Propagation of conformational changes during μ-opioid receptor activation.

TitlePropagation of conformational changes during μ-opioid receptor activation.
Publication TypeJournal Article
Year of Publication2015
AuthorsSounier, R., Mas C., Steyaert J., Laeremans T., Manglik A., Huang W., Kobilka B. K., Déméné H., and Granier S.
JournalNature
Volume524
Issue7565
Pagination375-8
Date Published2015 Aug 20
ISSN1476-4687
KeywordsAllosteric Regulation, Animals, Binding Sites, Heterotrimeric GTP-Binding Proteins, Lysine, Mice, Models, Molecular, Morphinans, Nuclear Magnetic Resonance, Biomolecular, Protein Binding, Protein Conformation, Pyrroles, Receptors, Adrenergic, beta-2, Receptors, Opioid, mu, Single-Chain Antibodies, Structure-Activity Relationship, Substrate Specificity
Abstract

µ-Opioid receptors (µORs) are G-protein-coupled receptors that are activated by a structurally diverse spectrum of natural and synthetic agonists including endogenous endorphin peptides, morphine and methadone. The recent structures of the μOR in inactive and agonist-induced active states (Huang et al., ref. 2) provide snapshots of the receptor at the beginning and end of a signalling event, but little is known about the dynamic sequence of events that span these two states. Here we use solution-state NMR to examine the process of μOR activation using a purified receptor (mouse sequence) preparation in an amphiphile membrane-like environment. We obtain spectra of the μOR in the absence of ligand, and in the presence of the high-affinity agonist BU72 alone, or with BU72 and a G protein mimetic nanobody. Our results show that conformational changes in transmembrane segments 5 and 6 (TM5 and TM6), which are required for the full engagement of a G protein, are almost completely dependent on the presence of both the agonist and the G protein mimetic nanobody, revealing a weak allosteric coupling between the agonist-binding pocket and the G-protein-coupling interface (TM5 and TM6), similar to that observed for the β2-adrenergic receptor. Unexpectedly, in the presence of agonist alone, we find larger spectral changes involving intracellular loop 1 and helix 8 compared to changes in TM5 and TM6. These results suggest that one or both of these domains may play a role in the initial interaction with the G protein, and that TM5 and TM6 are only engaged later in the process of complex formation. The initial interactions between the G protein and intracellular loop 1 and/or helix 8 may be involved in G-protein coupling specificity, as has been suggested for other family A G-protein-coupled receptors.

DOI10.1038/nature14680
Alternate JournalNature
PubMed ID26245377
PubMed Central IDPMC4820006
Grant ListR37 DA036246 / DA / NIDA NIH HHS / United States
T32 GM008294 / GM / NIGMS NIH HHS / United States
DA036246 / DA / NIDA NIH HHS / United States
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