Molecular Mechanism of Muscarinic Acetylcholine Receptor M3 Interaction with Gq

Overview

Journal Commun Biol

Specialty Biology

Date 2024 Mar 24

PMID 38521872

Authors

Donghee Ham

Asuka Inoue

Jun Xu

Yang Du

Ka Young Chung

Affiliations

Soon will be listed here.

Abstract

Muscarinic acetylcholine receptor M3 (M3) and its downstream effector Gq/11 are critical drug development targets due to their involvement in physiopathological processes. Although the structure of the M3-miniGq complex was recently published, the lack of information on the intracellular loop 3 (ICL3) of M3 and extensive modification of Gαq impedes the elucidation of the molecular mechanism of M3-Gq coupling under more physiological condition. Here, we describe the molecular mechanism underlying the dynamic interactions between full-length wild-type M3 and Gq using hydrogen-deuterium exchange mass spectrometry and NanoLuc Binary Technology-based cell systems. We propose a detailed analysis of M3-Gq coupling through examination of previously well-defined binding interfaces and neglected regions. Our findings suggest potential binding interfaces between M3 and Gq in pre-assembled and functionally active complexes. Furthermore, M3 ICL3 negatively affected M3-Gq coupling, and the Gαq AHD underwent unique conformational changes during M3-Gq coupling.

References

Westfield G, Rasmussen S, Su M, Dutta S, DeVree B, Chung K . Structural flexibility of the G alpha s alpha-helical domain in the beta2-adrenoceptor Gs complex. Proc Natl Acad Sci U S A. 2011; 108(38):16086-91. PMC: 3179071. DOI: 10.1073/pnas.1113645108. View

Okashah N, Wan Q, Ghosh S, Sandhu M, Inoue A, Vaidehi N . Variable G protein determinants of GPCR coupling selectivity. Proc Natl Acad Sci U S A. 2019; 116(24):12054-12059. PMC: 6575158. DOI: 10.1073/pnas.1905993116. View

Deutsch E, Csordas A, Sun Z, Jarnuczak A, Perez-Riverol Y, Ternent T . The ProteomeXchange consortium in 2017: supporting the cultural change in proteomics public data deposition. Nucleic Acids Res. 2016; 45(D1):D1100-D1106. PMC: 5210636. DOI: 10.1093/nar/gkw936. View

Sandhu M, Touma A, Dysthe M, Sadler F, Sivaramakrishnan S, Vaidehi N . Conformational plasticity of the intracellular cavity of GPCR-G-protein complexes leads to G-protein promiscuity and selectivity. Proc Natl Acad Sci U S A. 2019; 116(24):11956-11965. PMC: 6575595. DOI: 10.1073/pnas.1820944116. View

Sadler F, Ma N, Ritt M, Sharma Y, Vaidehi N, Sivaramakrishnan S . Autoregulation of GPCR signalling through the third intracellular loop. Nature. 2023; 615(7953):734-741. PMC: 10033409. DOI: 10.1038/s41586-023-05789-z. View

Wales T, Engen J . Hydrogen exchange mass spectrometry for the analysis of protein dynamics. Mass Spectrom Rev. 2005; 25(1):158-70. DOI: 10.1002/mas.20064. View

Zhang S, Gumpper R, Huang X, Liu Y, Krumm B, Cao C . Molecular basis for selective activation of DREADD-based chemogenetics. Nature. 2022; 612(7939):354-362. DOI: 10.1038/s41586-022-05489-0. View

Xu J, Wang Q, Hubner H, Hu Y, Niu X, Wang H . Structural and dynamic insights into supra-physiological activation and allosteric modulation of a muscarinic acetylcholine receptor. Nat Commun. 2023; 14(1):376. PMC: 9870890. DOI: 10.1038/s41467-022-35726-z. View

Ma X, Hu Y, Batebi H, Heng J, Xu J, Liu X . Analysis of βAR-G and βAR-G complex formation by NMR spectroscopy. Proc Natl Acad Sci U S A. 2020; 117(37):23096-23105. PMC: 7502740. DOI: 10.1073/pnas.2009786117. View

10.

Fukuda K, Kubo T, AKIBA I, Maeda A, Mishina M, Numa S . Molecular distinction between muscarinic acetylcholine receptor subtypes. Nature. 1987; 327(6123):623-5. DOI: 10.1038/327623a0. View

11.

Glukhova A, Draper-Joyce C, Sunahara R, Christopoulos A, Wootten D, Sexton P . Rules of Engagement: GPCRs and G Proteins. ACS Pharmacol Transl Sci. 2020; 1(2):73-83. PMC: 7089011. DOI: 10.1021/acsptsci.8b00026. View

12.

Weis D, Wales T, Engen J, Hotchko M, Ten Eyck L . Identification and characterization of EX1 kinetics in H/D exchange mass spectrometry by peak width analysis. J Am Soc Mass Spectrom. 2006; 17(11):1498-1509. DOI: 10.1016/j.jasms.2006.05.014. View

13.

Wess J, Bonner T, Dorje F, Brann M . Delineation of muscarinic receptor domains conferring selectivity of coupling to guanine nucleotide-binding proteins and second messengers. Mol Pharmacol. 1990; 38(4):517-23. View

14.

Maeda S, Qu Q, Robertson M, Skiniotis G, Kobilka B . Structures of the M1 and M2 muscarinic acetylcholine receptor/G-protein complexes. Science. 2019; 364(6440):552-557. PMC: 7034192. DOI: 10.1126/science.aaw5188. View

15.

Rose A, Elgeti M, Zachariae U, Grubmuller H, Hofmann K, Scheerer P . Position of transmembrane helix 6 determines receptor G protein coupling specificity. J Am Chem Soc. 2014; 136(32):11244-7. DOI: 10.1021/ja5055109. View

16.

Sperry J, Ryan Z, Kumar R, Gross M . Hydrogen/Deuterium Exchange Reflects Binding of Human Centrin 2 to Ca(2+) and Xeroderma Pigmentosum Group C Peptide: An Example of EX1 Kinetics. Int J Mass Spectrom. 2013; 330-332:302-309. PMC: 3578700. DOI: 10.1016/j.ijms.2012.10.013. View

17.

Tontson L, Babina A, Vosumaa T, Kopanchuk S, Rinken A . Characterization of heterotrimeric nucleotide-depleted Gα(i)-proteins by Bodipy-FL-GTPγS fluorescence anisotropy. Arch Biochem Biophys. 2012; 524(2):93-8. DOI: 10.1016/j.abb.2012.05.017. View

18.

Englander S, Kallenbach N . Hydrogen exchange and structural dynamics of proteins and nucleic acids. Q Rev Biophys. 1983; 16(4):521-655. DOI: 10.1017/s0033583500005217. View

19.

Wess J . Mutational analysis of muscarinic acetylcholine receptors: structural basis of ligand/receptor/G protein interactions. Life Sci. 1993; 53(19):1447-63. DOI: 10.1016/0024-3205(93)90618-d. View

20.

Morgan C, Hebling C, Rand K, Stafford D, Jorgenson J, Engen J . Conformational transitions in the membrane scaffold protein of phospholipid bilayer nanodiscs. Mol Cell Proteomics. 2011; 10(9):M111.010876. PMC: 3186204. DOI: 10.1074/mcp.M111.010876. View