title: Receptor Activity-modifying Protein-directed G Protein Signaling Specificity for the Calcitonin Gene-related Peptide Family of Receptors.
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 Gαsand Gαqbut also identify a Gαicomponent 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/11pathways. 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.
authors: From the Division of Biomedical Cell Biology, Warwick Medical School, University of Warwick, Coventry, CV4 7AL, United Kingdom.
authors: Cathryn
authors: C
authors: Weston
authors: From the Division of Biomedical Cell Biology, Warwick Medical School, University of Warwick, Coventry, CV4 7AL, United Kingdom, the Department of Pharmacology, University of Cambridge, Cambridge, CB2 1PD, United Kingdom.
authors: Ian
authors: I
authors: Winfield
authors: the Department of Pharmacology, University of Cambridge, Cambridge, CB2 1PD, United Kingdom.
authors: Matthew
authors: M
authors: Harris
authors: From the Division of Biomedical Cell Biology, Warwick Medical School, University of Warwick, Coventry, CV4 7AL, United Kingdom.
authors: Rose
authors: R
authors: Hodgson
authors: From the Division of Biomedical Cell Biology, Warwick Medical School, University of Warwick, Coventry, CV4 7AL, United Kingdom.
authors: Archna
authors: A
authors: Shah
authors: the Department of Platform Technology and Science, GlaxoSmithkline, Hertfordshire, SG1 2NY, United Kingdom.
authors: Simon J
authors: SJ
authors: Dowell
authors: the School of Biological Sciences, University of Essex, Wivenhoe Park, Colchester, Essex, CO4 3SQ, United Kingdom.
authors: Juan Carlos
authors: JC
authors: Mobarec
authors: the School of Biological Sciences, University of Essex, Wivenhoe Park, Colchester, Essex, CO4 3SQ, United Kingdom.
authors: David A
authors: DA
authors: Woodlock
authors: the School of Biological Sciences, University of Essex, Wivenhoe Park, Colchester, Essex, CO4 3SQ, United Kingdom.
authors: Christopher A
authors: CA
authors: Reynolds
authors: the School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, B4 7ET, United Kingdom, and.
authors: David R
authors: DR
authors: Poyner
authors: the School of Biological Sciences and Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland 1010, New Zealand.
authors: Harriet A
authors: HA
authors: Watkins
authors: the Department of Pharmacology, University of Cambridge, Cambridge, CB2 1PD, United Kingdom, grl30@cam.ac.uk.
authors: Graham
authors: G
authors: Ladds
mesh_terms: D053607:Adrenomedullin / Q000235:genetics / Q000378:metabolism*; D000242:Cyclic AMP / Q000235:genetics / Q000378:metabolism*; D044385:GTP-Binding Protein alpha Subunits / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D018015:Receptors, Calcitonin Gene-Related Peptide / Q000235:genetics / Q000378:metabolism*; D015290:Second Messenger Systems / Q000502:physiology*
chemical_list: C429530:DTL protein, human; D044385:GTP-Binding Protein alpha Subunits; D009687:Nuclear Proteins; D018015:Receptors, Calcitonin Gene-Related Peptide; D053607:Adrenomedullin; D000242:Cyclic AMP
keywords: CGRP; G protein-coupled receptor (GPCR); RAMP; adrenomedullin; adrenomedullin 2; molecular dynamics; molecular modeling; signal bias; signal transduction; yeast