title: Proteomic responses to a methyl viologen-induced oxidative stress in the wild type and FerB mutant strains of Paracoccus denitrificans.
abstract: FerB is a cytoplasmic flavoprotein from the soil bacterium Paracoccus denitrificans with a putative role in defense against oxidative stress. To further explore this hypothesis, we compared protein variations upon methyl viologen treatment in wild-type and FerB mutant strains by a quantitative proteomic analysis based on iTRAQ-3DLC-MS/MS analysis. The proteins showing the most prominent increase in abundance were assigned to carbon fixation and sulfur assimilatory pathways. By employing these proteins as indirect markers, oxidative stress was found to be 15% less severe in the wild-type than in the FerB-deficient mutant cells. Oxidative stress altered the levels of proteins whose expression is dependent on the transcriptional factor FnrP. The observed down-regulation of the fnrP regulon members, most notably that of nitrous oxide reductase, was tentatively explained by an oxidative degradation of the [4Fe-4S] center of FnrP leading to a protein form which no longer activates transcription. While the level of FerB remained relatively constant, two proteins homologous to FerB accumulated during oxidative stress. When their genes were expressed in Escherichia coli, neither of the protein products contained a bound flavin, whereas they both had a high activity of flavin reductase, one preferentially utilizing NADH and the other NADPH.
authors.affiliation: Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic.
authors.forename: Vendula
authors.initials: V
authors.lastname: Pernikářová
authors.affiliation: Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic.
authors.forename: Vojtěch
authors.initials: V
authors.lastname: Sedláček
authors.affiliation: Central European Institute for Technology, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic; National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic.
authors.forename: David
authors.initials: D
authors.lastname: Potěšil
authors.affiliation: Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic; Regional Centre for Applied Molecular Oncology, Masaryk Memorial Cancer Institute, Žlutý kopec 7, 65653 Brno, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Procházková
authors.affiliation: Central European Institute for Technology, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic; National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic.
authors.forename: Zbyněk
authors.initials: Z
authors.lastname: Zdráhal
authors.affiliation: Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic. Electronic address: bouchal@chemi.muni.cz.
authors.forename: Pavel
authors.initials: P
authors.lastname: Bouchal
authors.affiliation: Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic. Electronic address: ikucera@chemi.muni.cz.
authors.forename: Igor
authors.initials: I
authors.lastname: Kučera
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D005420:Flavoproteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects* / Q000235:genetics; D009154:Mutation; D018384:Oxidative Stress / Q000187:drug effects* / Q000235:genetics; D010231:Paracoccus denitrificans / Q000235:genetics / Q000378:metabolism*; D010269:Paraquat / Q000494:pharmacology*; D040901:Proteomics
chemical_list: D001426:Bacterial Proteins; D005420:Flavoproteins; D010269:Paraquat
keywords: Flavin reductase; Flavoprotein; Methyl viologen; Microbial proteomics; Oxidative stress; Paracoccus denitrificans