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- Title
DeSiphering receptor core-induced and ligand-dependent conformational changes in arrestin via genetic encoded trimethylsilyl <sup>1</sup>H-NMR probe.
- Authors
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-jie
- Abstract
Characterization 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. Characterization of dynamic conformational changes in membrane protein complexes by NMR spectroscopy remains challenging. Here authors report the site-specific incorporation of 4-trimethylsilyl phenylalanine (TMSiPhe) into proteins, which enabled the characterization of multiple conformational states of a phospho-β2 adrenergic receptor/β-arrestin-1 complex in response to different receptor ligands.
- Subjects
ARRESTINS; MEMBRANE proteins; NUCLEAR magnetic resonance; GENETIC code; NUCLEAR magnetic resonance spectroscopy
- Publication
Nature Communications, 2020, Vol 11, Issue 1, pN.PAG
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-020-18433-5