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A fluorogenic probe for cell surface phosphatidylserine using an intramolecular indicator displacement sensing mechanism

2018Journal article Restricted access
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dc.contributor.authorZwicker, Vincent E.
dc.contributor.authorOliveira, Bruno
dc.contributor.authorYeo, Jia Hao
dc.contributor.authorFraser, Stuart T.
dc.contributor.authorBernardes, Gonçalo J. L.
dc.contributor.authorNew, Elizabeth J.
dc.contributor.authorJolliffe, Katrina A.
dc.date.accessioned2021-12-10T16:21:38Z
dc.date.available2021-12-10T16:21:38Z
dc.date.issued2018
dc.description© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheimpt_PT
dc.description.abstractThe detection of externalized phosphatidylserine (PS) on the cell surface is commonly used to distinguish between living, apoptotic, and necrotic cells. The tools of choice for many researchers to study apoptosis are annexin V-fluorophore conjugates. However, the use of this 35 kDa protein is associated with several drawbacks, including temperature sensitivity, Ca2+ dependence, and slow binding kinetics. Herein, a fluorogenic probe for cell surface PS, P-IID, is described, which operates by an intramolecular indicator displacement (IID) mechanism. An intramolecularly bound coumarin indicator is released in the presence of cell surface PS, leading to a fluorescence "turn-on" response. P-IID demonstrates superior performance when compared to annexin V, for both fluorescence imaging and flow cytometry. This allows P-IID to be used in time-lapse imaging of apoptosis using confocal laser scanning microscopy and demonstrates the utility of the IID mechanism in live cells.pt_PT
dc.description.sponsorshipWe thank the Australian Research Council (DP140100227 to K.A.J., and DP180101353 to K.A.J. and E.J.N.); the University of Sydney (IPRS and John A. Lamberton scholarships to V.E.Z.; P rof. N .G.W. and Mrs. Ann Macintosh Memorial scholarship to J.H.Y.; NWG Macintosh Grant 2018 to S.T.F.); the Royal Society (URF to G.J.L.B., UF110046 and URF\R\180019), FCT Portugal (iFCT to G.J.L.B., IF/00624/2015), ERC StG (GA No. 676832), the European Commission (Marie Sklodowska-Curie ITN ProteinConjugates GA No. 675007 to G.J.L.B. and Marie Sklodowska-Curie IEF GA No. 702574 to B.L.O.) and the EPSRC (EP/M003647/1 to G.J.L.B.) for funding. We also acknowledge the Australian Centre for Microscopy & Microanalysis (ACMM) and the Bosch Live Cell Analysis Facility (LCAF) for access to facilitiespt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationAngew Chem Int Ed Engl. 2019 Mar 4;58(10):3087-3091pt_PT
dc.identifier.doi10.1002/anie.201812489pt_PT
dc.identifier.eissn1521-3773
dc.identifier.issn1433-7851
dc.identifier.urihttp://hdl.handle.net/10451/50347
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherWileypt_PT
dc.relationIF/00624/2015pt_PT
dc.relationA Minimal-Tag Bioorthogonal Labelling Approach to Protein Uptake, Traffic and Delivery
dc.relationDevelopment of a cell-based system for high-throughput screening of antifibrotics
dc.relation.publisherversionhttps://onlinelibrary.wiley.com/journal/15213773pt_PT
dc.subjectAnnexin Vpt_PT
dc.subjectApoptosispt_PT
dc.subjectFluorescent probespt_PT
dc.subjectImaging agentspt_PT
dc.subjectPhosphatidylserinept_PT
dc.titleA fluorogenic probe for cell surface phosphatidylserine using an intramolecular indicator displacement sensing mechanismpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleA Minimal-Tag Bioorthogonal Labelling Approach to Protein Uptake, Traffic and Delivery
oaire.awardTitleDevelopment of a cell-based system for high-throughput screening of antifibrotics
oaire.awardURIinfo:eu-repo/grantAgreement/EC/H2020/676832/EU
oaire.awardURIinfo:eu-repo/grantAgreement/EC/H2020/702574/EU
oaire.citation.endPage3091pt_PT
oaire.citation.issue10pt_PT
oaire.citation.startPage3087pt_PT
oaire.citation.titleAngewandte Chemie International Editionpt_PT
oaire.citation.volume58pt_PT
oaire.fundingStreamH2020
oaire.fundingStreamH2020
person.familyNameOliveira
person.familyNameBernardes
person.givenNameBruno
person.givenNameGonçalo
person.identifier.ciencia-idA016-1DDF-E7CB
person.identifier.orcid0000-0002-7687-4746
person.identifier.orcid0000-0001-6594-8917
person.identifier.ridT-5374-2018
person.identifier.scopus-author-id24577505300
person.identifier.scopus-author-id14046757500
project.funder.identifierhttp://doi.org/10.13039/501100008530
project.funder.identifierhttp://doi.org/10.13039/501100008530
project.funder.nameEuropean Commission
project.funder.nameEuropean Commission
rcaap.rightsrestrictedAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication18a3cb41-d651-4f5e-acb1-1ffd4dfeffb1
relation.isAuthorOfPublicationd1a48067-77b1-4413-b1c7-602fb18c62c0
relation.isAuthorOfPublication.latestForDiscovery18a3cb41-d651-4f5e-acb1-1ffd4dfeffb1
relation.isProjectOfPublication2f566f67-6fd2-4a9a-a4f4-b2864d98ef5f
relation.isProjectOfPublication9501df5c-799b-4d9f-8d11-bbbf57ca2749
relation.isProjectOfPublication.latestForDiscovery2f566f67-6fd2-4a9a-a4f4-b2864d98ef5f

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