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title Protein profiles of Escherichia coli and Staphylococcus warneri are altered by photosensitization with cationic porphyrins
authors Alves, E; Esteves, AC; Correia, A; Cunha, A; Faustino, MAF; Neves, MGPMS; Almeida, A
author full name Alves, Eliana; Esteves, Ana Cristina; Correia, Antonio; Cunha, Angela; Faustino, Maria A. F.; Neves, Maria G. P. M. S.; Almeida, Adelaide
title Protein profiles of Escherichia coli and Staphylococcus warneri are altered by photosensitization with cationic porphyrins
nationality internacional
language English
document type Article
abstract Oxidative stress induced by photodynamic treatment of microbial cells causes irreversible damages to vital cellular components such as proteins. Photodynamic inactivation (PDI) of bacteria, a promising therapeutic approach for the treatment of superficial and localized skin and oral infections, can be achieved by exciting a photosensitizing agent with visible light in an oxygenated environment. Although some studies have addressed the oxidative alterations of PDI in bacterial proteins, the present study is the first to compare the electrophoretic profiles of proteins of Gram-positive and Gram-negative bacteria, having two structurally different porphyrins, with different kinetics of photoinactivation. The cationic porphyrins 5,10,15-tris(1-methylpyridinium-4-yl)-20-(pentafluorophenyl)porphyrin tri-iodide (Tri-Py+-Me-PF) and 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)porphyrin tetra-iodide (Tetra-Py+-Me) were used to photosensitize Escherichia coli and Staphylococcus warneri upon white light irradiation at an irradiance of 4.0 mW cm(-2). After different photosensitization periods, proteins were extracted from bacteria and analyzed using one-dimensional SDS-PAGE. Apparent molecular weights and band intensities were determined after an irradiation period corresponding to a reduction of 4 log(10) in cell viability. After photodynamic treatment, there was a general loss of bacterial proteins, assigned to large-scale protein degradation. Protein loss was more pronounced after PDI with Tri-Py+-Me-PF in both bacteria. There was also an increase in the concentration of some proteins as well as an increase in the molecular weight of other proteins. We show that proteins of E. coli and S. warneri are important targets of PDI. Although there is an attempt of cellular response to the PDI-induced damage by overexpression of a limited number of proteins, the damage is lethal. Our results show that changes occurring in the protein pattern during photodynamic treatment are different with the two photosensitizers, which helps to explain the different inactivation kinetics of the two bacteria. SDS-PAGE is a rational approach to assign the type of cellular response to stress that is being induced in the cells.
author address [Alves, Eliana; Esteves, Ana Cristina; Correia, Antonio; Cunha, Angela; Almeida, Adelaide] Univ Aveiro, Dept Biol, P-3810193 Aveiro, Portugal; [Alves, Eliana; Esteves, Ana Cristina; Correia, Antonio; Cunha, Angela; Almeida, Adelaide] Univ Aveiro, Ctr Environm & Marine Studies CESAM, P-3810193 Aveiro, Portugal; [Faustino, Maria A. F.; Neves, Maria G. P. M. S.] Univ Aveiro, Dept Chem, P-3810193 Aveiro, Portugal; [Faustino, Maria A. F.; Neves, Maria G. P. M. S.] Univ Aveiro, Organ Chem Res Unit QOPNA, P-3810193 Aveiro, Portugal
reprint address Neves, MGPMS (reprint author), Univ Aveiro, Dept Chem, P-3810193 Aveiro, Portugal.
e-mail address aalmeida@ua.pt
researcherid number Correia, Antonio/B-1593-2008; Almeida, Adelaide/D-9899-2011; Esteves, Ana Cristina/B-2939-2008; Cunha, Angela/E-9537-2011; Alves, Eliana/K-3158-2013; CESAM, UA/M-3762-2015; Faustino, Maria Amparo/J-5787-2012
orcid number Alves, Eliana/0000-0001-5396-6536; Correia, Antonio/0000-0002-5115-1429; Almeida, Adelaide/0000-0002-8422-8664; Esteves, Ana Cristina/0000-0003-2239-2976; Cunha, Angela/0000-0002-9118-3521; Faustino, Maria Amparo/0000-0003-4423-3802
funding agency and grant number University of Aveiro; Fundacao para a Ciencia e a Tecnologia (FCT, Portugal); European Union; QREN; COMPETE; FEDER [Pest-C/MAR/LA0017/2013, PEst-C/QUI/UI0062/2013, FCOMP-01-0124-FEDER-037296]; FCT [BD/41806/2007, BPD/38008/2007]
funding text The authors are thankful to the University of Aveiro, Fundacao para a Ciencia e a Tecnologia (FCT, Portugal), European Union, QREN, COMPETE and FEDER for funding the Centre for Environmental and Marine Studies (CESAM) unit (project Pest-C/MAR/LA0017/2013) and the QOPNA research unit (project PEst-C/QUI/UI0062/2013, FCOMP-01-0124-FEDER-037296). Eliana Alves (BD/41806/2007) and Ana Cristina Esteves (BPD/38008/2007) are grateful to FCT for their grants.
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cited reference count 57
publisher city CAMBRIDGE
issn 1474-905X
29-character source abbreviation PHOTOCH PHOTOBIO SCI
iso source abbreviation Photochem. Photobiol. Sci.
year published 2015
volume 14
issue 6
beginning page 1169
ending page 1178
digital object identifier (doi) 10.1039/c4pp00194j
page count 10
web of science category Biochemistry & Molecular Biology; Biophysics; Chemistry, Physical
subject category Biochemistry & Molecular Biology; Biophysics; Chemistry
document delivery number CJ8FD
unique article identifier WOS:000355734700011