PrxQ B from Mycobacterium tuberculosis is a monomeric, thioredoxin-dependent and highly efficient fatty acid hydroperoxide reductase
Mycobacterium tuberculosis (M. tuberculosis) is the intracellular bacterium responsible for tuberculosis disease (TD). Inside the phagosomes of activated macrophages, M. tuberculosis is exposed to cytotoxic hydroperoxides such as hydrogen peroxide, fatty acid hydroperoxides and peroxynitrite. Thus,...
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todo:paper_08915849_v101_n_p249_Reyes2023-10-03T15:41:20Z PrxQ B from Mycobacterium tuberculosis is a monomeric, thioredoxin-dependent and highly efficient fatty acid hydroperoxide reductase Reyes, A.M. Vazquez, D.S. Zeida, A. Hugo, M. Piñeyro, M.D. De Armas, M.I. Estrin, D. Radi, R. Santos, J. Trujillo, M. Fatty acid hydroperoxides Mycobacterium tuberculosis Peroxidatic and resolving cysteine Peroxiredoxin Peroxynitrite Thiol-dependent peroxidase Thioredoxin deoxycholate sodium oxidoreductase peroxiredoxin Q B peroxynitrite reducing agent thioredoxin unclassified drug aldehyde dehydrogenase bacterial protein fatty acid fatty acid reductase hydrogen peroxide peroxiredoxin protein binding recombinant protein thioredoxin Article bacterial growth catalysis circular dichroism conformational transition enzyme activity enzyme conformation hydrodynamics hydrogen bond hydrophobicity molecular dynamics Mycobacterium tuberculosis nonhuman observed rate constant oxidation oxidation reduction state priority journal protein expression protein secondary structure protein structure protein tertiary structure reduction sequence homology steady state alpha helix beta sheet binding site chemistry enzyme specificity enzymology Escherichia coli gene expression gene vector genetics kinetics metabolism molecular cloning Mycobacterium tuberculosis oxidation reduction reaction protein domain protein motif Aldehyde Oxidoreductases Amino Acid Motifs Bacterial Proteins Binding Sites Cloning, Molecular Escherichia coli Fatty Acids Gene Expression Genetic Vectors Hydrogen Peroxide Kinetics Molecular Dynamics Simulation Mycobacterium tuberculosis Oxidation-Reduction Peroxiredoxins Protein Binding Protein Conformation, alpha-Helical Protein Conformation, beta-Strand Protein Interaction Domains and Motifs Recombinant Proteins Substrate Specificity Thioredoxins Mycobacterium tuberculosis (M. tuberculosis) is the intracellular bacterium responsible for tuberculosis disease (TD). Inside the phagosomes of activated macrophages, M. tuberculosis is exposed to cytotoxic hydroperoxides such as hydrogen peroxide, fatty acid hydroperoxides and peroxynitrite. Thus, the characterization of the bacterial antioxidant systems could facilitate novel drug developments. In this work, we characterized the product of the gene Rv1608c from M. tuberculosis, which according to sequence homology had been annotated as a putative peroxiredoxin of the peroxiredoxin Q subfamily (PrxQ B from M. tuberculosis or MtPrxQ B). The protein has been reported to be essential for M. tuberculosis growth in cholesterol-rich medium. We demonstrated the M. tuberculosis thioredoxin B/C-dependent peroxidase activity of MtPrxQ B, which acted as a two-cysteine peroxiredoxin that could function, although less efficiently, using a one-cysteine mechanism. Through steady-state and competition kinetic analysis, we proved that the net forward rate constant of MtPrxQ B reaction was 3 orders of magnitude faster for fatty acid hydroperoxides than for hydrogen peroxide (3×106 vs 6×103 M− 1 s− 1, respectively), while the rate constant of peroxynitrite reduction was (0.6−1.4) ×106 M− 1 s− 1 at pH 7.4. The enzyme lacked activity towards cholesterol hydroperoxides solubilized in sodium deoxycholate. Both thioredoxin B and C rapidly reduced the oxidized form of MtPrxQ B, with rates constants of 0.5×106 and 1×106 M− 1 s− 1, respectively. Our data indicated that MtPrxQ B is monomeric in solution both under reduced and oxidized states. In spite of the similar hydrodynamic behavior the reduced and oxidized forms of the protein showed important structural differences that were reflected in the protein circular dichroism spectra. © 2016 Elsevier Inc. Fil:Estrin, D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Santos, J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_08915849_v101_n_p249_Reyes |
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Universidad de Buenos Aires |
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I-28 |
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R-134 |
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Fatty acid hydroperoxides Mycobacterium tuberculosis Peroxidatic and resolving cysteine Peroxiredoxin Peroxynitrite Thiol-dependent peroxidase Thioredoxin deoxycholate sodium oxidoreductase peroxiredoxin Q B peroxynitrite reducing agent thioredoxin unclassified drug aldehyde dehydrogenase bacterial protein fatty acid fatty acid reductase hydrogen peroxide peroxiredoxin protein binding recombinant protein thioredoxin Article bacterial growth catalysis circular dichroism conformational transition enzyme activity enzyme conformation hydrodynamics hydrogen bond hydrophobicity molecular dynamics Mycobacterium tuberculosis nonhuman observed rate constant oxidation oxidation reduction state priority journal protein expression protein secondary structure protein structure protein tertiary structure reduction sequence homology steady state alpha helix beta sheet binding site chemistry enzyme specificity enzymology Escherichia coli gene expression gene vector genetics kinetics metabolism molecular cloning Mycobacterium tuberculosis oxidation reduction reaction protein domain protein motif Aldehyde Oxidoreductases Amino Acid Motifs Bacterial Proteins Binding Sites Cloning, Molecular Escherichia coli Fatty Acids Gene Expression Genetic Vectors Hydrogen Peroxide Kinetics Molecular Dynamics Simulation Mycobacterium tuberculosis Oxidation-Reduction Peroxiredoxins Protein Binding Protein Conformation, alpha-Helical Protein Conformation, beta-Strand Protein Interaction Domains and Motifs Recombinant Proteins Substrate Specificity Thioredoxins |
| spellingShingle |
Fatty acid hydroperoxides Mycobacterium tuberculosis Peroxidatic and resolving cysteine Peroxiredoxin Peroxynitrite Thiol-dependent peroxidase Thioredoxin deoxycholate sodium oxidoreductase peroxiredoxin Q B peroxynitrite reducing agent thioredoxin unclassified drug aldehyde dehydrogenase bacterial protein fatty acid fatty acid reductase hydrogen peroxide peroxiredoxin protein binding recombinant protein thioredoxin Article bacterial growth catalysis circular dichroism conformational transition enzyme activity enzyme conformation hydrodynamics hydrogen bond hydrophobicity molecular dynamics Mycobacterium tuberculosis nonhuman observed rate constant oxidation oxidation reduction state priority journal protein expression protein secondary structure protein structure protein tertiary structure reduction sequence homology steady state alpha helix beta sheet binding site chemistry enzyme specificity enzymology Escherichia coli gene expression gene vector genetics kinetics metabolism molecular cloning Mycobacterium tuberculosis oxidation reduction reaction protein domain protein motif Aldehyde Oxidoreductases Amino Acid Motifs Bacterial Proteins Binding Sites Cloning, Molecular Escherichia coli Fatty Acids Gene Expression Genetic Vectors Hydrogen Peroxide Kinetics Molecular Dynamics Simulation Mycobacterium tuberculosis Oxidation-Reduction Peroxiredoxins Protein Binding Protein Conformation, alpha-Helical Protein Conformation, beta-Strand Protein Interaction Domains and Motifs Recombinant Proteins Substrate Specificity Thioredoxins Reyes, A.M. Vazquez, D.S. Zeida, A. Hugo, M. Piñeyro, M.D. De Armas, M.I. Estrin, D. Radi, R. Santos, J. Trujillo, M. PrxQ B from Mycobacterium tuberculosis is a monomeric, thioredoxin-dependent and highly efficient fatty acid hydroperoxide reductase |
| topic_facet |
Fatty acid hydroperoxides Mycobacterium tuberculosis Peroxidatic and resolving cysteine Peroxiredoxin Peroxynitrite Thiol-dependent peroxidase Thioredoxin deoxycholate sodium oxidoreductase peroxiredoxin Q B peroxynitrite reducing agent thioredoxin unclassified drug aldehyde dehydrogenase bacterial protein fatty acid fatty acid reductase hydrogen peroxide peroxiredoxin protein binding recombinant protein thioredoxin Article bacterial growth catalysis circular dichroism conformational transition enzyme activity enzyme conformation hydrodynamics hydrogen bond hydrophobicity molecular dynamics Mycobacterium tuberculosis nonhuman observed rate constant oxidation oxidation reduction state priority journal protein expression protein secondary structure protein structure protein tertiary structure reduction sequence homology steady state alpha helix beta sheet binding site chemistry enzyme specificity enzymology Escherichia coli gene expression gene vector genetics kinetics metabolism molecular cloning Mycobacterium tuberculosis oxidation reduction reaction protein domain protein motif Aldehyde Oxidoreductases Amino Acid Motifs Bacterial Proteins Binding Sites Cloning, Molecular Escherichia coli Fatty Acids Gene Expression Genetic Vectors Hydrogen Peroxide Kinetics Molecular Dynamics Simulation Mycobacterium tuberculosis Oxidation-Reduction Peroxiredoxins Protein Binding Protein Conformation, alpha-Helical Protein Conformation, beta-Strand Protein Interaction Domains and Motifs Recombinant Proteins Substrate Specificity Thioredoxins |
| description |
Mycobacterium tuberculosis (M. tuberculosis) is the intracellular bacterium responsible for tuberculosis disease (TD). Inside the phagosomes of activated macrophages, M. tuberculosis is exposed to cytotoxic hydroperoxides such as hydrogen peroxide, fatty acid hydroperoxides and peroxynitrite. Thus, the characterization of the bacterial antioxidant systems could facilitate novel drug developments. In this work, we characterized the product of the gene Rv1608c from M. tuberculosis, which according to sequence homology had been annotated as a putative peroxiredoxin of the peroxiredoxin Q subfamily (PrxQ B from M. tuberculosis or MtPrxQ B). The protein has been reported to be essential for M. tuberculosis growth in cholesterol-rich medium. We demonstrated the M. tuberculosis thioredoxin B/C-dependent peroxidase activity of MtPrxQ B, which acted as a two-cysteine peroxiredoxin that could function, although less efficiently, using a one-cysteine mechanism. Through steady-state and competition kinetic analysis, we proved that the net forward rate constant of MtPrxQ B reaction was 3 orders of magnitude faster for fatty acid hydroperoxides than for hydrogen peroxide (3×106 vs 6×103 M− 1 s− 1, respectively), while the rate constant of peroxynitrite reduction was (0.6−1.4) ×106 M− 1 s− 1 at pH 7.4. The enzyme lacked activity towards cholesterol hydroperoxides solubilized in sodium deoxycholate. Both thioredoxin B and C rapidly reduced the oxidized form of MtPrxQ B, with rates constants of 0.5×106 and 1×106 M− 1 s− 1, respectively. Our data indicated that MtPrxQ B is monomeric in solution both under reduced and oxidized states. In spite of the similar hydrodynamic behavior the reduced and oxidized forms of the protein showed important structural differences that were reflected in the protein circular dichroism spectra. © 2016 Elsevier Inc. |
| format |
JOUR |
| author |
Reyes, A.M. Vazquez, D.S. Zeida, A. Hugo, M. Piñeyro, M.D. De Armas, M.I. Estrin, D. Radi, R. Santos, J. Trujillo, M. |
| author_facet |
Reyes, A.M. Vazquez, D.S. Zeida, A. Hugo, M. Piñeyro, M.D. De Armas, M.I. Estrin, D. Radi, R. Santos, J. Trujillo, M. |
| author_sort |
Reyes, A.M. |
| title |
PrxQ B from Mycobacterium tuberculosis is a monomeric, thioredoxin-dependent and highly efficient fatty acid hydroperoxide reductase |
| title_short |
PrxQ B from Mycobacterium tuberculosis is a monomeric, thioredoxin-dependent and highly efficient fatty acid hydroperoxide reductase |
| title_full |
PrxQ B from Mycobacterium tuberculosis is a monomeric, thioredoxin-dependent and highly efficient fatty acid hydroperoxide reductase |
| title_fullStr |
PrxQ B from Mycobacterium tuberculosis is a monomeric, thioredoxin-dependent and highly efficient fatty acid hydroperoxide reductase |
| title_full_unstemmed |
PrxQ B from Mycobacterium tuberculosis is a monomeric, thioredoxin-dependent and highly efficient fatty acid hydroperoxide reductase |
| title_sort |
prxq b from mycobacterium tuberculosis is a monomeric, thioredoxin-dependent and highly efficient fatty acid hydroperoxide reductase |
| url |
http://hdl.handle.net/20.500.12110/paper_08915849_v101_n_p249_Reyes |
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