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To what extent do fluorophores bias the biological activity of peptides? A practical approach using membrane-active peptides as models

2020Journal article Open access
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dc.contributor.authorCavaco, Marco
dc.contributor.authorPérez-Peinado, Clara
dc.contributor.authorValle, Javier
dc.contributor.authorSilva, Rúben
dc.contributor.authorCorreia, João D. G.
dc.contributor.authorCastanho, Miguel A. R. B.
dc.contributor.authorNeves, Vera
dc.contributor.authorAndreu, David
dc.date.accessioned2023-05-17T14:31:19Z
dc.date.available2023-05-17T14:31:19Z
dc.date.issued2020
dc.descriptionCopyright © 2020 Cavaco, Pérez-Peinado, Valle, Silva, Correia, Andreu, Castanho and Neves. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these termspt_PT
dc.description.abstractThe characterization of biologically active peptides relies heavily on the study of their efficacy, toxicity, mechanism of action, cellular uptake, or intracellular location, using both in vitro and in vivo studies. These studies frequently depend on the use of fluorescence-based techniques. Since most peptides are not intrinsically fluorescent, they are conjugated to a fluorophore. The conjugation may interfere with peptide properties, thus biasing the results. The selection of the most suitable fluorophore is highly relevant. Here, a comprehensive study with blood–brain barrier (BBB) peptide shuttles (PepH3 and PepNeg) and antimicrobial peptides (AMPs) (vCPP2319 and Ctn[15-34]), tested as anticancer peptides (ACPs), having different fluorophores, namely 5(6)-carboxyfluorescein (CF), rhodamine B (RhB), quasar 570 (Q570), or tide fluor 3 (TF3) attached is presented. The goal is the evaluation of the impact of the selected fluorophores on peptide performance, applying routinely used techniques to assess cytotoxicity/toxicity, secondary structure, BBB translocation, and cellular internalization. Our results show that some fluorophores significantly modulate peptide activity when compared with unlabeled peptides, being more noticeable in hydrophobic and charged fluorophores. This study highlights the need for a careful experimental design for fluorescently labeled molecules, such as peptides.pt_PT
dc.description.sponsorshipThe authors thank the Portuguese Funding Agency, Fundação para a Ciência e a Tecnologia, FCT IP, for financial support (grants: PD/BD/128281/2017, PTDC/BBB-NAN/1578/2014); European Union’s Horizon 2020 Research and Innovation Programme under grant agreement No 828774; and “la Caixa” Banking Foundation under the project code HR17-00409.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationFrontiers in Bioengineering and Biotechnology, September 2020 | Volume 8 | Article 552035pt_PT
dc.identifier.doi10.3389/fbioe.2020.552035pt_PT
dc.identifier.eissn2296-4185
dc.identifier.urihttp://hdl.handle.net/10451/57440
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherFrontiers Mediapt_PT
dc.relationTrans-BBB peptides for targeting brain metastasis
dc.relation''One size fits all'' unique drug to eradicate multiple viral species simultaneously from the central nervous system of co-infected individuals
dc.relation.publisherversionhttps://www.frontiersin.org/journals/bioengineering-and-biotechnologypt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectAnticancer peptidespt_PT
dc.subjectBBB peptide shuttlespt_PT
dc.subjectFluorescencept_PT
dc.subjectFluorophorept_PT
dc.subjectLabelingpt_PT
dc.titleTo what extent do fluorophores bias the biological activity of peptides? A practical approach using membrane-active peptides as modelspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleTrans-BBB peptides for targeting brain metastasis
oaire.awardTitle''One size fits all'' unique drug to eradicate multiple viral species simultaneously from the central nervous system of co-infected individuals
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//PD%2FBD%2F128281%2F2017/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FBBB-NAN%2F1578%2F2014/PT
oaire.awardURIinfo:eu-repo/grantAgreement/EC/H2020/828774/EU
oaire.citation.startPage552035pt_PT
oaire.citation.titleFrontiers in Bioengineering and Biotechnologypt_PT
oaire.citation.volume8pt_PT
oaire.fundingStream3599-PPCDT
oaire.fundingStreamH2020
person.familyNameCavaco
person.familyNameSilva
person.familyNameCorreia
person.familyNameCastanho
person.familyNameNeves
person.givenNameMarco
person.givenNameRúben
person.givenNameJoão
person.givenNameMiguel
person.givenNameVera
person.identifier1069324
person.identifier1064683
person.identifier953259
person.identifier.ciencia-id1412-63B8-7494
person.identifier.ciencia-id561A-878D-536F
person.identifier.ciencia-idEC11-F3E9-78A2
person.identifier.ciencia-id671C-1860-A160
person.identifier.orcid0000-0002-0938-9038
person.identifier.orcid0000-0003-3665-9571
person.identifier.orcid0000-0002-7847-4906
person.identifier.orcid0000-0001-7891-7562
person.identifier.orcid0000-0002-2989-7208
person.identifier.ridJ-7036-2013
person.identifier.ridO-2176-2018
person.identifier.scopus-author-id7202364104
person.identifier.scopus-author-id56605575600
person.identifier.scopus-author-id26537945300
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100008530
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameEuropean Commission
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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