Please use this identifier to cite or link to this item: http://hdl.handle.net/20.500.12857/116941
Title: The antibacterial prodrug activator Rv2466c is a mycothiol-dependent reductase in the oxidative stress response of Mycobacterium tuberculosis
Authors: Rosado, Leonardo Astolfi
Wahni, Khadija
Degiacomi, Giulia 
Pedre, Brandán
Young, David
de la Rubia, Alfonso G
Boldrin, Francesca 
Martens, Edo
Marcos-Pascual, Laura
Sancho-Vaello, Enea
Albesa-Jové, David
Provvedi, Roberta 
Martin, Charlotte
Makarov, Vadim
Versées, Wim
Verniest, Guido
Guerin, Marcelo E
Mateos, Luis M
Manganelli, Riccardo 
Messens, Joris
Keywords: Mycobacterium tuberculosis;enzyme catalysis;oxidation-reduction (redox);oxidative stress;oxidoreductase;phylogenetics;thiol–disulfide exchange
Keywords Plus: ESCHERICHIA-COLI THIOREDOXIN;METHIONINE SULFOXIDE REDUCTASE;CORYNEBACTERIUM-GLUTAMICUM;THIOL REDOX;MAXIMUM-ENTROPY;GLUTAREDOXIN;PROTEIN;DISULFIDE;MECHANISM;RESISTANCE
Mesh headings: Anti-Bacterial Agents;Bacterial Proteins;Models, Molecular;Mycobacterium tuberculosis;Oxidative Stress;Prodrugs;Protein Disulfide-Isomerases;Pyrimidines
Secondary Mesh headings: Activation, Metabolic;Biocatalysis;Catalytic Domain;Crystallography, X-Ray;Cysteine;Disk Diffusion Antimicrobial Tests;Drugs, Investigational;Gene Deletion;Molecular Conformation;Molecular Docking Simulation;Oxidation-Reduction;Phylogeny;Protein Conformation;Recombinant Proteins;Substrate Specificity
Issue Date: 2017
Publisher: AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Journal: The Journal of biological chemistry 
Abstract: 
The Mycobacterium tuberculosis rv2466c gene encodes an oxidoreductase enzyme annotated as DsbA. It has a CPWC active-site motif embedded within its thioredoxin fold domain and mediates the activation of the prodrug TP053, a thienopyrimidine derivative that kills both replicating and nonreplicating bacilli. However, its mode of action and actual enzymatic function in M. tuberculosis have remained enigmatic. In this study, we report that Rv2466c is essential for bacterial survival under H2O2 stress. Further, we discovered that Rv2466c lacks oxidase activity; rather, it receives electrons through the mycothiol/mycothione reductase/NADPH pathway to activate TP053, preferentially via a dithiol-disulfide mechanism. We also found that Rv2466c uses a monothiol-disulfide exchange mechanism to reduce S-mycothiolated mixed disulfides and intramolecular disulfides. Genetic, phylogenetic, bioinformatics, structural, and biochemical analyses revealed that Rv2466c is a novel mycothiol-dependent reductase, which represents a mycoredoxin cluster of enzymes within the DsbA family different from the glutaredoxin cluster to which mycoredoxin-1 (Mrx1 or Rv3198A) belongs. To validate this DsbA-mycoredoxin cluster, we also characterized a homologous enzyme of Corynebacterium glutamicum (NCgl2339) and observed that it demycothiolates and reduces a mycothiol arsenate adduct with kinetic properties different from those of Mrx1. In conclusion, our work has uncovered a DsbA-like mycoredoxin that promotes mycobacterial resistance to oxidative stress and reacts with free mycothiol and mycothiolated targets. The characterization of the DsbA-like mycoredoxin cluster reported here now paves the way for correctly classifying similar enzymes from other organisms.
URI: http://hdl.handle.net/20.500.12857/116941
ISSN: 00219258
DOI: 10.1074/jbc.M117.797837
Appears in Collections:Articles

Show full item record

PubMed Central
Citations 50

9
Last Week
0
Last month
0
checked on Dec 5, 2021

SCOPUSTM   
Citations

12
checked on Aug 31, 2020

WEB OF SCIENCETM
Citations

15
checked on Dec 2, 2021

Page view(s) 5

2
checked on Dec 6, 2021

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.