Show simple item record

dc.contributor.authorBailey, Sian
dc.contributor.authorHarris, Matthew
dc.contributor.authorBarkan, Kerry
dc.contributor.authorWinfield, Ian
dc.contributor.authorHarper, Matthew Thomas
dc.contributor.authorSimms, John
dc.contributor.authorLadds, Graham
dc.contributor.authorWheatley, Mark
dc.contributor.authorPoyner, David
dc.date.accessioned2019-03-05T00:31:26Z
dc.date.available2019-03-05T00:31:26Z
dc.date.issued2019-05-01
dc.identifier.issn0005-2736
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/290194
dc.description.abstractCorticotrophin releasing factor (CRF) acts via two family B G-protein-coupled receptors, CRFR1 and CRFR2. Additional subtypes exist due to alternative splicing. CRFR1α is the most widely expressed subtype and lacks a 29-residue insert in the first intracellular loop that is present in CRFR1β. It has been shown previously that co-expression of CRFR1β with receptor activity modifying protein 2 (RAMP2) in HEK 293S cells increased the cell-surface expression of both proteins suggesting a physical interaction as seen with RAMPs and calcitonin receptor-like receptor (CLR). This study investigated the ability of CRFR1α, CRFR1β and CRFR2β to promote cell-surface expression of FLAG-tagged RAMP2. Four different cell-lines were utilised to investigate the effect of varying cellular context; COS-7, HEK 293T, HEK 293S and [ΔCTR]HEK 293 (which lacks endogenous calcitonin receptor). In all cell-lines, CRFR1α and CRFR1β enhanced RAMP2 cell-surface expression. The magnitude of the effect on RAMP2 was dependent on the cell-line ([ΔCTR]HEK 293 > COS-7 > HEK 293T > HEK 293S). RT-PCR indicated this variation may relate to differences in endogenous RAMP expression between cell types. Furthermore, pre-treatment with CRF resulted in a loss of cell-surface FLAG-RAMP2 when it was co-expressed with CRFR1 subtypes. CRFR2β co-expression had no effect on RAMP2 in any cell-line. Molecular modelling suggests that the potential contact interface between the extracellular domains of RAMP2 and CRF receptor subtypes is smaller than that of RAMP2 and CRL, the canonical receptor:RAMP pairing, assuming a physical interaction. Furthermore, a specific residue difference between CRFR1 subtypes (glutamate) and CRFR2β (histidine) in this interface region may impair CRFR2β:RAMP2 interaction by electrostatic repulsion.
dc.format.mediumPrint-Electronic
dc.languageeng
dc.publisherElsevier BV
dc.subjectCOS Cells
dc.subjectAnimals
dc.subjectHumans
dc.subjectReceptors, Corticotropin-Releasing Hormone
dc.subjectAlternative Splicing
dc.subjectModels, Molecular
dc.subjectHEK293 Cells
dc.subjectReceptor Activity-Modifying Protein 2
dc.subjectChlorocebus aethiops
dc.titleInteractions between RAMP2 and CRF receptors: The effect of receptor subtypes, splice variants and cell context.
dc.typeArticle
prism.endingPage1003
prism.issueIdentifier5
prism.publicationDate2019
prism.publicationNameBiochim Biophys Acta Biomembr
prism.startingPage997
prism.volume1861
dc.identifier.doi10.17863/CAM.37421
dcterms.dateAccepted2019-02-26
rioxxterms.versionofrecord10.1016/j.bbamem.2019.02.008
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2019-05
dc.contributor.orcidHarris, Matthew [0000-0002-7918-5735]
dc.contributor.orcidHarper, Matthew [0000-0002-4740-637X]
dc.contributor.orcidLadds, Graham [0000-0001-7320-9612]
dc.identifier.eissn1879-2642
rioxxterms.typeJournal Article/Review
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/M00015X/2)
pubs.funder-project-idLeverhulme Trust (via University of Essex) (DBG3000)
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (1643678)
cam.issuedOnline2019-02-28
cam.orpheus.successThu Jan 30 10:49:24 GMT 2020 - Embargo updated
rioxxterms.freetoread.startdate2020-05-31


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record