The role of ECL2 in CGRP receptor activation: a combined modelling and experimental approach

Michael J. Woolley; Harriet A. Watkins; Bruck Taddese; Z. Gamze Karakullukcu; James Barwell; Kevin J. Smith; Debbie L. Hay; David R. Poyner; Christopher A. Reynolds; Alex C. Conner

doi:10.1098/rsif.2013.0589

The role of ECL2 in CGRP receptor activation: a combined modelling and experimental approach

Michael J. Woolley, Harriet A. Watkins, Bruck Taddese, Z. Gamze Karakullukcu, James Barwell, Kevin J. Smith, Debbie L. Hay, David R. Poyner^*, Christopher A. Reynolds, Alex C. Conner

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

The calcitonin gene-related peptide (CGRP) receptor is a complex of a cal-citonin receptor-like receptor (CLR), which is a family B G-protein-coupled receptor (GPCR) and receptor activity modifying protein 1. The role of the second extracellular loop (ECL2) of CLR in binding CGRP and coupling to Gs was investigated using a combination of mutagenesis and modelling. An alanine scan of residues 271-294 of CLR showed that the ability of CGRP to produce cAMP was impaired by point mutations at 13 residues; most of these also impaired the response to adrenomedullin (AM). These data were used to select probable ECL2-modelled conformations that are involved in agonist binding, allowing the identification of the likely contacts between the peptide and receptor. The implications of the most likely structures for receptor activation are discussed. © 2013 The Authors.

Original language	English
Article number	20130589
Number of pages	11
Journal	Journal of the Royal Society Interface
Volume	10
Issue number	88
Early online date	18 Sept 2013
DOIs	https://doi.org/10.1098/rsif.2013.0589
Publication status	Published - Nov 2013

Bibliographical note

© 2013 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses by/3.0/, which permits unrestricted use, provided the original author and source are credited.

Funding: Wellcome Trust [091496]; New Zealand Heart Foundation; MRC [G1001812]

Keywords

calcitonin gene-related peptide
calcitonin receptor-like receptor
class B G-protein-coupled receptor
loop modelling
site-directed mutagenesis

Access to Document

10.1098/rsif.2013.0589

Role of ECL2 in CGRP receptor activation
© 2013 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses by/3.0/, which permits unrestricted use, provided the original author and source are credited.
Final published version, 1.09 MBLicence: CC BY 3.0

http://rsif.royalsocietypublishing.org/content/10/88/20130589

Cite this

@article{4689681d0ac34e27a23961ca14589a3f,

title = "The role of ECL2 in CGRP receptor activation: a combined modelling and experimental approach",

abstract = "The calcitonin gene-related peptide (CGRP) receptor is a complex of a cal-citonin receptor-like receptor (CLR), which is a family B G-protein-coupled receptor (GPCR) and receptor activity modifying protein 1. The role of the second extracellular loop (ECL2) of CLR in binding CGRP and coupling to Gs was investigated using a combination of mutagenesis and modelling. An alanine scan of residues 271-294 of CLR showed that the ability of CGRP to produce cAMP was impaired by point mutations at 13 residues; most of these also impaired the response to adrenomedullin (AM). These data were used to select probable ECL2-modelled conformations that are involved in agonist binding, allowing the identification of the likely contacts between the peptide and receptor. The implications of the most likely structures for receptor activation are discussed. {\textcopyright} 2013 The Authors.",

keywords = "calcitonin gene-related peptide, calcitonin receptor-like receptor, class B G-protein-coupled receptor, loop modelling, site-directed mutagenesis",

author = "Woolley, {Michael J.} and Watkins, {Harriet A.} and Bruck Taddese and Karakullukcu, {Z. Gamze} and James Barwell and Smith, {Kevin J.} and Hay, {Debbie L.} and Poyner, {David R.} and Reynolds, {Christopher A.} and Conner, {Alex C.}",

note = "{\textcopyright} 2013 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses by/3.0/, which permits unrestricted use, provided the original author and source are credited. Funding: Wellcome Trust [091496]; New Zealand Heart Foundation; MRC [G1001812]",

year = "2013",

month = nov,

doi = "10.1098/rsif.2013.0589",

language = "English",

volume = "10",

journal = "Journal of the Royal Society Interface",

issn = "1742-5662",

publisher = "Royal Society of London",

number = "88",

}

TY - JOUR

T1 - The role of ECL2 in CGRP receptor activation

T2 - a combined modelling and experimental approach

AU - Woolley, Michael J.

AU - Watkins, Harriet A.

AU - Taddese, Bruck

AU - Karakullukcu, Z. Gamze

AU - Barwell, James

AU - Smith, Kevin J.

AU - Hay, Debbie L.

AU - Poyner, David R.

AU - Reynolds, Christopher A.

AU - Conner, Alex C.

N1 - © 2013 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses by/3.0/, which permits unrestricted use, provided the original author and source are credited. Funding: Wellcome Trust [091496]; New Zealand Heart Foundation; MRC [G1001812]

PY - 2013/11

Y1 - 2013/11

N2 - The calcitonin gene-related peptide (CGRP) receptor is a complex of a cal-citonin receptor-like receptor (CLR), which is a family B G-protein-coupled receptor (GPCR) and receptor activity modifying protein 1. The role of the second extracellular loop (ECL2) of CLR in binding CGRP and coupling to Gs was investigated using a combination of mutagenesis and modelling. An alanine scan of residues 271-294 of CLR showed that the ability of CGRP to produce cAMP was impaired by point mutations at 13 residues; most of these also impaired the response to adrenomedullin (AM). These data were used to select probable ECL2-modelled conformations that are involved in agonist binding, allowing the identification of the likely contacts between the peptide and receptor. The implications of the most likely structures for receptor activation are discussed. © 2013 The Authors.

AB - The calcitonin gene-related peptide (CGRP) receptor is a complex of a cal-citonin receptor-like receptor (CLR), which is a family B G-protein-coupled receptor (GPCR) and receptor activity modifying protein 1. The role of the second extracellular loop (ECL2) of CLR in binding CGRP and coupling to Gs was investigated using a combination of mutagenesis and modelling. An alanine scan of residues 271-294 of CLR showed that the ability of CGRP to produce cAMP was impaired by point mutations at 13 residues; most of these also impaired the response to adrenomedullin (AM). These data were used to select probable ECL2-modelled conformations that are involved in agonist binding, allowing the identification of the likely contacts between the peptide and receptor. The implications of the most likely structures for receptor activation are discussed. © 2013 The Authors.

KW - calcitonin gene-related peptide

KW - calcitonin receptor-like receptor

KW - class B G-protein-coupled receptor

KW - loop modelling

KW - site-directed mutagenesis

UR - http://www.scopus.com/inward/record.url?scp=84884886111&partnerID=8YFLogxK

U2 - 10.1098/rsif.2013.0589

DO - 10.1098/rsif.2013.0589

M3 - Article

C2 - 24047872

SN - 1742-5662

VL - 10

JO - Journal of the Royal Society Interface

JF - Journal of the Royal Society Interface

IS - 88

M1 - 20130589

ER -

The role of ECL2 in CGRP receptor activation: a combined modelling and experimental approach

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

Fingerprint

Cite this