2. Academic Validation
  3. DeSiphering receptor core-induced and ligand-dependent conformational changes in arrestin via genetic encoded trimethylsilyl 1H-NMR probe

DeSiphering receptor core-induced and ligand-dependent conformational changes in arrestin via genetic encoded trimethylsilyl 1H-NMR probe

  • Nat Commun. 2020 Sep 25;11(1):4857. doi: 10.1038/s41467-020-18433-5.
Qi Liu  # 1 2,  Qing-Tao He  # 1 3 4,  Xiaoxuan Lyu  # 1,  Fan Yang  # 2 4,  Zhong-Liang Zhu  # 5,  Peng Xiao 3 4,  Zhao Yang 2 4,  Feng Zhang 1,  Zhao-Ya Yang 1 3,  Xiao-Yan Wang 1,  Peng Sun 6,  Qian-Wen Wang 6,  Chang-Xiu Qu 3 4,  Zheng Gong 3,  Jing-Yu Lin 2,  Zhen Xu 1,  Shao-le Song 1,  Shen-Ming Huang 4,  Sheng-Chao Guo 3 4,  Ming-Jie Han 1 7,  Kong-Kai Zhu 8,  Xin Chen 9,  Alem W Kahsai 10,  Kun-Hong Xiao 11,  Wei Kong 4,  Fa-Hui Li 1,  Ke Ruan 12,  Zi-Jian Li 4,  Xiao Yu 2,  Xiao-Gang Niu 13,  Chang-Wen Jin 13,  Jiangyun Wang 14 15,  Jin-Peng Sun 16 17
Affiliations

Affiliations

  • 1 Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang district, Beijing, 100101, China.
  • 2 Key Laboratory Experimental Teratology of the Ministry of Education and Department of Physiology, School of Basic Medical Sciences, Cheeloo college of Medicine, Shandong University, 44 Wenhua Xi Road, Jinan, Shandong, 250012, China.
  • 3 Key Laboratory Experimental Teratology of the Ministry of Education and Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Cheeloo college of Medicine, Shandong University, 44 Wenhua Xi Road, Jinan, 250012, Shandong, China.
  • 4 Key Laboratory of Molecular Cardiovascular Science, Ministry of Education, Peking University, 15 Xueyuan Road, Haidian District, Beijing, 100191, China.
  • 5 School of Life Sciences, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui, 230026, China.
  • 6 Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, 30 Xiaohongshan Road, Wuchang District, Wuhan, Hubei, 430071, China.
  • 7 Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 Xiqi Road, Airport Economic Zone, Dongli District, Tianjin, 300308, China.
  • 8 School of Biological Science and Technology, University of Jinan, 336 Nanxinzhuangxi Road, Shizhong District, Jinan, 250022, China.
  • 9 Department of Medicinal Chemistry, School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu, 213164, China.
  • 10 Duke University, School of Medicine, Durham, NC, 27705, USA.
  • 11 Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
  • 12 Hefei National Laboratory for Physical Science at the Microscale, University of Science and Technology of China, 443 Huangshan Road, Hefei, Anhui, 230027, China.
  • 13 Beijing Nuclear Magnetic Resonance Center, College of Chemistry and Molecular Engineering, School of Life Sciences, Peking University, Beijing, 100084, China.
  • 14 Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang district, Beijing, 100101, China. [email protected].
  • 15 College of Life Sciences and School of Future Technology, University of Chinese Academy of Sciences, Beijing, 100049, China. [email protected].
  • 16 Key Laboratory Experimental Teratology of the Ministry of Education and Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Cheeloo college of Medicine, Shandong University, 44 Wenhua Xi Road, Jinan, 250012, Shandong, China. [email protected].
  • 17 Key Laboratory of Molecular Cardiovascular Science, Ministry of Education, Peking University, 15 Xueyuan Road, Haidian District, Beijing, 100191, China. [email protected].
  • # Contributed equally.
PMID: 32978402 DOI: 10.1038/s41467-020-18433-5
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.

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