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Title: The mechanism of action of pepR, a viral-derived peptide, against Staphylococcus aureus biofilms

Abstract

Abstract Objectives To investigate the mechanism of action at the molecular level of pepR, a multifunctional peptide derived from the Dengue virus capsid protein, against Staphylococcus aureus biofilms. Methods Biofilm mass, metabolic activity and viability were quantified using conventional microbiology techniques, while fluorescence imaging methods, including a real-time calcein release assay, were employed to investigate the kinetics of pepR activity at different biofilm depths. Results Using flow cytometry-based assays, we showed that pepR is able to prevent staphylococcal biofilm formation due to a fast killing of planktonic bacteria, which in turn resulted from a peptide-induced increase in the permeability of the bacterial membranes. The activity of pepR against pre-formed biofilms was evaluated through the application of a quantitative live/dead confocal laser scanning microscopy (CLSM) assay. The results show that the bactericidal activity of pepR on pre-formed biofilms is dose and depth dependent. A CLSM-based assay of calcein release from biofilm-embedded bacteria was further developed to indirectly assess the diffusion and membrane permeabilization properties of pepR throughout the biofilm. A slower diffusion and delayed activity of the peptide at deeper layers of the biofilm were quantified. Conclusions Overall, our results show that the activity of pepR on pre-formed biofilms is controlledmore » by its diffusion along the biofilm layers, an effect that can be counteracted by an additional administration of peptide. Our study sheds new light on the antibiofilm mechanism of action of antimicrobial peptides, particularly the importance of their diffusion properties through the biofilm matrix on their activity.« less

Authors:
 [1];  [2];  [2];  [3];  [1];  [4];  [4];  [1];  [2];  [5];  [2]
  1. Centro de Química-Física Molecular e IN, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais Lisboa, Portugal, iBB-Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais Lisboa, Portugal
  2. Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Av. Prof. Egas Moniz, Lisboa, Portugal
  3. iBB-Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais Lisboa, Portugal
  4. Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Barcelona Biomedical Research Park, Barcelona, Spain
  5. Centro de Química-Física Molecular e IN, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais Lisboa, Portugal, iBB-Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais Lisboa, Portugal, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande Lisboa, Portugal
Publication Date:
Sponsoring Org.:
USDOE Office of Nuclear Energy (NE), Fuel Cycle Technologies (NE-5)
OSTI Identifier:
1564567
Grant/Contract Number:  
SFRH/BPD/92409/2013; PD/BD/114425/2016; PD/BD/136866/2018; SFRH/BPD/91831/2012; IF/00803/2012; IF/00386/2015
Resource Type:
Published Article
Journal Name:
Journal of Antimicrobial Chemotherapy
Additional Journal Information:
Journal Name: Journal of Antimicrobial Chemotherapy Journal Volume: 74 Journal Issue: 9; Journal ID: ISSN 0305-7453
Publisher:
Oxford University Press
Country of Publication:
United Kingdom
Language:
English

Citation Formats

Pinto, Sandra N., Dias, Susana A., Cruz, Ana F., Mil-Homens, Dalila, Fernandes, Fabio, Valle, Javier, Andreu, David, Prieto, Manuel, Castanho, Miguel A. R. B., Coutinho, Ana, and Veiga, Ana Salomé. The mechanism of action of pepR, a viral-derived peptide, against Staphylococcus aureus biofilms. United Kingdom: N. p., 2019. Web. doi:10.1093/jac/dkz223.
Pinto, Sandra N., Dias, Susana A., Cruz, Ana F., Mil-Homens, Dalila, Fernandes, Fabio, Valle, Javier, Andreu, David, Prieto, Manuel, Castanho, Miguel A. R. B., Coutinho, Ana, & Veiga, Ana Salomé. The mechanism of action of pepR, a viral-derived peptide, against Staphylococcus aureus biofilms. United Kingdom. doi:10.1093/jac/dkz223.
Pinto, Sandra N., Dias, Susana A., Cruz, Ana F., Mil-Homens, Dalila, Fernandes, Fabio, Valle, Javier, Andreu, David, Prieto, Manuel, Castanho, Miguel A. R. B., Coutinho, Ana, and Veiga, Ana Salomé. Fri . "The mechanism of action of pepR, a viral-derived peptide, against Staphylococcus aureus biofilms". United Kingdom. doi:10.1093/jac/dkz223.
@article{osti_1564567,
title = {The mechanism of action of pepR, a viral-derived peptide, against Staphylococcus aureus biofilms},
author = {Pinto, Sandra N. and Dias, Susana A. and Cruz, Ana F. and Mil-Homens, Dalila and Fernandes, Fabio and Valle, Javier and Andreu, David and Prieto, Manuel and Castanho, Miguel A. R. B. and Coutinho, Ana and Veiga, Ana Salomé},
abstractNote = {Abstract Objectives To investigate the mechanism of action at the molecular level of pepR, a multifunctional peptide derived from the Dengue virus capsid protein, against Staphylococcus aureus biofilms. Methods Biofilm mass, metabolic activity and viability were quantified using conventional microbiology techniques, while fluorescence imaging methods, including a real-time calcein release assay, were employed to investigate the kinetics of pepR activity at different biofilm depths. Results Using flow cytometry-based assays, we showed that pepR is able to prevent staphylococcal biofilm formation due to a fast killing of planktonic bacteria, which in turn resulted from a peptide-induced increase in the permeability of the bacterial membranes. The activity of pepR against pre-formed biofilms was evaluated through the application of a quantitative live/dead confocal laser scanning microscopy (CLSM) assay. The results show that the bactericidal activity of pepR on pre-formed biofilms is dose and depth dependent. A CLSM-based assay of calcein release from biofilm-embedded bacteria was further developed to indirectly assess the diffusion and membrane permeabilization properties of pepR throughout the biofilm. A slower diffusion and delayed activity of the peptide at deeper layers of the biofilm were quantified. Conclusions Overall, our results show that the activity of pepR on pre-formed biofilms is controlled by its diffusion along the biofilm layers, an effect that can be counteracted by an additional administration of peptide. Our study sheds new light on the antibiofilm mechanism of action of antimicrobial peptides, particularly the importance of their diffusion properties through the biofilm matrix on their activity.},
doi = {10.1093/jac/dkz223},
journal = {Journal of Antimicrobial Chemotherapy},
number = 9,
volume = 74,
place = {United Kingdom},
year = {2019},
month = {5}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.1093/jac/dkz223

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Works referenced in this record:

Bacterial biofilms: from the Natural environment to infectious diseases
journal, February 2004

  • Hall-Stoodley, Luanne; Costerton, J. William; Stoodley, Paul
  • Nature Reviews Microbiology, Vol. 2, Issue 2, p. 95-108
  • DOI: 10.1038/nrmicro821

Bacterial Biofilms: A Common Cause of Persistent Infections
journal, May 1999