Modulation of glucagon receptor pharmacology by Receptor Activity-modifying Protein-2 (RAMP2)

Cathryn Weston*, Jing Lu, Naichang Li, Kerry Barkan, Gareth O. Richards, David J. Roberts, Timothy M. Skerry, David Poyner, Meenakshi Pardamwar, Christopher A. Reynolds, Simon J. Dowell, Gary B. Willars, Graham Ladds

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

The glucagon and glucagon-like peptide-1 (GLP-1) receptors play important, opposing roles in regulating blood glucose levels. Consequently, these receptors have been identified as targets for novel diabetes treatments. However, drugs acting at the GLP-1 receptor, whilst having clinical efficacy, have been associated with severe adverse side-effects and targeting of the glucagon receptor has yet to be successful. Here we use a combination of yeast reporter assays and mammalian systems, to provide a more complete understanding of glucagon receptor signaling considering the effect of multiple ligands, association with the receptor-interacting protein, receptor activity modifying protein-2 (RAMP2) and individual G protein α-subunits. We demonstrate that RAMP2 alters both ligand selectivity and G protein preference of the glucagon receptor. Importantly, we also uncover novel cross-reactivity of therapeutically used GLP-1 receptor ligands at the glucagon receptor that is abolished by RAMP2 interaction. This study reveals the glucagon receptor as a previously unidentified target for GLP-1 receptor agonists and highlights a role for RAMP2 in regulating its pharmacology. Such previously unrecognized functions of RAMPs highlight the need to consider all receptor-interacting proteins in future drug development.

Original languageEnglish
Pages (from-to)23009-23022
Number of pages14
JournalJournal of Biological Chemistry
Volume290
Issue number38
Early online date21 Jul 2015
DOIs
Publication statusPublished - 18 Sep 2015

Fingerprint

Receptor Activity-Modifying Protein 2
Glucagon Receptors
Modulation
Pharmacology
Receptor-Interacting Protein Serine-Threonine Kinases
Ligands
GTP-Binding Proteins
Protein Subunits
Medical problems
Glucagon
Pharmaceutical Preparations
Yeast
Blood Glucose
Assays
Yeasts
Association reactions
Glucagon-Like Peptide-1 Receptor

Bibliographical note

© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.
Final version free via Creative Commons CC-BY licence.

Funding: Warwick Impact Fund; BBSRC (BB/G01227X/1; BB/F008392/1, BB/M007529/1 and BB/M000176/1); Warwick Research Development Fund (RD13301) and the Birmingham Science City Research Alliance

Keywords

  • G protein-coupled receptor
  • glucagon
  • pharmacology
  • signal transduction
  • type 2 diabetes
  • glucagon receptor
  • glucagon-like peptide-1
  • receptor activity modifying proteins
  • RAMPs
  • GPCR
  • signal bias

Cite this

Weston, C., Lu, J., Li, N., Barkan, K., Richards, G. O., Roberts, D. J., ... Ladds, G. (2015). Modulation of glucagon receptor pharmacology by Receptor Activity-modifying Protein-2 (RAMP2). Journal of Biological Chemistry, 290(38), 23009-23022. https://doi.org/10.1074/jbc.M114.624601
Weston, Cathryn ; Lu, Jing ; Li, Naichang ; Barkan, Kerry ; Richards, Gareth O. ; Roberts, David J. ; Skerry, Timothy M. ; Poyner, David ; Pardamwar, Meenakshi ; Reynolds, Christopher A. ; Dowell, Simon J. ; Willars, Gary B. ; Ladds, Graham. / Modulation of glucagon receptor pharmacology by Receptor Activity-modifying Protein-2 (RAMP2). In: Journal of Biological Chemistry. 2015 ; Vol. 290, No. 38. pp. 23009-23022.
@article{056bc16f649245f7ab4b0510582328e7,
title = "Modulation of glucagon receptor pharmacology by Receptor Activity-modifying Protein-2 (RAMP2)",
abstract = "The glucagon and glucagon-like peptide-1 (GLP-1) receptors play important, opposing roles in regulating blood glucose levels. Consequently, these receptors have been identified as targets for novel diabetes treatments. However, drugs acting at the GLP-1 receptor, whilst having clinical efficacy, have been associated with severe adverse side-effects and targeting of the glucagon receptor has yet to be successful. Here we use a combination of yeast reporter assays and mammalian systems, to provide a more complete understanding of glucagon receptor signaling considering the effect of multiple ligands, association with the receptor-interacting protein, receptor activity modifying protein-2 (RAMP2) and individual G protein α-subunits. We demonstrate that RAMP2 alters both ligand selectivity and G protein preference of the glucagon receptor. Importantly, we also uncover novel cross-reactivity of therapeutically used GLP-1 receptor ligands at the glucagon receptor that is abolished by RAMP2 interaction. This study reveals the glucagon receptor as a previously unidentified target for GLP-1 receptor agonists and highlights a role for RAMP2 in regulating its pharmacology. Such previously unrecognized functions of RAMPs highlight the need to consider all receptor-interacting proteins in future drug development.",
keywords = "G protein-coupled receptor, glucagon, pharmacology, signal transduction, type 2 diabetes, glucagon receptor, glucagon-like peptide-1, receptor activity modifying proteins, RAMPs, GPCR, signal bias",
author = "Cathryn Weston and Jing Lu and Naichang Li and Kerry Barkan and Richards, {Gareth O.} and Roberts, {David J.} and Skerry, {Timothy M.} and David Poyner and Meenakshi Pardamwar and Reynolds, {Christopher A.} and Dowell, {Simon J.} and Willars, {Gary B.} and Graham Ladds",
note = "{\circledC} 2015 by The American Society for Biochemistry and Molecular Biology, Inc. Final version free via Creative Commons CC-BY licence. Funding: Warwick Impact Fund; BBSRC (BB/G01227X/1; BB/F008392/1, BB/M007529/1 and BB/M000176/1); Warwick Research Development Fund (RD13301) and the Birmingham Science City Research Alliance",
year = "2015",
month = "9",
day = "18",
doi = "10.1074/jbc.M114.624601",
language = "English",
volume = "290",
pages = "23009--23022",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "38",

}

Weston, C, Lu, J, Li, N, Barkan, K, Richards, GO, Roberts, DJ, Skerry, TM, Poyner, D, Pardamwar, M, Reynolds, CA, Dowell, SJ, Willars, GB & Ladds, G 2015, 'Modulation of glucagon receptor pharmacology by Receptor Activity-modifying Protein-2 (RAMP2)', Journal of Biological Chemistry, vol. 290, no. 38, pp. 23009-23022. https://doi.org/10.1074/jbc.M114.624601

Modulation of glucagon receptor pharmacology by Receptor Activity-modifying Protein-2 (RAMP2). / Weston, Cathryn; Lu, Jing; Li, Naichang; Barkan, Kerry; Richards, Gareth O.; Roberts, David J.; Skerry, Timothy M.; Poyner, David; Pardamwar, Meenakshi; Reynolds, Christopher A.; Dowell, Simon J.; Willars, Gary B.; Ladds, Graham.

In: Journal of Biological Chemistry, Vol. 290, No. 38, 18.09.2015, p. 23009-23022.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Modulation of glucagon receptor pharmacology by Receptor Activity-modifying Protein-2 (RAMP2)

AU - Weston, Cathryn

AU - Lu, Jing

AU - Li, Naichang

AU - Barkan, Kerry

AU - Richards, Gareth O.

AU - Roberts, David J.

AU - Skerry, Timothy M.

AU - Poyner, David

AU - Pardamwar, Meenakshi

AU - Reynolds, Christopher A.

AU - Dowell, Simon J.

AU - Willars, Gary B.

AU - Ladds, Graham

N1 - © 2015 by The American Society for Biochemistry and Molecular Biology, Inc. Final version free via Creative Commons CC-BY licence. Funding: Warwick Impact Fund; BBSRC (BB/G01227X/1; BB/F008392/1, BB/M007529/1 and BB/M000176/1); Warwick Research Development Fund (RD13301) and the Birmingham Science City Research Alliance

PY - 2015/9/18

Y1 - 2015/9/18

N2 - The glucagon and glucagon-like peptide-1 (GLP-1) receptors play important, opposing roles in regulating blood glucose levels. Consequently, these receptors have been identified as targets for novel diabetes treatments. However, drugs acting at the GLP-1 receptor, whilst having clinical efficacy, have been associated with severe adverse side-effects and targeting of the glucagon receptor has yet to be successful. Here we use a combination of yeast reporter assays and mammalian systems, to provide a more complete understanding of glucagon receptor signaling considering the effect of multiple ligands, association with the receptor-interacting protein, receptor activity modifying protein-2 (RAMP2) and individual G protein α-subunits. We demonstrate that RAMP2 alters both ligand selectivity and G protein preference of the glucagon receptor. Importantly, we also uncover novel cross-reactivity of therapeutically used GLP-1 receptor ligands at the glucagon receptor that is abolished by RAMP2 interaction. This study reveals the glucagon receptor as a previously unidentified target for GLP-1 receptor agonists and highlights a role for RAMP2 in regulating its pharmacology. Such previously unrecognized functions of RAMPs highlight the need to consider all receptor-interacting proteins in future drug development.

AB - The glucagon and glucagon-like peptide-1 (GLP-1) receptors play important, opposing roles in regulating blood glucose levels. Consequently, these receptors have been identified as targets for novel diabetes treatments. However, drugs acting at the GLP-1 receptor, whilst having clinical efficacy, have been associated with severe adverse side-effects and targeting of the glucagon receptor has yet to be successful. Here we use a combination of yeast reporter assays and mammalian systems, to provide a more complete understanding of glucagon receptor signaling considering the effect of multiple ligands, association with the receptor-interacting protein, receptor activity modifying protein-2 (RAMP2) and individual G protein α-subunits. We demonstrate that RAMP2 alters both ligand selectivity and G protein preference of the glucagon receptor. Importantly, we also uncover novel cross-reactivity of therapeutically used GLP-1 receptor ligands at the glucagon receptor that is abolished by RAMP2 interaction. This study reveals the glucagon receptor as a previously unidentified target for GLP-1 receptor agonists and highlights a role for RAMP2 in regulating its pharmacology. Such previously unrecognized functions of RAMPs highlight the need to consider all receptor-interacting proteins in future drug development.

KW - G protein-coupled receptor

KW - glucagon

KW - pharmacology

KW - signal transduction

KW - type 2 diabetes

KW - glucagon receptor

KW - glucagon-like peptide-1

KW - receptor activity modifying proteins

KW - RAMPs

KW - GPCR

KW - signal bias

UR - http://www.scopus.com/inward/record.url?scp=84942860294&partnerID=8YFLogxK

U2 - 10.1074/jbc.M114.624601

DO - 10.1074/jbc.M114.624601

M3 - Article

C2 - 26198634

VL - 290

SP - 23009

EP - 23022

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 38

ER -