In the present work, we investigated the antifungal activity of two de novo designed, antimicrobial peptides VS2 and VS3, incorporating unnatural amino acid α,β-dehydrophenylalanine (ΔPhe). We observed that the low-hemolytic peptides could irreversibly inhibit the growth of various Candida species and multidrug resistance strains at MIC(80) values ranging from 15.62μM to 250μM. Synergy experiments showed that MIC(80) of the peptides was drastically reduced in combination with an antifungal drug fluconazole. The dye PI uptake assay was used to demonstrate peptide induced cell membrane permeabilization. Intracellular localization of the FITC-labeled peptides in Candida albicans was studied by confocal microscopy and FACS. Killing kinetics, PI uptake assay, and the intracellular presence of FITC-peptides suggested that growth inhibition is not solely a consequence of increased membrane permeabilization. We showed that entry of the peptide in Candida cells resulted in accumulation of reactive oxygen species (ROS) leading to cell necrosis. Morphological alteration in Candida cells caused by the peptides was visualized by electron microscopy. We propose that de novo designed VS2 and VS3 peptides have multiple detrimental effects on target fungi, which ultimately result in cell wall disruption and killing. Therefore, these peptides represent a good template for further design and development as antifungal agents.
%0 Journal Article
%1 Maurya2011Antifungal
%A Maurya, Indresh Kumar K.
%A Pathak, Sarika
%A Sharma, Monika
%A Sanwal, Hina
%A Chaudhary, Preeti
%A Tupe, Santosh
%A Deshpande, Mukund
%A Chauhan, Virander Singh S.
%A Prasad, Rajendra
%D 2011
%J Peptides
%K ros
%R 10.1016/j.peptides.2011.06.003
%T Antifungal activity of novel synthetic peptides by accumulation of reactive oxygen species (ROS) and disruption of cell wall against Candida albicans.
%U http://dx.doi.org/10.1016/j.peptides.2011.06.003
%X In the present work, we investigated the antifungal activity of two de novo designed, antimicrobial peptides VS2 and VS3, incorporating unnatural amino acid α,β-dehydrophenylalanine (ΔPhe). We observed that the low-hemolytic peptides could irreversibly inhibit the growth of various Candida species and multidrug resistance strains at MIC(80) values ranging from 15.62μM to 250μM. Synergy experiments showed that MIC(80) of the peptides was drastically reduced in combination with an antifungal drug fluconazole. The dye PI uptake assay was used to demonstrate peptide induced cell membrane permeabilization. Intracellular localization of the FITC-labeled peptides in Candida albicans was studied by confocal microscopy and FACS. Killing kinetics, PI uptake assay, and the intracellular presence of FITC-peptides suggested that growth inhibition is not solely a consequence of increased membrane permeabilization. We showed that entry of the peptide in Candida cells resulted in accumulation of reactive oxygen species (ROS) leading to cell necrosis. Morphological alteration in Candida cells caused by the peptides was visualized by electron microscopy. We propose that de novo designed VS2 and VS3 peptides have multiple detrimental effects on target fungi, which ultimately result in cell wall disruption and killing. Therefore, these peptides represent a good template for further design and development as antifungal agents.
@article{Maurya2011Antifungal,
abstract = {In the present work, we investigated the antifungal activity of two de novo designed, antimicrobial peptides {VS2} and {VS3}, incorporating unnatural amino acid α,β-dehydrophenylalanine ({ΔPhe}). We observed that the low-hemolytic peptides could irreversibly inhibit the growth of various Candida species and multidrug resistance strains at {MIC}(80) values ranging from {15.62μM} to {250μM}. Synergy experiments showed that {MIC}(80) of the peptides was drastically reduced in combination with an antifungal drug fluconazole. The dye {PI} uptake assay was used to demonstrate peptide induced cell membrane permeabilization. Intracellular localization of the {FITC}-labeled peptides in Candida albicans was studied by confocal microscopy and {FACS}. Killing kinetics, {PI} uptake assay, and the intracellular presence of {FITC}-peptides suggested that growth inhibition is not solely a consequence of increased membrane permeabilization. We showed that entry of the peptide in Candida cells resulted in accumulation of reactive oxygen species ({ROS}) leading to cell necrosis. Morphological alteration in Candida cells caused by the peptides was visualized by electron microscopy. We propose that de novo designed {VS2} and {VS3} peptides have multiple detrimental effects on target fungi, which ultimately result in cell wall disruption and killing. Therefore, these peptides represent a good template for further design and development as antifungal agents.},
added-at = {2018-12-02T16:09:07.000+0100},
author = {Maurya, Indresh Kumar K. and Pathak, Sarika and Sharma, Monika and Sanwal, Hina and Chaudhary, Preeti and Tupe, Santosh and Deshpande, Mukund and Chauhan, Virander Singh S. and Prasad, Rajendra},
biburl = {https://www.bibsonomy.org/bibtex/284cf0be974ce9017c03bcf1a196c3a2f/karthikraman},
citeulike-article-id = {9479163},
citeulike-linkout-0 = {http://dx.doi.org/10.1016/j.peptides.2011.06.003},
citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/21693143},
citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=21693143},
day = 13,
doi = {10.1016/j.peptides.2011.06.003},
interhash = {6510482d26972b3de15e88f276667787},
intrahash = {84cf0be974ce9017c03bcf1a196c3a2f},
issn = {1873-5169},
journal = {Peptides},
keywords = {ros},
month = jun,
pmid = {21693143},
posted-at = {2011-07-01 05:51:16},
priority = {2},
timestamp = {2018-12-02T16:09:07.000+0100},
title = {Antifungal activity of novel synthetic peptides by accumulation of reactive oxygen species ({ROS}) and disruption of cell wall against Candida albicans.},
url = {http://dx.doi.org/10.1016/j.peptides.2011.06.003},
year = 2011
}