Carpenter Byron, Nehmé Rony, Warne Tony, Leslie Andrew G W, Tate Christopher G
Nature. 2016 Aug 4;536(7614):104-7. doi: 10.1038/nature18966. Epub 2016 Jul 27.
G-protein-coupled receptors (GPCRs) are essential components of the signalling network throughout the body. To understand the molecular mechanism of G-protein-mediated signalling, solved structures of receptors in inactive conformations and in the active conformation coupled to a G protein are necessary. Here we present the structure of the adenosine A(2A) receptor (A(2A)R) bound to an engineered G protein, mini-Gs, at 3.4 Å resolution. Mini-Gs binds to A(2A)R through an extensive interface (1,048 Å2) that is similar, but not identical, to the interface between Gs and the β2-adrenergic receptor. The transition of the receptor from an agonist-bound active-intermediate state to an active G-protein-bound state is characterized by a 14 Å shift of the cytoplasmic end of transmembrane helix 6 (H6) away from the receptor core, slight changes in the positions of the cytoplasmic ends of H5 and H7 and rotamer changes of the amino acid side chains Arg3.50, Tyr5.58 and Tyr7.53. There are no substantial differences in the extracellular half of the receptor around the ligand binding pocket. The A(2A)R-mini-Gs structure highlights both the diversity and similarity in G-protein coupling to GPCRs and hints at the potential complexity of the molecular basis for G-protein specificity.
G蛋白偶联受体(GPCRs)是遍布全身的信号网络的重要组成部分。为了理解G蛋白介导的信号传导的分子机制,需要解析处于非活性构象以及与G蛋白偶联的活性构象的受体结构。在此,我们展示了与工程化G蛋白mini-Gs结合的腺苷A2A受体(A2AR)的结构,分辨率为3.4Å。Mini-Gs通过一个广泛的界面(1048Å2)与A2AR结合,该界面与Gs和β2肾上腺素能受体之间的界面相似但不完全相同。受体从激动剂结合的活性中间状态转变为与活性G蛋白结合的状态的特征是,跨膜螺旋6(H6)的胞质端从受体核心向外移动14Å,H5和H7的胞质端位置略有变化,以及氨基酸侧链Arg3.50、Tyr5.58和Tyr7.53的旋转异构体变化。在配体结合口袋周围的受体细胞外部分没有实质性差异。A2AR-mini-Gs结构突出了G蛋白与GPCRs偶联的多样性和相似性,并暗示了G蛋白特异性分子基础的潜在复杂性。