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Fluorescent Leakage Assay to Investigate Membrane Destabilization by Cell-Penetrating Peptide
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Daptomycin Leakage Is Selective.

Jin Zhang1, Kyle Scoten1, Suzana K Straus1

  • 1Department of Chemistry, University of British Columbia , 2036 Main Mall, Vancouver, British Columbia V6T 1Z1, Canada.

ACS Infectious Diseases
|September 28, 2016
PubMed
Summary
This summary is machine-generated.

Daptomycin antibiotic action was investigated using model membranes. A new robust system suggests daptomycin selectively targets potassium ions, advancing understanding of its antibacterial mechanism.

Keywords:
1,3-benzenedicarboxylic acid4,4′-[1,4,10,13-tetraoxa-7,16-diazacyclooctadecane-7,16-diylbis(5-methoxy-6,2-benzofurandiyl)]bis-, tetrakis[(acetyloxy)methyl] ester (PBFI)4,4′-[1,4,10-trioxa-7,13-diazacyclopentadecane-7,13-diylbis(5-methoxy-6,2-benzofurandiyl)]bis-, tetraammonium salt (SBFI)carbonyl cyanide m-chlorophenyl hydrazine (CCCP)daptomycinfluorescenceleakagelipopeptidemembrane depolarizationpyranine

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Area of Science:

  • Biochemistry
  • Microbiology
  • Membrane Biophysics

Background:

  • Daptomycin is a lipopeptide antibiotic effective against Gram-positive bacteria.
  • Its mechanism of action is proposed to involve membrane permeabilization and depolarization.
  • Previous models used specific lipid compositions and ionophores to study daptomycin's effects.

Purpose of the Study:

  • To investigate the dependence of a model membrane system on composition and the role of CCCP in studying daptomycin's mode of action.
  • To evaluate a more robust model membrane system for investigating daptomycin's interaction with bacterial membranes.
  • To determine the ion selectivity of daptomycin's membrane interaction.

Main Methods:

  • Utilized a fluorescence assay with pyranine to assess liposome leakage.
  • Employed a novel model membrane system incorporating ion-selective dyes PBFI and SBFI.
  • Analyzed daptomycin's effect on membrane stability and ion permeability.

Main Results:

  • Daptomycin induced leakage in liposomes of limited stability.
  • The ionophore CCCP was found to enhance daptomycin-induced leakage.
  • The PBFI/SBFI model demonstrated that daptomycin exhibits selectivity for potassium ions.

Conclusions:

  • The previously proposed model membrane system has limitations regarding stability and CCCP dependence.
  • A new model membrane system using PBFI and SBFI offers a more robust approach.
  • Daptomycin's antibacterial activity is likely mediated by selective disruption of potassium ion transport across the bacterial membrane.