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

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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.

Original language	English
Number of pages	11
Journal	Journal of the Royal Society Interface
Volume	10
Issue number	88
DOIs	https://doi.org/10.1098/rsif.2013.0589
Publication status	Published - 6 Nov 2013
Externally published	Yes

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.

Keywords

Calcitonin gene-related peptide
Calcitonin receptor-like receptor
Class B G-protein-coupled receptor
Loop modelling
Site-directed mutagenesis

ASJC Scopus subject areas

Biotechnology
Biophysics
Bioengineering
Biomaterials
Biochemistry
Biomedical Engineering

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10.1098/rsif.2013.0589Licence: CC BY

PublishedFinal published version, 994 KBLicence: CC BY

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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.",

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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.

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

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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