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. 2023 Jan 23;14:376. doi: 10.1038/s41467-022-35726-z

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a Comparison of the orientation of Gαo in ACh-bound S1, ACh-bound S2, and Ixo-bound (PDB: 6OIK) states. b, c Comparison of αN and α5 helices in three structures, the α5 helix is shown in both side and bottom views. d Cartoon representation of the active (PDB: 6OIK) and inactive (PDB: 3UON) structures of the M2R with TM6 and the position of Met383^6.31 (red) highlighted. e HSQC spectra of Met383^6.31 in ACh-bound (cyan), ACh-GoA^GDP-bound (purple), and ACh-GoA^apyrase-bound (green) states. f HSQC spectra of Met383^6.31 in Ixo-bound (blue), Ixo-GoA^GDP-bound (brown), and Ixo-GoA^apyrase-bound (red) states. The apo-state spectrum of Met383^6.31 is shown in gray as a reference. g Comparison of the HSQC spectra of Met383^6.31 bound to ACh and Ixo along the process of GoA complex formation. The peak centers in ligand-bound spectra are shown as colored dots. h–j Alignment of the M1-G11 structure (PDB: 6OIJ) with the ACh-bound M2R-GoA S1 (h) and S2 (i) structures and the Ixo-bound M2R-GoA (PDB: 6OIK) structure (j). The different orientation between GoA and G11 is depicted with curved arrows. The clash between the extended TM5 helix with GoA is highlighted with dashed circle.