Receptor activity-modifying proteins 2 and 3 generate adrenomedullin receptor subtypes with distinct molecular properties

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Authors

  • Harriet A. Watkins
  • Madhuri Chakravarthy
  • Rekhati S. Abhayawardana
  • Joseph J. Gingell
  • Michael Garelja
  • Meenakshi Pardamwar
  • James M.W.R. McElhinney
  • Alex Lathbridge
  • Arran Constantine
  • Paul W.R. Harris
  • Tsz-Ying Yuen
  • Margaret A. Brimble
  • James Barwell
  • David R. PoynerORCiD: http://orcid.org/0000-0003-1590-112X
  • Michael J. Woolley
  • Alex C. Conner
  • Augen A. Pioszak
  • Christopher A. Reynolds
  • Debbie L. Hay

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Abstract

Adrenomedullin (AM) is a peptide hormone with numerous effects in the vascular systems. AM signals through the AM1 and AM2 receptors formed by the obligate heterodimerization of a G protein-coupled receptor, the calcitonin receptor-like receptor (CLR), and receptor activity-modifying proteins (RAMP) 2 and 3, respectively. These different CLR-RAMP interactions yield discrete receptor pharmacology and physiological effects. The effective design of therapeutics that target the individual AM receptors is dependent on understanding the molecular details of the effects of RAMPs on CLR. To understand the role of RAMPs 2 and 3 on the activation and conformation of the CLR subunit of AM receptors we mutated 68 individual amino acids in the juxtamembrane region of CLR, a key region for activation of AM receptors and determined the effects on cAMP signalling. Sixteen CLR mutations had differential effects between the AM1 and AM2 receptors. Accompanying this, independent molecular modelling of the full-length AM-bound AM1 and AM2 receptors predicted differences in the binding pocket, and differences in the electrostatic potential of the two AM receptors. Druggability analysis indicated unique features that could be used to develop selective small molecule ligands for each receptor. The interaction of RAMP2 or RAMP3 with CLR induces conformational variation in the juxtamembrane region, yielding distinct binding pockets, probably via an allosteric mechanism. These subtype-specific differences have implications for the design of therapeutics aimed at specific AM receptors and for understanding the mechanisms by which accessory proteins affect G protein-coupled receptor function.

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Details

Original languageEnglish
Pages (from-to)11657-11675
Number of pages19
JournalJournal of Biological Chemistry
Volume291
Issue number22
Early online date24 Mar 2016
DOIs
Publication statusPublished - 27 May 2016

Bibliographic note

Final version free via Creative Commons CC-BY license. Funding: BBSRC (BB/M006883/1; BB/M007529/1 and BB/M000176/1) Supplemental material: http://www.jbc.org/content/suppl/2016/04/13/M115.688218.DC1.htm

    Keywords

  • allosteric regulation, cardiovascular disease, conformational change, G protein-coupled receptor, GPCR, molecular modeling, RAMP, adrenomedullin, extracellular loops, receptor activity-modifying protein

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