Repository logo
 
Publication

Live-cell FRET imaging reveals clustering of the prion protein at the cell surface induced by infectious prions

2014-07Journal article Open access
Simple item page
dc.contributor.authorTavares, Evandro
dc.contributor.authorMacedo, J.A.
dc.contributor.authorPaulo, Pedro M. R.
dc.contributor.authorSousa, Catarina
dc.contributor.authorLopes, Carlos
dc.contributor.authorMelo, Eduardo
dc.date.accessioned2018-12-07T14:53:20Z
dc.date.available2018-12-07T14:53:20Z
dc.date.issued2014-07
dc.description.abstractPrion diseases are associated to the conversion of the prion protein into a misfolded pathological isoform. The mechanism of propagation of protein misfolding by protein templating remains largely unknown. Neuroblastoma cells were transfected with constructs of the prion protein fused to both CFP-GPI-anchored and to YFP-GPI-anchored and directed to its cell membrane location. Live-cell FRET imaging between the prion protein fused to CFP or YFP was measured giving consistent values of 10 +/- 2%. This result was confirmed by fluorescence lifetime imaging microscopy and indicates intermolecular interactions between neighbor prion proteins. In particular, considering that a maximum FRET efficiency of 17 +/- 2% was determined from a positive control consisting of a fusion CFP-YFP-GPI-anchored. A stable cell clone expressing the two fusions containing the prion protein was also selected to minimize cell-to-cell variability. In both, stable and transiently transfected cells, the FRET efficiency consistently increased in the presence of infectious prions - from 4 +/- 1% to 7 +/- 1% in the stable clone and from 10 +/- 2% to 16 +/- 1% in transiently transfected cells. These results clearly reflect an increased clustering of the prion protein on the membrane in the presence of infectious prions, which was not observed in negative control using constructs without the prion protein and upon addition of non-infected brain. Our data corroborates the recent view that the primary site for prion conversion is the cell membrane. Since our fluorescent cell clone is not susceptible to propagate infectivity, we hypothesize that the initial event of prion infectivity might be the clustering of the GPI-anchored prion protein. (C) 2014 Elsevier B.V. All rights reserved.
dc.description.sponsorshipFundacao para a Ciencia e Tecnologia (FCT), Portugal [PTDC/QUI-BIQ/119677/2010, PTDC/CTM-NAN/2700/2012]; FCT [PEst-OE/EQB/LA0023/2011, SFRH/BD/48664/2008, SFRH/BPD/64932/2009]
dc.description.versioninfo:eu-repo/semantics/publishedVersion
dc.identifier.doi10.1016/j.bbadis.2014.02.002
dc.identifier.issn0925-4439
dc.identifier.issn0006-3002
dc.identifier.urihttp://hdl.handle.net/10400.1/11458
dc.language.isoeng
dc.peerreviewedyes
dc.publisherElsevier Science Bv
dc.relationFunctionalized Metal Nano-Gaps for Plasmon-Enhanced Single Protein Biosensing
dc.relationAN ANIMAL CELL-BASED APPROACH TO STUDY PRION DISEASES: CELLULAR TRAFFICKING, CONFORMATIONAL CHANGES AND PRION INFECTIVITY
dc.relationFUNCTION AND INFECTIVITY OF PRIONS DURING NEURAL DIFFERENTIATION OF EMBRYONIC STEM CELLS
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectResonance energy-transfer
dc.subjectGpi-Anchored proteins
dc.subjectNeurodegenerative diseases
dc.subjectFluorescent proteins
dc.subjectSignal-transduction
dc.subjectScrapie isoform
dc.subjectLiving cells
dc.subjectLipid rafts
dc.subjectMicroscopy
dc.subjectMembrane
dc.titleLive-cell FRET imaging reveals clustering of the prion protein at the cell surface induced by infectious prions
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleFunctionalized Metal Nano-Gaps for Plasmon-Enhanced Single Protein Biosensing
oaire.awardTitleAN ANIMAL CELL-BASED APPROACH TO STUDY PRION DISEASES: CELLULAR TRAFFICKING, CONFORMATIONAL CHANGES AND PRION INFECTIVITY
oaire.awardTitleFUNCTION AND INFECTIVITY OF PRIONS DURING NEURAL DIFFERENTIATION OF EMBRYONIC STEM CELLS
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FQUI-BIQ%2F119677%2F2010/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FCTM-NAN%2F2700%2F2012/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBD%2F48664%2F2008/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBPD%2F64932%2F2009/PT
oaire.citation.endPage991
oaire.citation.issue7
oaire.citation.startPage981
oaire.citation.titleBiochimica et Biophysica Acta-Molecular Basis of Disease
oaire.citation.volume1842
oaire.fundingStream3599-PPCDT
oaire.fundingStream3599-PPCDT
person.familyNameMacedo
person.familyNameSousa
person.familyNameLopes
person.familyNamePinho Melo
person.givenNameJoana
person.givenNameCatarina
person.givenNameCarlos
person.givenNameEduardo
person.identifier1443188
person.identifier.ciencia-idE812-421F-FF29
person.identifier.ciencia-id7914-48A2-AA34
person.identifier.ciencia-id5B12-908A-21A2
person.identifier.ciencia-id3C1C-C10C-1510
person.identifier.orcid0000-0001-9564-4770
person.identifier.orcid0000-0003-2608-6423
person.identifier.orcid0000-0002-6136-0478
person.identifier.orcid0000-0002-0974-8977
person.identifier.ridK-4979-2015
person.identifier.scopus-author-id56094485100
person.identifier.scopus-author-id35566177900
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccess
rcaap.typearticle
relation.isAuthorOfPublication285d1374-c1db-4862-b8d6-57bb2d0fb815
relation.isAuthorOfPublication64b13cc7-5212-4611-8af8-bfd5b8bfee8d
relation.isAuthorOfPublication21cb74bb-fd65-4da8-a95c-73d711c9c9d3
relation.isAuthorOfPublication5fa1895f-5577-4652-961a-886ec9bf41b1
relation.isAuthorOfPublication.latestForDiscovery64b13cc7-5212-4611-8af8-bfd5b8bfee8d
relation.isProjectOfPublication355a8acc-162a-4ab9-8628-cb6abfec6e02
relation.isProjectOfPublication87fb20a8-59b7-4fa1-8bdb-ef740af19bf6
relation.isProjectOfPublication9596af8b-f8af-4f6c-90a1-eb9bc1a053fe
relation.isProjectOfPublication9d202946-c800-467e-943c-96becd56869d
relation.isProjectOfPublication.latestForDiscovery87fb20a8-59b7-4fa1-8bdb-ef740af19bf6

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
H11458.pdf
Size:
1.06 MB
Format:
Adobe Portable Document Format
Download

Collections

FCB2-Artigos (em revistas ou actas indexadas)