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Receptor Activity-modifying Protein-directed G Protein Signaling Specificity for the Calcitonin Gene-related Peptide Family of Receptors

Weston, Cathryn and Winfield, Ian and Harris, Matthew and Hodgson, Rose and Shah, Archna and Dowell, Simon J and Mobarec, Juan Carlos and Woodlock, David A and Reynolds, Christopher A and Poyner, David R and Watkins, Harriet A and Ladds, Graham (2016) 'Receptor Activity-modifying Protein-directed G Protein Signaling Specificity for the Calcitonin Gene-related Peptide Family of Receptors.' Journal of Biological Chemistry, 291 (42). 21925 - 21944. ISSN 0021-9258

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Abstract

The calcitonin gene-related peptide (CGRP) family of G protein-coupled receptors (GPCRs) is formed through the association of the calcitonin receptor-like receptor (CLR) and one of three receptor activity-modifying proteins (RAMPs). Binding of one of the three peptide ligands, CGRP, adrenomedullin (AM), and intermedin/adrenomedullin 2 (AM2), is well known to result in a Gαs-mediated increase in cAMP. Here we used modified yeast strains that couple receptor activation to cell growth, via chimeric yeast/Gα subunits, and HEK-293 cells to characterize the effect of different RAMP and ligand combinations on this pathway. We not only demonstrate functional couplings to both Gαs and Gαq but also identify a Gαi component to CLR signaling in both yeast and HEK-293 cells, which is absent in HEK-293S cells. We show that the CGRP family of receptors displays both ligand- and RAMP-dependent signaling bias among the Gαs, Gαi, and Gαq/11 pathways. The results are discussed in the context of RAMP interactions probed through molecular modeling and molecular dynamics simulations of the RAMP-GPCR-G protein complexes. This study further highlights the importance of RAMPs to CLR pharmacology and to bias in general, as well as identifying the importance of choosing an appropriate model system for the study of GPCR pharmacology.

Item Type: Article
Uncontrolled Keywords: G protein-coupled receptor (GPCR); molecular dynamics; molecular modeling; signal transduction; yeast; adrenomedullin; adrenomedullin 2; CGRP; RAMP; signal bias
Subjects: Q Science > Q Science (General)
Q Science > QD Chemistry
Divisions: Faculty of Science and Health > Biological Sciences, School of
Depositing User: Christopher Reynolds
Date Deposited: 06 Oct 2016 12:14
Last Modified: 18 Sep 2018 13:15
URI: http://repository.essex.ac.uk/id/eprint/17715

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