An allosteric role for receptor activity-modifying proteins in defining GPCR pharmacology

Joseph J Gingell, John Simms, James Barwell, David R Poyner, Harriet A Watkins, Augen A Pioszak, Patrick M Sexton, Debbie L Hay

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

G protein-coupled receptors are allosteric proteins that control transmission of external signals to regulate cellular response. Although agonist binding promotes canonical G protein signalling transmitted through conformational changes, G protein-coupled receptors also interact with other proteins. These include other G protein-coupled receptors, other receptors and channels, regulatory proteins and receptor-modifying proteins, notably receptor activity-modifying proteins (RAMPs). RAMPs have at least 11 G protein-coupled receptor partners, including many class B G protein-coupled receptors. Prototypic is the calcitonin receptor, with altered ligand specificity when co-expressed with RAMPs. To gain molecular insight into the consequences of this protein-protein interaction, we combined molecular modelling with mutagenesis of the calcitonin receptor extracellular domain, assessed in ligand binding and functional assays. Although some calcitonin receptor residues are universally important for peptide interactions (calcitonin, amylin and calcitonin gene-related peptide) in calcitonin receptor alone or with receptor activity-modifying protein, others have RAMP-dependent effects, whereby mutations decreased amylin/calcitonin gene-related peptide potency substantially only when RAMP was present. Remarkably, the key residues were completely conserved between calcitonin receptor and AMY receptors, and between subtypes of AMY receptor that have different ligand preferences. Mutations at the interface between calcitonin receptor and RAMP affected ligand pharmacology in a RAMP-dependent manner, suggesting that RAMP may allosterically influence the calcitonin receptor conformation. Supporting this, molecular dynamics simulations suggested that the calcitonin receptor extracellular N-terminal domain is more flexible in the presence of receptor activity-modifying protein 1. Thus, RAMPs may act in an allosteric manner to generate a spectrum of unique calcitonin receptor conformational states, explaining the pharmacological preferences of calcitonin receptor-RAMP complexes. This provides novel insight into our understanding of G protein-coupled receptor-protein interaction that is likely broadly applicable for this receptor class.

Original languageEnglish
Article number16012
Pages (from-to)1-14
Number of pages15
JournalCell Discovery
Volume2
DOIs
Publication statusPublished - 2016
Externally publishedYes

Fingerprint

Receptor Activity-Modifying Proteins
Calcitonin Receptors
Pharmacology
G-Protein-Coupled Receptors
Proteins
Islet Amyloid Polypeptide
Ligands
Calcitonin Gene-Related Peptide
Receptor Activity-Modifying Protein 1
Mutagenesis
Mutation
Molecular modeling
Calcitonin
Molecular Dynamics Simulation
GTP-Binding Proteins

Keywords

  • Amylin
  • accessory protein
  • CGRP
  • G protein-coupled receptor
  • RAMP Cell Discovery

Cite this

J Gingell, J., Simms, J., Barwell, J., Poyner, D. R., Watkins, H. A., Pioszak, A. A., ... Hay, D. L. (2016). An allosteric role for receptor activity-modifying proteins in defining GPCR pharmacology. Cell Discovery, 2, 1-14. [16012]. https://doi.org/10.1038/celldisc.2016.12

An allosteric role for receptor activity-modifying proteins in defining GPCR pharmacology. / J Gingell, Joseph; Simms, John; Barwell, James; Poyner, David R; Watkins, Harriet A; Pioszak, Augen A; Sexton, Patrick M; Hay, Debbie L.

In: Cell Discovery, Vol. 2, 16012, 2016, p. 1-14.

Research output: Contribution to journalArticle

J Gingell, J, Simms, J, Barwell, J, Poyner, DR, Watkins, HA, Pioszak, AA, Sexton, PM & Hay, DL 2016, 'An allosteric role for receptor activity-modifying proteins in defining GPCR pharmacology' Cell Discovery, vol. 2, 16012, pp. 1-14. https://doi.org/10.1038/celldisc.2016.12
J Gingell J, Simms J, Barwell J, Poyner DR, Watkins HA, Pioszak AA et al. An allosteric role for receptor activity-modifying proteins in defining GPCR pharmacology. Cell Discovery. 2016;2:1-14. 16012. https://doi.org/10.1038/celldisc.2016.12
J Gingell, Joseph ; Simms, John ; Barwell, James ; Poyner, David R ; Watkins, Harriet A ; Pioszak, Augen A ; Sexton, Patrick M ; Hay, Debbie L. / An allosteric role for receptor activity-modifying proteins in defining GPCR pharmacology. In: Cell Discovery. 2016 ; Vol. 2. pp. 1-14.
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