The second intracellular loop of the calcitonin gene-related peptide receptor provides molecular determinants for signal transduction and cell surface expression


The calcitonin gene-related peptide (CGRP) receptor is a heterodimer of a family B G-protein-coupled receptor, calcitonin receptor-like receptor (CLR), and the accessory protein receptor activity modifying protein 1. It couples to Gs, but it is not known which intracellular loops mediate this. We have identified the boundaries of this loop based on the relative position and length of the juxtamembrane transmembrane regions 3 and 4. The loop has been analyzed by systematic mutagenesis of all residues to alanine, measuring cAMP accumulation, CGRP affinity, and receptor expression. Unlike rhodopsin, ICL2 of the CGRP receptor plays a part in the conformational switch after agonist interaction. His-216 and Lys-227 were essential for a functional CGRP-induced cAMP response. The effect of (H216A)CLR is due to a disruption to the cell surface transport or surface stability of the mutant receptor. In contrast, (K227A)CLR had wild-type expression and agonist affinity, suggesting a direct disruption to the downstream signal transduction mechanism of the CGRP receptor. Modeling suggests that the loop undergoes a significant shift in position during receptor activation, exposing a potential G-protein binding pocket. Lys-227 changes position to point into the pocket, potentially allowing it to interact with bound G-proteins. His-216 occupies a position similar to that of Tyr-136 in bovine rhodopsin, part of the DRY motif of the latter receptor. This is the first comprehensive analysis of an entire intracellular loop within the calcitonin family of G-protein-coupled receptor. These data help to define the structural and functional characteristics of the CGRP-receptor and of family B G-protein-coupled receptors in general. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

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Divisions: College of Health & Life Sciences > School of Biosciences
College of Health & Life Sciences > Aston Pharmacy School
College of Health & Life Sciences > Chronic and Communicable Conditions
College of Health & Life Sciences
Additional Information: © The American Society for Biochemistry and Molecular Biology, Inc.
Uncontrolled Keywords: alanine,amino acid sequence,amino acid substitution,animals,COS,cells,calcitonin,gene-related peptide,cell membrane,cercopithecus,aethiops,cyclic,AMP,DNA,primers,gene expression,regulation,humans kinetics models,molecular sequence data mutagenesis,site-directed protein structure,secondary receptors,calcitonin gene-related peptide,receptors,G-protein-coupled signal transduction,Biochemistry
Publication ISSN: 1083-351X
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Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
http://www.jbc. ... tent/281/3/1644 (Publisher URL)
PURE Output Type: Article
Published Date: 2006-01-20
Published Online Date: 2005-11-17
Authors: Conner, Alex C.
Simms, John (ORCID Profile 0000-0002-4675-0902)
Howitt, Stephen G.
Wheatley, Mark
Poyner, David R. (ORCID Profile 0000-0003-1590-112X)



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