Tags

Type your tag names separated by a space and hit enter

Thiol and sulfenic acid oxidation of AhpE, the one-cysteine peroxiredoxin from Mycobacterium tuberculosis: kinetics, acidity constants, and conformational dynamics.
Biochemistry. 2009 Oct 13; 48(40):9416-26.B

Abstract

Drug resistance and virulence of Mycobacterium tuberculosis are partially related to the pathogen's antioxidant systems. Peroxide detoxification in this bacterium is achieved by the heme-containing catalase peroxidase and different two-cysteine peroxiredoxins. M. tuberculosis genome also codifies for a putative one-cysteine peroxiredoxin, alkyl hydroperoxide reductase E (MtAhpE). Its expression was previously demonstrated at a transcriptional level, and the crystallographic structure of the recombinant protein was resolved under reduced and oxidized states. Herein, we report that the conformation of MtAhpE changed depending on its single cysteine redox state, as reflected by different tryptophan fluorescence properties and changes in quaternary structure. Dynamics of fluorescence changes, complemented by competition kinetic assays, were used to perform protein functional studies. MtAhpE reduced peroxynitrite 2 orders of magnitude faster than hydrogen peroxide (1.9 x 10(7) M(-1) s(-1) vs 8.2 x 10(4) M(-1) s(-1) at pH 7.4 and 25 degrees C, respectively). The latter also caused cysteine overoxidation to sulfinic acid, but at much slower rate constant (40 M(-1) s(-1)). The pK(a) of the thiol in the reduced enzyme was 5.2, more than one unit lower than that of the sulfenic acid in the oxidized enzyme. The pH profile of hydrogen peroxide-mediated thiol and sulfenic acid oxidations indicated thiolate and sulfenate as the reacting species. The formation of sulfenic acid as well as the catalytic peroxidase activity of MtAhpE was demonstrated using the artificial reducing substrate thionitrobenzoate. Taken together, our results indicate that MtAhpE is a relevant component in the antioxidant repertoire of M. tuberculosis probably involved in peroxide and specially peroxynitrite detoxification.

Authors+Show Affiliations

Departamento de Bioqumica, Facultad de Medicina, Center for Free Radical and Biomedical Research, Universidad de la República, Montevideo, Uruguay.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19737009

Citation

Hugo, Martín, et al. "Thiol and Sulfenic Acid Oxidation of AhpE, the One-cysteine Peroxiredoxin From Mycobacterium Tuberculosis: Kinetics, Acidity Constants, and Conformational Dynamics." Biochemistry, vol. 48, no. 40, 2009, pp. 9416-26.
Hugo M, Turell L, Manta B, et al. Thiol and sulfenic acid oxidation of AhpE, the one-cysteine peroxiredoxin from Mycobacterium tuberculosis: kinetics, acidity constants, and conformational dynamics. Biochemistry. 2009;48(40):9416-26.
Hugo, M., Turell, L., Manta, B., Botti, H., Monteiro, G., Netto, L. E., Alvarez, B., Radi, R., & Trujillo, M. (2009). Thiol and sulfenic acid oxidation of AhpE, the one-cysteine peroxiredoxin from Mycobacterium tuberculosis: kinetics, acidity constants, and conformational dynamics. Biochemistry, 48(40), 9416-26. https://doi.org/10.1021/bi901221s
Hugo M, et al. Thiol and Sulfenic Acid Oxidation of AhpE, the One-cysteine Peroxiredoxin From Mycobacterium Tuberculosis: Kinetics, Acidity Constants, and Conformational Dynamics. Biochemistry. 2009 Oct 13;48(40):9416-26. PubMed PMID: 19737009.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Thiol and sulfenic acid oxidation of AhpE, the one-cysteine peroxiredoxin from Mycobacterium tuberculosis: kinetics, acidity constants, and conformational dynamics. AU - Hugo,Martín, AU - Turell,Lucía, AU - Manta,Bruno, AU - Botti,Horacio, AU - Monteiro,Gisele, AU - Netto,Luis E S, AU - Alvarez,Beatriz, AU - Radi,Rafael, AU - Trujillo,Madia, PY - 2009/9/10/entrez PY - 2009/9/10/pubmed PY - 2009/12/16/medline SP - 9416 EP - 26 JF - Biochemistry JO - Biochemistry VL - 48 IS - 40 N2 - Drug resistance and virulence of Mycobacterium tuberculosis are partially related to the pathogen's antioxidant systems. Peroxide detoxification in this bacterium is achieved by the heme-containing catalase peroxidase and different two-cysteine peroxiredoxins. M. tuberculosis genome also codifies for a putative one-cysteine peroxiredoxin, alkyl hydroperoxide reductase E (MtAhpE). Its expression was previously demonstrated at a transcriptional level, and the crystallographic structure of the recombinant protein was resolved under reduced and oxidized states. Herein, we report that the conformation of MtAhpE changed depending on its single cysteine redox state, as reflected by different tryptophan fluorescence properties and changes in quaternary structure. Dynamics of fluorescence changes, complemented by competition kinetic assays, were used to perform protein functional studies. MtAhpE reduced peroxynitrite 2 orders of magnitude faster than hydrogen peroxide (1.9 x 10(7) M(-1) s(-1) vs 8.2 x 10(4) M(-1) s(-1) at pH 7.4 and 25 degrees C, respectively). The latter also caused cysteine overoxidation to sulfinic acid, but at much slower rate constant (40 M(-1) s(-1)). The pK(a) of the thiol in the reduced enzyme was 5.2, more than one unit lower than that of the sulfenic acid in the oxidized enzyme. The pH profile of hydrogen peroxide-mediated thiol and sulfenic acid oxidations indicated thiolate and sulfenate as the reacting species. The formation of sulfenic acid as well as the catalytic peroxidase activity of MtAhpE was demonstrated using the artificial reducing substrate thionitrobenzoate. Taken together, our results indicate that MtAhpE is a relevant component in the antioxidant repertoire of M. tuberculosis probably involved in peroxide and specially peroxynitrite detoxification. SN - 1520-4995 UR - https://www.unboundmedicine.com/medline/citation/19737009/Thiol_and_sulfenic_acid_oxidation_of_AhpE_the_one_cysteine_peroxiredoxin_from_Mycobacterium_tuberculosis:_kinetics_acidity_constants_and_conformational_dynamics_ L2 - https://dx.doi.org/10.1021/bi901221s DB - PRIME DP - Unbound Medicine ER -