Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry
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- dc.contributor.author Miguel Freire, Joãoca
- dc.contributor.author Gaspar, Dianaca
- dc.contributor.author García de la Torre, Beatrizca
- dc.contributor.author Veiga, Ana Saloméca
- dc.contributor.author Andreu Martínez, Davidca
- dc.contributor.author Castanho, Miguel A.R.B.ca
- dc.date.accessioned 2016-03-18T17:25:58Z
- dc.date.available 2016-03-18T17:25:58Z
- dc.date.issued 2015
- dc.description.abstract Despite 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 at membrane-level. Kinetic data regarding antibiotic binding and lytic action were 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.ca
- dc.description.sponsorship 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 DGalso 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).
- dc.format.mimetype application/pdfca
- dc.identifier.citation Guixà-González R, Javanainen M, Gómez-Soler M, Cordobilla B, Domingo JC, Sanz F et al. Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry. Biochimica et biophysica acta. 2015; 148(2): 554-560. DOI 10.1038/srep19839ca
- dc.identifier.doi http://dx.doi.org/10.1038/srep19839
- dc.identifier.issn 0006-3002
- dc.identifier.uri http://hdl.handle.net/10230/26018
- dc.language.iso engca
- dc.publisher Elsevierca
- dc.relation.ispartof Biochimica et biophysica acta. 2015; 148(2): 554-560
- dc.relation.projectID info:eu-repo/grantAgreement/ES/3PN/SAF2011-24899
- dc.rights © Elsevier This is the published version of an article http://dx.doi.org/10.1038/srep19839 that appeared in the journal Biochimica et biophysica acta. It is published in an Open Archive under an Elsevier user license. Details of this licence are available here: http://www.elsevier.com/about/open-access/open-access-policies/oa-license-policy/elsevier-user-license.ca
- dc.rights.accessRights info:eu-repo/semantics/openAccessca
- dc.subject.keyword Time-resolved flow cytometry
- dc.subject.keyword Antimicrobial peptide
- dc.subject.keyword Live/dead assay
- dc.subject.keyword Peptide–lipid interaction
- dc.subject.keyword Antimicrobial action
- dc.subject.keyword Lipopolysaccharide
- dc.subject.other Pèptidsca
- dc.title Monitoring antibacterial permeabilization in real time using time-resolved flow cytometryca
- dc.type info:eu-repo/semantics/articleca
- dc.type.version info:eu-repo/semantics/publishedVersionca