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dc.contributor.authorInduruwa, Isuruen
dc.contributor.authorMoroi, Men
dc.contributor.authorBonna, Arkadiuszen
dc.contributor.authorMalcor, J-Den
dc.contributor.authorDear, Joanna-Marieen
dc.contributor.authorWarburton, Lizen
dc.contributor.authorFarndale, Richarden
dc.contributor.authorJung, Stephanieen
dc.date.accessioned2018-02-09T09:52:47Z
dc.date.available2018-02-09T09:52:47Z
dc.date.issued2018-02en
dc.identifier.issn1538-7933
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/271845
dc.description.abstractSummary. Background: Platelet collagen receptor GPVI binds collagen, initiating thrombogenesis and stabilizes thrombi by binding fibrin. Objectives: To determine if GPVI-dimer, monomer, or both bind to fibrinogen substrates, and which region common to these substrates contains the interaction site. Methods: Recombinant GPVI monomeric extracellular domain (GPVIex) or dimeric Fc-fusion protein (GPVI-Fc2) binding to immobilized fibrinogen derivatives was measured by ELISA, including competition assays involving collagenous substrates and fibrinogen derivatives. Flow adhesion was performed with normal or Glanzmann’s thrombasthenic (GT) platelets over immobilized fibrinogen, with or without anti-GPVI-dimer or anti-αIIbβ3. Results: Under static conditions, GPVIex did not bind to any fibrinogen substrate. GPVI-Fc2 exhibited specific, saturable binding to both D-fragment and D-dimer, which was inhibited by mFab-F (anti-GPVI-dimer), but showed low binding to fibrinogen and fibrin under our conditions. GPVI-Fc2 binding to D-fragment or D-dimer was abrogated by collagen type III, Horm collagen, or CRP-XL, suggesting proximity between the D-domain and collagen binding sites on GPVI-dimer. Under low shear, adhesion of normal platelets to D-fragment, D-dimer, fibrinogen and fibrin was inhibited by mFab-F and abolished by Eptifibatide (inhibitor of αIIbβ3), suggesting that both receptors contribute to thrombus formation on these substrates, but αIIbβ3 makes a greater contribution. Notably, thrombasthenic platelets showed limited adhesion to fibrinogen substrates under flow, which was further reduced by mFab-F, supporting some independent GPVI-dimer involvement in this interaction. Conclusion: Only dimeric GPVI interacts with fibrinogen D-domain, at a site proximate to its collagen binding site, to support platelet adhesion/activation/aggregate formation on immobilized fibrinogen and polymerized fibrin.
dc.format.mediumPrint-Electronicen
dc.languageengen
dc.publisherWiley-Blackwell
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjectBlood Plateletsen
dc.subjectHumansen
dc.subjectThrombosisen
dc.subjectThrombastheniaen
dc.subjectCollagenen
dc.subjectFibrinogenen
dc.subjectFibrinen
dc.subjectFibrin Fibrinogen Degradation Productsen
dc.subjectPlatelet Membrane Glycoproteinsen
dc.subjectPlatelet Glycoprotein GPIIb-IIIa Complexen
dc.subjectCase-Control Studiesen
dc.subjectSignal Transductionen
dc.subjectBinding Sitesen
dc.subjectProtein Bindingen
dc.subjectStructure-Activity Relationshipen
dc.subjectPlatelet Adhesivenessen
dc.subjectPlatelet Activationen
dc.subjectProtein Interaction Domains and Motifsen
dc.subjectProtein Multimerizationen
dc.titlePlatelet collagen receptor Glycoprotein VI-dimer recognizes fibrinogen and fibrin through their D-domains, contributing to platelet adhesion and activation during thrombus formation.en
dc.typeArticle
prism.endingPage404
prism.issueIdentifier2en
prism.publicationDate2018en
prism.publicationNameJournal of thrombosis and haemostasis : JTHen
prism.startingPage389
prism.volume16en
dc.identifier.doi10.17863/CAM.18852
dcterms.dateAccepted2017-11-26en
rioxxterms.versionofrecord10.1111/jth.13919en
rioxxterms.versionVoR*
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2018-02en
dc.contributor.orcidInduruwa, Isuru [0000-0002-7020-8179]
dc.contributor.orcidBonna, Arkadiusz [0000-0002-3957-7849]
dc.contributor.orcidJung, Stephanie [0000-0002-7409-9715]
dc.identifier.eissn1538-7836
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idBritish Heart Foundation (PG/10/011/28199)
pubs.funder-project-idBritish Heart Foundation (RG/15/4/31268)
pubs.funder-project-idBritish Heart Foundation (SP/15/7/31561)
pubs.funder-project-idBritish Heart Foundation (PG/10/011/28199)
cam.orpheus.successThu Jan 30 12:59:07 GMT 2020 - The item has an open VoR version.*
rioxxterms.freetoread.startdate2100-01-01


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International