Pharmacological hallmarks of allostery at the M4 muscarinic receptor elucidated through structure and dynamics
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Abstract
Allosteric modulation of G protein-coupled receptors (GPCRs) is a major paradigm in drug discovery. Despite decades of research, a molecular-level understanding of the general principles that govern the myriad pharmacological effects exerted by GPCR allosteric modulators remains limited. The M 4 muscarinic acetylcholine receptor (M 4 mAChR) is a validated and clinically relevant allosteric drug target for several major psychiatric and cognitive disorders. In this study, we rigorously quantified the affinity, efficacy, and magnitude of modulation of two different positive allosteric modulators, LY2033298 (LY298) and VU0467154 (VU154), combined with the endogenous agonist acetylcholine (ACh) or the high-affinity agonist iperoxo (Ipx), at the human M 4 mAChR. By determining the cryo-electron microscopy structures of the M 4 mAChR, bound to a cognate G i1 protein and in complex with ACh, Ipx, LY298-Ipx, and VU154-Ipx, and applying molecular dynamics simulations, we determine key molecular mechanisms underlying allosteric pharmacology. In addition to delineating the contribution of spatially distinct binding sites on observed pharmacology, our findings also revealed a vital role for orthosteric and allosteric ligand–receptor–transducer complex stability, mediated by conformational dynamics between these sites, in the ultimate determination of affinity, efficacy, cooperativity, probe dependence, and species variability. There results provide a holistic framework for further GPCR mechanistic studies and can aid in the discovery and design of future allosteric drugs.
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G protein-coupled receptors (GPCRs): advances in structures, mechanisms and drug discovery
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- Authors
- Vuckovic, Z., Wang, J., Pham, V., Mobbs, J. I., Belousoff, M. J., Bhattarai, A., Burger, W. A., Thompson, G., Yeasmin, M., Nawaratne, V., Leach, K., van der Westhuizen, E. T., Khajehali, E., Liang, Y.-L., Glukhova, A., Wootten, D., Lindsley, C. W., Tobin, A., Sexton, P., Danev, R., Valant, C., Miao, Y., Christopoulos, A., & Thal, D. M.
- Affiliations
- Monash University (AU)
- University of Kansas (US)
- Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University
- Vanderbilt University (US)
- University of Glasgow (GB)
- The University of Tokyo (JP)
- Corresponding author
- Miao, Yinglong
- Christopoulos, Arthur
- Thal, David M
- ORCID
- Venue
- eLife
- Publisher
- eLife Sciences Publications, Ltd
- Published
- 2023-05-30
- Volume
- 12
- ISSN
- 2050-084X (ISSN-L)
- PMC
- PMC10229135
- OpenAlex
- W4378718044
- Cited by
- 34
- References
- 136
- Citations by year
- 2026: 6 · 2025: 13 · 2024: 10 · 2023: 5
- FWCI
- 3.03
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- eLife (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- ENLIGHTEN (Jurnal Bimbingan dan Konseling Islam) (acceptedVersion, cc-by-sa)
- KU ScholarWorks (The University of Kansas) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Wellcome Trust (201529/Z/16/Z)
- National Health and Medical Research Council (1055134, 1150083, 1138448, 1154434, 1155302, 1196951)
- Australian Research Council (DE170100152, DP190102950, IC200100052, 160100075)
- National Institutes of Health (GM132572)
- Takeda Science Foundation (2019 Medical Research Grant)
- Japan Science and Technology Agency (18069571)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Keywords
- M4 muscarinic acetylcholine receptor, allosteric modulation, positive allosteric modulators, acetylcholine, G protein-coupled receptors, receptor conformational dynamics, receptor-G protein complex, cryo-EM structure, molecular dynamics simulation
- MeSH terms
- Acetylcholine, Allosteric Regulation, Allosteric Site, Humans, Ligands, Nicotinic Acids, Pyridazines, Receptors, Muscarinic, Thiophenes, Cryoelectron Microscopy, Receptor, Muscarinic M4
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