Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry

Detalhes bibliográficos
Autor(a) principal: Freire, João Miguel
Data de Publicação: 2015
Outros Autores: Gaspar, Diana, Torre, Beatriz Garcia de la, Veiga, Ana Salomé, Andreu, David, Castanho, Miguel A. R. B.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10451/23430
Resumo: © 2014 Elsevier B.V. All rights reserved
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spelling Monitoring antibacterial permeabilization in real time using time-resolved flow cytometryTime-resolved flow cytometryAntimicrobial peptideLive/dead assayPeptide–lipid interactionAntimicrobial actionLipopolysaccharide© 2014 Elsevier B.V. All rights reservedDespite the intensive study of antibiotic-induced bacterial permeabilization, its kinetics and molecular mechanism remain largely elusive. A new methodology that extends the concept of the live–dead assay in flow cytometry to real time-resolved detection was used to overcome these limitations. The antimicrobial activity of pepR was monitored in time-resolved flow cytometry for three bacterial strains: Escherichia coli (ATCC 25922), E. coli K-12 (CGSC Strain 4401) and E. coli JW3596-1 (CGSC Strain 11805). The latter strain has truncated lipopolysaccharides (LPS) in the outer membrane. This new methodology provided information on the efficacy of the antibiotics and sheds light on their mode of action atmembrane-level. Kinetic data regarding antibiotic binding and lytic actionwere retrieved.Membrane interaction and permeabilization events differ significantly among strains. The truncation of LPS moieties does not hamper AMP binding but compromises membrane disruption and bacterial killing. We demonstrated the usefulness of time-resolved flow cytometry to study antimicrobial-induced permeabilization by collecting kinetic data that contribute to characterize the action of antibiotics directly on bacteria.We thank CGSC, Coli Genetic Stock Center, for the K-12 and JW3596-1 Escherichia coli strains. This work was supported by project PTDC/QUIBIQ/112929/2009 from Fundação para a Ciência e a Tecnologia–Ministério da Educação e Ciência (FCT-MEC, Portugal). JMF and DG also acknowledge FCT-MEC for fellowships SFRH/BD/70423/2010 and SFRH/BPD/73500/2010, respectively, and ASV for funding within the FCT Investigator Programme (IF/00803/2012). Work at Pompeu Fabra University was supported by grants from the Spanish Ministry of Economy and Competitiveness (SAF2011-24899) and from Generalitat de Catalunya (SGR2009-0492).ElsevierRepositório da Universidade de LisboaFreire, João MiguelGaspar, DianaTorre, Beatriz Garcia de laVeiga, Ana SaloméAndreu, DavidCastanho, Miguel A. R. B.2016-04-18T16:01:06Z20152015-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10451/23430engBiochimica et Biophysica Acta 1848 (2015) 554–5600005-273610.1016/j.bbamem.2014.11.001metadata only accessinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-08T16:10:48Zoai:repositorio.ul.pt:10451/23430Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T21:40:31.369258Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry
title Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry
spellingShingle Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry
Freire, João Miguel
Time-resolved flow cytometry
Antimicrobial peptide
Live/dead assay
Peptide–lipid interaction
Antimicrobial action
Lipopolysaccharide
title_short Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry
title_full Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry
title_fullStr Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry
title_full_unstemmed Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry
title_sort Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry
author Freire, João Miguel
author_facet Freire, João Miguel
Gaspar, Diana
Torre, Beatriz Garcia de la
Veiga, Ana Salomé
Andreu, David
Castanho, Miguel A. R. B.
author_role author
author2 Gaspar, Diana
Torre, Beatriz Garcia de la
Veiga, Ana Salomé
Andreu, David
Castanho, Miguel A. R. B.
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Repositório da Universidade de Lisboa
dc.contributor.author.fl_str_mv Freire, João Miguel
Gaspar, Diana
Torre, Beatriz Garcia de la
Veiga, Ana Salomé
Andreu, David
Castanho, Miguel A. R. B.
dc.subject.por.fl_str_mv Time-resolved flow cytometry
Antimicrobial peptide
Live/dead assay
Peptide–lipid interaction
Antimicrobial action
Lipopolysaccharide
topic Time-resolved flow cytometry
Antimicrobial peptide
Live/dead assay
Peptide–lipid interaction
Antimicrobial action
Lipopolysaccharide
description © 2014 Elsevier B.V. All rights reserved
publishDate 2015
dc.date.none.fl_str_mv 2015
2015-01-01T00:00:00Z
2016-04-18T16:01:06Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10451/23430
url http://hdl.handle.net/10451/23430
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Biochimica et Biophysica Acta 1848 (2015) 554–560
0005-2736
10.1016/j.bbamem.2014.11.001
dc.rights.driver.fl_str_mv metadata only access
info:eu-repo/semantics/openAccess
rights_invalid_str_mv metadata only access
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
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reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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