i-derived peptide binds the µ-opioid receptor

Pharmacol Rep. 2023 Apr;75(2):465-473. doi: 10.1007/s43440-023-00457-5. Epub 2023 Feb 25.

Abstract

Background: G protein-coupled receptors (GPCRs) transduce external stimuli into the cell by G proteins via an allosteric mechanism. Agonist binding to the receptor stimulates GDP/GTP exchange within the heterotrimeric G protein complex, whereas recent structures of GPCR-G protein complexes revealed that the H5, S1 and S2 domains of Gα are involved in binding the active receptor, earlier studies showed that a short peptide analog derived from the C-terminus (H5) of the G protein transducin (Gt) is sufficient to stabilize rhodopsin in an active form.

Methods: We have used Molecular Dynamics simulations along with biological evaluation by means of radio-ligand binding assay to study the interactions between Gαi-derived peptide (G-peptide) and the µ-opioid receptor (µOR).

Results: Here, we show that a Gαi-derived peptide of 12 amino acids binds the µ-opioid receptor and acts as an allosteric modulator. The Gαi-derived peptide increases µOR affinity for its agonist morphine in a dose-dependent way.

Conclusions: These results indicate that the GPCR-Gα peptide interaction observed so far for only rhodopsin can be extrapolated to µOR. In addition, we show that the C-terminal peptide of the Gαi subunit is sufficient to stabilize the active conformation of the receptor. Our approach opens the possibility to investigate the GPCR-G protein interface with peptide modification.

Keywords: Functional assay; G protein coupling; G protein-coupled receptors; GPCR; Molecular dynamics simulation; µ-opioid receptor.

MeSH terms

  • GTP-Binding Proteins / metabolism
  • Peptides
  • Protein Binding
  • Receptors, G-Protein-Coupled / metabolism
  • Receptors, Opioid* / metabolism
  • Rhodopsin* / chemistry
  • Rhodopsin* / metabolism
  • Transducin / chemistry
  • Transducin / metabolism

Substances

  • Rhodopsin
  • Receptors, Opioid
  • Peptides
  • Receptors, G-Protein-Coupled
  • GTP-Binding Proteins
  • Transducin