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dc.contributor.authorWeston, Cathryn
dc.contributor.authorWinfield, Ian
dc.contributor.authorHarris, Matthew
dc.contributor.authorHodgson, Rose
dc.contributor.authorShah, Archna
dc.contributor.authorDowell, Simon J
dc.contributor.authorMobarec, Juan Carlos
dc.contributor.authorWoodlock, David A
dc.contributor.authorReynolds, Christopher A
dc.contributor.authorPoyner, David R
dc.contributor.authorWatkins, Harriet A
dc.contributor.authorLadds, Graham
dc.date.accessioned2016-10-21T09:11:11Z
dc.date.available2016-10-21T09:11:11Z
dc.date.issued2016-10-14
dc.identifier.issn0021-9258
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/260854
dc.description.abstractThe 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.
dc.description.sponsorshipThis work was supported by the National Heart Foundation of New Zealand (H.W.), the School of Biological Sciences, University of Auckland seed fund (H.W.), the BBSRC (G.L. - BB/M00015X/1), (D.P. - BB/M000176/1), (C.A.R. - BB/M006883/1), a BBSRC Doctoral Training Partnership (M.H. – BB/JO14540/1), an MRC Doctoral Training Partnership (I.W. - MR/J003964/1), a Warwick Impact Fund (C.W., G.L.), a Warwick Research Development Fund (C.W., G.L.) grant number (RD13301) and the Warwick Undergraduate Research Scholarship Scheme (A.S and R.H).
dc.languageEnglish
dc.language.isoen
dc.publisherElsevier BV
dc.subjectCGRP
dc.subjectadrenomedullin
dc.subjectadrenomedullin 2
dc.subjectG protein-coupled receptors (GPCRs)
dc.subjectreceptor activity modifying proteins (RAMPs)
dc.subjectsignal bias
dc.subjectsignal transduction
dc.subjectyeast
dc.subjectmolecular modelling
dc.subjectmolecular dynamics
dc.titleReceptor Activity-modifying Protein-directed G Protein Signaling Specificity for the Calcitonin Gene-related Peptide Family of Receptors.
dc.typeArticle
dc.description.versionThis is the author accepted manuscript. It is currently under an indefinite embargo pending publication by the American Society for Biochemistry and Molecular Biology.
prism.publicationDate2016
prism.publicationNameJ Biol Chem
dc.identifier.doi10.17863/CAM.6014
dcterms.dateAccepted2016-08-22
rioxxterms.versionofrecord10.1074/jbc.M116.751362
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2016
dc.contributor.orcidHarris, Matthew [0000-0002-7918-5735]
dc.contributor.orcidLadds, Graham [0000-0001-7320-9612]
dc.identifier.eissn1083-351X
rioxxterms.typeJournal Article/Review
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/M00015X/2)
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (1643678)
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/M00015X/1)
cam.issuedOnline2016-08-26
cam.orpheus.successThu Jan 30 12:57:32 GMT 2020 - The item has an open VoR version.
rioxxterms.freetoread.startdate2100-01-01


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