ABSTRACT G-protein-coupled receptors (GPCRs) are seven-transmembrane proteins mediating cellular signals in response to extracellular stimuli. Although three-dimensional structures showcase

snapshots that can be sampled in the process and nuclear magnetic resonance detects conformational equilibria, the mechanism by which agonist-activated GPCRs interact with various effectors

remains elusive. Here, we used paramagnetic nuclear magnetic resonance for leucine amide resonances to visualize the structure of β2-adrenoreceptor in the full agonist-bound state, without

thermostabilizing mutations abolishing its activity. The structure exhibited a unique orientation of the intracellular half of the transmembrane helix 6, forming a cluster of

shed light on GPCR-mediated signal transduction. Access through your institution Buy or subscribe This is a preview of subscription content, access via your institution ACCESS OPTIONS Access

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customer support SIMILAR CONTENT BEING VIEWED BY OTHERS STRUCTURES OF Β1-ADRENERGIC RECEPTOR IN COMPLEX WITH GS AND LIGANDS OF DIFFERENT EFFICACIES Article Open access 14 July 2022 BINDING

KINETICS DRIVE G PROTEIN SUBTYPE SELECTIVITY AT THE Β1-ADRENERGIC RECEPTOR Article Open access 13 February 2024 MECHANISTIC INSIGHTS INTO G-PROTEIN COUPLING WITH AN AGONIST-BOUND

G-PROTEIN-COUPLED RECEPTOR Article 12 June 2024 DATA AVAILABILITY Atomic coordinates for β2AR-Δ in the fully activated state have been deposited in the PDB under accession code 6KR8. The NMR

data and restraints used in the structure calculations have been deposited in the Biological Magnetic Resonance Data Bank under accession number 36284. The other data that support the

findings of this study are available from the corresponding author upon reasonable request. CODE AVAILABILITY All code used in this study is available from the corresponding author upon

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  Download references ACKNOWLEDGEMENTS This work is supported by The Ministry of Education, Culture, Sports, Science and Technology and the Japan Society for the Promotion of Science KAKENHI

grant number JP17H06097 and by the development of innovative drug discovery technologies for middle-sized molecules from the Japan Agency for Medical Research and Development (to I.S.).

AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan Shunsuke Imai, Tomoki Yokomizo, Yutaka Kofuku, Yutaro

Shiraishi, Takumi Ueda & Ichio Shimada Authors * Shunsuke Imai View author publications You can also search for this author inPubMed Google Scholar * Tomoki Yokomizo View author

publications You can also search for this author inPubMed Google Scholar * Yutaka Kofuku View author publications You can also search for this author inPubMed Google Scholar * Yutaro

Shiraishi View author publications You can also search for this author inPubMed Google Scholar * Takumi Ueda View author publications You can also search for this author inPubMed Google

Scholar * Ichio Shimada View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS S.I. designed the study, constructed β2AR-Δ and its variants,

purified proteins, conducted GTP turnover assays with T.Y., acquired NMR spectra, analyzed the PRE data and calculated the PRE structure and wrote the manuscript. Y.K. established the

purification protocol of β2AR at the early stage of the project and constructed the plasmid for the expression of cystathionine-γ-synthase. Y.S. prepared the virus stock for the coexpression

of the Gs heterotrimer and cultured the cells by using the virus stock. T.U. performed the exchange Monte Carlo calculation. I.S. designed the study, analyzed the data and wrote the

manuscript. CORRESPONDING AUTHOR Correspondence to Ichio Shimada. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing interests. ADDITIONAL INFORMATION PUBLISHER’S NOTE

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION Supplementary Figs.

1–13 and Supplementary Table 1 REPORTING SUMMARY RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Imai, S., Yokomizo, T., Kofuku, Y. _et al._ Structural

equilibrium underlying ligand-dependent activation of β2-adrenoreceptor. _Nat Chem Biol_ 16, 430–439 (2020). https://doi.org/10.1038/s41589-019-0457-5 Download citation * Received: 13 July

2019 * Revised: 01 December 2019 * Accepted: 17 December 2019 * Published: 20 January 2020 * Issue Date: April 2020 * DOI: https://doi.org/10.1038/s41589-019-0457-5 SHARE THIS ARTICLE Anyone

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