biostudies-literatureLiu QNational Natural Science Foundation of China4857https://www.ebi.ac.uk/biostudies/studies/S-EPMC7519161biostudies-literatureUnknown11(1)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 accNature communicationsDeSiphering receptor core-induced and ligand-dependent conformational changes in arrestin via genetic encoded trimethylsilyl ¹H-NMR probe.PMC7519161916401063170069221750003Zhang FWang QWNiu XGXiao KHJin CWLyu XZhu ZLLiu QGong ZLi ZJWang JHan MJYang ZYSun PChen XHuang SMYu XWang XYHe QTGuo SCKahsai AWSong SLYang FXiao PRuan KYang ZLin JYQu CXZhu KKLi FHXu ZKong WSun JPfalseDeSiphering receptor core-induced and ligand-dependent conformational changes in arrestin via genetic encoded trimethylsilyl ¹H-NMR probe.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 acc2020-01-01T00:00:00Z2020 Sep2026-04-30T05:18:42.55Z2020-10-29T09:32:36ZS-EPMC75191613297840210.1038/s41467-020-18433-5