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 calcitonin 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.
authors.affiliation: Warwick Medical School, University of Warwick, Coventry CV4 7AL, UK.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Woolley
authors.forename: Harriet A
authors.initials: HA
authors.lastname: Watkins
authors.forename: Bruck
authors.initials: B
authors.lastname: Taddese
authors.forename: Z Gamze
authors.initials: ZG
authors.lastname: Karakullukcu
authors.forename: James
authors.initials: J
authors.lastname: Barwell
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Smith
authors.forename: Debbie L
authors.initials: DL
authors.lastname: Hay
authors.forename: David R
authors.initials: DR
authors.lastname: Poyner
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Reynolds
authors.forename: Alex C
authors.initials: AC
authors.lastname: Conner
mesh_terms: D053607:Adrenomedullin / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D058286:Calcitonin Receptor-Like Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D018015:Receptors, Calcitonin Gene-Related Peptide / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: C545578:CALCRL protein, human; D058286:Calcitonin Receptor-Like Protein; D018015:Receptors, Calcitonin Gene-Related Peptide; D053607:Adrenomedullin; D000242:Cyclic AMP
keywords: calcitonin gene-related peptide; calcitonin receptor-like receptor; class B G-protein-coupled receptor; loop modelling; site-directed mutagenesis
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