DeSiphering receptor core-induced and ligand-dependent conformational changes in arrestin via genetic encoded trimethylsilyl 1H-NMR probe
-
Published:2020-09-25
Issue:1
Volume:11
Page:
-
ISSN:2041-1723
-
Container-title:Nature Communications
-
language:en
-
Short-container-title:Nat Commun
Author:
Liu Qi, He Qing-tao, Lyu Xiaoxuan, Yang Fan, Zhu Zhong-liang, Xiao Peng, Yang Zhao, Zhang Feng, Yang Zhao-ya, Wang Xiao-yan, Sun Peng, Wang Qian-wen, Qu Chang-xiu, Gong Zheng, Lin Jing-yu, Xu Zhen, Song Shao-le, Huang Shen-ming, Guo Sheng-chao, Han Ming-jieORCID, Zhu Kong-kai, Chen Xin, Kahsai Alem W., Xiao Kun-Hong, Kong WeiORCID, Li Fa-hui, Ruan KeORCID, Li Zi-jian, Yu Xiao, Niu Xiao-gang, Jin Chang-wen, Wang Jiangyun, Sun Jin-pengORCID
Abstract
AbstractCharacterization of the dynamic conformational changes in membrane protein signaling complexes by nuclear magnetic resonance (NMR) spectroscopy remains challenging. Here we report the site-specific incorporation of 4-trimethylsilyl phenylalanine (TMSiPhe) into proteins, through genetic code expansion. Crystallographic analysis revealed structural changes that reshaped the TMSiPhe-specific amino-acyl tRNA synthetase active site to selectively accommodate the trimethylsilyl (TMSi) group. The unique up-field 1H-NMR chemical shift and the highly efficient incorporation of TMSiPhe enabled the characterization of multiple conformational states of a phospho-β2 adrenergic receptor/β-arrestin-1(β-arr1) membrane protein signaling complex, using only 5 μM protein and 20 min of spectrum accumulation time. We further showed that extracellular ligands induced conformational changes located in the polar core or ERK interaction site of β-arr1 via direct receptor transmembrane core interactions. These observations provided direct delineation and key mechanism insights that multiple receptor ligands were able to induce distinct functionally relevant conformational changes of arrestin.
Funder
National Natural Science Foundation of China
Publisher
Springer Science and Business Media LLC
Subject
General Physics and Astronomy,General Biochemistry, Genetics and Molecular Biology,General Chemistry
Reference65 articles.
1. Weis, W. I. & Kobilka, B. K. The molecular basis of G protein-coupled receptor activation. Annu Rev. Biochem. 87, 897–919 (2018). 2. Yan, N. Structural biology of the major facilitator superfamily transporters. Annu Rev. Biophys. 44, 257–283 (2015). 3. Zhou, X. E. et al. Identification of phosphorylation codes for arrestin recruitment by G protein-coupled receptors. Cell 170, 457–469 e413 (2017). 4. Gregorio, G. G. et al. Single-molecule analysis of ligand efficacy in β2AR-G-protein activation. Nature 547, 68–73 (2017). 5. Kaplan, M., Pinto, C., Houben, K. & Baldus, M. Nuclear magnetic resonance (NMR) applied to membrane-protein complexes. Q. Rev. Biophys. 49, e15 (2016).
Cited by
25 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献
|
|