Utilize este identificador para referenciar este registo: http://hdl.handle.net/10451/23430
Título: Monitoring antibacterial permeabilization in real time using time-resolved flow cytometry
Autor: Freire, João Miguel
Gaspar, Diana
Torre, Beatriz Garcia de la
Veiga, Ana Salomé
Andreu, David
Castanho, Miguel A. R. B.
Palavras-chave: Time-resolved flow cytometry
Antimicrobial peptide
Live/dead assay
Peptide–lipid interaction
Antimicrobial action
Data: 2015
Editora: Elsevier
Citação: Biochimica et Biophysica Acta 1848 (2015) 554–560
Resumo: 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 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.
Descrição: © 2014 Elsevier B.V. All rights reserved
Peer review: yes
URI: http://hdl.handle.net/10451/23430
DOI: 10.1016/j.bbamem.2014.11.001
ISSN: 0005-2736
Versão do Editor: http://www.sciencedirect.com/science/article/pii/S0005273614003770
Aparece nas colecções:IMM - Artigos em Revistas Internacionais

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