Biology Theme, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville 3052, Victoria, Australia.
Hudson Institute of Medical Research, Clayton 3168, Victoria, Australia.
Science. 2021 Apr 9;372(6538). doi: 10.1126/science.abf7258. Epub 2021 Feb 18.
G protein-coupled receptors (GPCRs) are key regulators of information transmission between cells and organs. Despite this, we have only a limited understanding of the behavior of GPCRs in the apo state and the conformational changes upon agonist binding that lead to G protein recruitment and activation. We expressed and purified unmodified apo and peptide-bound calcitonin gene-related peptide (CGRP) receptors from insect cells to determine their cryo-electron microscopy (cryo-EM) structures, and we complemented these with analysis of protein conformational dynamics using hydrogen-deuterium exchange mass spectrometry and three-dimensional variance analysis of the cryo-EM data. Together with our previously published structure of the active, Gs-bound CGRP receptor complex, our work provides insight into the mechanisms of class B1 GPCR activation.
G 蛋白偶联受体（GPCRs）是细胞和器官之间信息传递的关键调节剂。尽管如此，我们对apo 状态下 GPCR 的行为以及激动剂结合导致 G 蛋白募集和激活的构象变化的了解仍然有限。我们从昆虫细胞中表达和纯化了未经修饰的 apo 和肽结合的降钙素基因相关肽（CGRP）受体，以确定它们的冷冻电镜（cryo-EM）结构，并使用氢氘交换质谱和 cryo-EM 数据的三维方差分析来补充这些结构的蛋白质构象动力学分析。结合我们之前发表的活性、Gs 结合的 CGRP 受体复合物的结构，我们的工作深入了解了 B1 类 GPCR 激活的机制。
