Receptor component protein, an endogenous allosteric modulator of family B G protein coupled receptors

Sarah J. Routledge, John Simms, Ashley Clark, Ho Yan Yeung, Mark J. Wigglesworth, Ian M. Dickerson, Philip Kitchen, Graham Ladds, David R. Poyner

Research output: Contribution to journalArticle

Abstract

Receptor component protein (RCP) is a 148 amino acid intracellular peripheral membrane protein, previously identified as promoting the coupling of CGRP to cAMP production at the CGRP receptor, a heterodimer of calcitonin receptor like-receptor (CLR), a family B G protein-coupled receptor (GPCR) and receptor activity modifying protein 1 (RAMP1). We extend these observations to show that it selectively enhances CGRP receptor coupling to Gs but not Gq or pERK activation. At other family B GPCRs, it enhances cAMP production at the calcitonin, corticotrophin releasing factor type 1a and glucagon-like peptide type 2 receptors with their cognate ligands but not at the adrenomedullin type 1 (AM1), gastric inhibitory peptide and glucagon-like peptide type 1 receptors, all expressed in transfected HEK293S cells. However, there is also cell-line variability as RCP did not enhance cAMP production at the endogenous calcitonin receptor in HEK293T cells and it has previously been reported that it is active on the AM1 receptor expressed on NIH3T3 cells. RCP appears to behave as a positive allosteric modulator at coupling a number of family B GPCRs to Gs, albeit in a manner that is regulated by cell-specific factors. It may exert its effects at the interface between the 2nd intracellular loop of the GPCR and Gs, although there is likely to be some overlap between this location and that occupied by the C-terminus of RAMPs if they bind to the GPCRs.
Original languageEnglish
Article number183174
JournalBBA -Biomembranes
Volume1862
Issue number3
Early online date27 Dec 2019
DOIs
Publication statusE-pub ahead of print - 27 Dec 2019

Fingerprint

G-Protein-Coupled Receptors
Calcitonin Gene-Related Peptide Receptors
Modulators
Adrenomedullin
Receptor Activity-Modifying Protein 1
Calcitonin Receptor-Like Protein
Calcitonin Receptors
Proteins
Adrenomedullin Receptors
Corticotropin-Releasing Hormone
Calcitonin
Intracellular Membranes
Peptides
Membrane Proteins
Chemical activation
Ligands
Amino Acids
Stomach
Cell Line

Bibliographical note

© 2019 Published by Elsevier B.V. CC BY-NC-ND.

Funding: Biotechnology and Biological Sciences Research Council [grant number BB/M000176/2]. Rosetrees foundation; international scholarship from the Cambridge Trust. Aston University 50th Anniversary Prize Fellowship. EPSRC on grant EP/P020232/1 as part of HPC Midlands+ consortium.

Keywords

  • Adrenomedullin; Accessory protein
  • GPCR signalling; CGRP
  • G protein coupling
  • Allosteric modulator

Cite this

Routledge, Sarah J. ; Simms, John ; Clark, Ashley ; Yeung, Ho Yan ; Wigglesworth, Mark J. ; Dickerson, Ian M. ; Kitchen, Philip ; Ladds, Graham ; Poyner, David R. / Receptor component protein, an endogenous allosteric modulator of family B G protein coupled receptors. In: BBA -Biomembranes. 2020 ; Vol. 1862, No. 3.
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Receptor component protein, an endogenous allosteric modulator of family B G protein coupled receptors. / Routledge, Sarah J.; Simms, John; Clark, Ashley; Yeung, Ho Yan; Wigglesworth, Mark J.; Dickerson, Ian M.; Kitchen, Philip; Ladds, Graham; Poyner, David R.

In: BBA -Biomembranes, Vol. 1862, No. 3, 183174, 01.03.2020.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Receptor component protein, an endogenous allosteric modulator of family B G protein coupled receptors

AU - Routledge, Sarah J.

AU - Simms, John

AU - Clark, Ashley

AU - Yeung, Ho Yan

AU - Wigglesworth, Mark J.

AU - Dickerson, Ian M.

AU - Kitchen, Philip

AU - Ladds, Graham

AU - Poyner, David R.

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PY - 2019/12/27

Y1 - 2019/12/27

N2 - Receptor component protein (RCP) is a 148 amino acid intracellular peripheral membrane protein, previously identified as promoting the coupling of CGRP to cAMP production at the CGRP receptor, a heterodimer of calcitonin receptor like-receptor (CLR), a family B G protein-coupled receptor (GPCR) and receptor activity modifying protein 1 (RAMP1). We extend these observations to show that it selectively enhances CGRP receptor coupling to Gs but not Gq or pERK activation. At other family B GPCRs, it enhances cAMP production at the calcitonin, corticotrophin releasing factor type 1a and glucagon-like peptide type 2 receptors with their cognate ligands but not at the adrenomedullin type 1 (AM1), gastric inhibitory peptide and glucagon-like peptide type 1 receptors, all expressed in transfected HEK293S cells. However, there is also cell-line variability as RCP did not enhance cAMP production at the endogenous calcitonin receptor in HEK293T cells and it has previously been reported that it is active on the AM1 receptor expressed on NIH3T3 cells. RCP appears to behave as a positive allosteric modulator at coupling a number of family B GPCRs to Gs, albeit in a manner that is regulated by cell-specific factors. It may exert its effects at the interface between the 2nd intracellular loop of the GPCR and Gs, although there is likely to be some overlap between this location and that occupied by the C-terminus of RAMPs if they bind to the GPCRs.

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KW - Adrenomedullin; Accessory protein

KW - GPCR signalling; CGRP

KW - G protein coupling

KW - Allosteric modulator

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