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dc.contributor.authorWeston, Cathrynen
dc.contributor.authorWinfield, Ianen
dc.contributor.authorHarris, Matthewen
dc.contributor.authorHodgson, Roseen
dc.contributor.authorShah, Archnaen
dc.contributor.authorDowell, Simon Jen
dc.contributor.authorMobarec, Juan Carlosen
dc.contributor.authorWoodlock, David Aen
dc.contributor.authorReynolds, Christopher Aen
dc.contributor.authorPoyner, David Ren
dc.contributor.authorWatkins, Harriet Aen
dc.contributor.authorLadds, Grahamen
dc.date.accessioned2016-10-21T09:11:11Z
dc.date.available2016-10-21T09:11:11Z
dc.date.issued2016en
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 association of the calcitonin receptor-like receptor (CLR) and one of three receptor activitymodifying proteins (RAMPs). Binding of one of the three peptide ligands, CGRP, adrenomedullin (AM) or intermedin/adrenomedullin2 (AM2) is well known to result in a Gαs-mediated increase in cAMP. Here we use 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 Gα$_{s}$ and Gα$_{q}$ but also identify a Gα$_{i}$ 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 between Gα$_{s}$, Gα$_{i}$ and Gα$_{q/11}$ pathways. The results are discussed in the context of RAMP interactions probed through molecular modelling 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.languageEnglishen
dc.language.isoenen
dc.publisherAmerican Society for Biochemistry and Molecular Biology
dc.subjectCGRPen
dc.subjectadrenomedullinen
dc.subjectadrenomedullin 2en
dc.subjectG protein-coupled receptors (GPCRs)en
dc.subjectreceptor activity modifying proteins (RAMPs)en
dc.subjectsignal biasen
dc.subjectsignal transductionen
dc.subjectyeasten
dc.subjectmolecular modellingen
dc.subjectmolecular dynamicsen
dc.titleReceptor activity modifying protein-directed G protein signaling specificity for the calcitonin gene-related peptide family of receptorsen
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.en
prism.publicationDate2016en
prism.publicationNameThe Journal of Biological Chemistryen
dc.identifier.doi10.17863/CAM.6014
dcterms.dateAccepted2016-08-22en
rioxxterms.versionAMen
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2016en
dc.contributor.orcidHarris, Matthew [0000-0002-7918-5735]
dc.contributor.orcidLadds, Graham [0000-0001-7320-9612]
dc.identifier.eissn1083-351X
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idBBSRC (BB/M00015X/2)
pubs.funder-project-idBBSRC (1643678)
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/M00015X/1)
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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