Photo-oxidation-induced inactivation of the selenium-containing protective enzymes thioredoxin reductase and glutathione peroxidase

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Photo-oxidation-induced inactivation of the selenium-containing protective enzymes thioredoxin reductase and glutathione peroxidase. / Suryo Rahmanto, Aldwin; Pattison, David I; Davies, Michael Jonathan.

In: Free Radical Biology & Medicine, Vol. 53, No. 6, 15.09.2012, p. 1308-16.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Suryo Rahmanto, A, Pattison, DI & Davies, MJ 2012, 'Photo-oxidation-induced inactivation of the selenium-containing protective enzymes thioredoxin reductase and glutathione peroxidase', Free Radical Biology & Medicine, vol. 53, no. 6, pp. 1308-16. https://doi.org/10.1016/j.freeradbiomed.2012.07.016

APA

Suryo Rahmanto, A., Pattison, D. I., & Davies, M. J. (2012). Photo-oxidation-induced inactivation of the selenium-containing protective enzymes thioredoxin reductase and glutathione peroxidase. Free Radical Biology & Medicine, 53(6), 1308-16. https://doi.org/10.1016/j.freeradbiomed.2012.07.016

Vancouver

Suryo Rahmanto A, Pattison DI, Davies MJ. Photo-oxidation-induced inactivation of the selenium-containing protective enzymes thioredoxin reductase and glutathione peroxidase. Free Radical Biology & Medicine. 2012 Sep 15;53(6):1308-16. https://doi.org/10.1016/j.freeradbiomed.2012.07.016

Author

Suryo Rahmanto, Aldwin ; Pattison, David I ; Davies, Michael Jonathan. / Photo-oxidation-induced inactivation of the selenium-containing protective enzymes thioredoxin reductase and glutathione peroxidase. In: Free Radical Biology & Medicine. 2012 ; Vol. 53, No. 6. pp. 1308-16.

Bibtex

@article{cc578c7594414596b1166ae383dbf7fb,
title = "Photo-oxidation-induced inactivation of the selenium-containing protective enzymes thioredoxin reductase and glutathione peroxidase",
abstract = "Singlet oxygen ((1)O(2)) is a reactive oxygen species generated during photo-oxidation, inflammation, and via peroxidase-catalyzed reactions (e.g., myeloperoxidase and eosinophil peroxidase). (1)O(2) oxidizes the free amino acids Trp, Tyr, His, Cys, and Met, and those species present on peptides/proteins, with this resulting in modulation of protein structure and function. Impairment of the activity of antioxidant enzymes may be of relevance to the oxidative stress observed in a number of pathologies involving either light exposure or inflammation. In this study, the effects of (1)O(2) on glutathione peroxidase (GPx) and thioredoxin reductase (TrxR) activity, including the mechanisms of their inactivation, were investigated. Exposure of GPx or TrxR, either as purified proteins or in cell lysates, to Rose Bengal and visible light (an established source of (1)O(2)) resulted in significant, photolysis time-dependent reductions in enzyme activity (10-40{\%}, P<0.05). More extensive inhibition (ca. 2-fold) was detected when the reactions were carried out in D(2)O, consistent with the intermediacy of (1)O(2). No additional inhibition was detected after the cessation of photolysis, eliminating a role for photo-products. Methionine, which reacts rapidly with (1)O(2) (k~10(7)M(-1) s(-1))(,) significantly reduced photo-inactivation at large molar excesses, presumably by acting as an alternative target. Reductants (NaBH(4), DTT, GSH, or NADPH) added after the cessation of (1)O(2) formation were unable to reverse enzyme inactivation, consistent with irreversible enzyme oxidation. Formation of nonreducible protein aggregates and/or fragments was detected for both photo-oxidized GPx and TrxR by SDS-PAGE. An oxidant concentration-dependent increase in protein carbonyls was detected with TrxR but not GPx. These studies thus demonstrate that the antioxidant enzymes GPx and TrxR can be irreversibly inactivated by (1)O(2).",
keywords = "Animals, Borohydrides, Cattle, Cell Extracts, Cell Line, Dithiothreitol, Enzyme Stability, Free Radical Scavengers, Glutathione, Glutathione Peroxidase, Hydrogen Peroxide, Methionine, Mice, Oxidants, Photochemical, Oxidation-Reduction, Photolysis, Protein Carbonylation, Rats, Reducing Agents, Rose Bengal, Singlet Oxygen, Thioredoxin-Disulfide Reductase",
author = "{Suryo Rahmanto}, Aldwin and Pattison, {David I} and Davies, {Michael Jonathan}",
note = "Copyright {\circledC} 2012 Elsevier Inc. All rights reserved.",
year = "2012",
month = "9",
day = "15",
doi = "10.1016/j.freeradbiomed.2012.07.016",
language = "English",
volume = "53",
pages = "1308--16",
journal = "Free Radical Biology & Medicine",
issn = "0891-5849",
publisher = "Elsevier",
number = "6",

}

RIS

TY - JOUR

T1 - Photo-oxidation-induced inactivation of the selenium-containing protective enzymes thioredoxin reductase and glutathione peroxidase

AU - Suryo Rahmanto, Aldwin

AU - Pattison, David I

AU - Davies, Michael Jonathan

N1 - Copyright © 2012 Elsevier Inc. All rights reserved.

PY - 2012/9/15

Y1 - 2012/9/15

N2 - Singlet oxygen ((1)O(2)) is a reactive oxygen species generated during photo-oxidation, inflammation, and via peroxidase-catalyzed reactions (e.g., myeloperoxidase and eosinophil peroxidase). (1)O(2) oxidizes the free amino acids Trp, Tyr, His, Cys, and Met, and those species present on peptides/proteins, with this resulting in modulation of protein structure and function. Impairment of the activity of antioxidant enzymes may be of relevance to the oxidative stress observed in a number of pathologies involving either light exposure or inflammation. In this study, the effects of (1)O(2) on glutathione peroxidase (GPx) and thioredoxin reductase (TrxR) activity, including the mechanisms of their inactivation, were investigated. Exposure of GPx or TrxR, either as purified proteins or in cell lysates, to Rose Bengal and visible light (an established source of (1)O(2)) resulted in significant, photolysis time-dependent reductions in enzyme activity (10-40%, P<0.05). More extensive inhibition (ca. 2-fold) was detected when the reactions were carried out in D(2)O, consistent with the intermediacy of (1)O(2). No additional inhibition was detected after the cessation of photolysis, eliminating a role for photo-products. Methionine, which reacts rapidly with (1)O(2) (k~10(7)M(-1) s(-1))(,) significantly reduced photo-inactivation at large molar excesses, presumably by acting as an alternative target. Reductants (NaBH(4), DTT, GSH, or NADPH) added after the cessation of (1)O(2) formation were unable to reverse enzyme inactivation, consistent with irreversible enzyme oxidation. Formation of nonreducible protein aggregates and/or fragments was detected for both photo-oxidized GPx and TrxR by SDS-PAGE. An oxidant concentration-dependent increase in protein carbonyls was detected with TrxR but not GPx. These studies thus demonstrate that the antioxidant enzymes GPx and TrxR can be irreversibly inactivated by (1)O(2).

AB - Singlet oxygen ((1)O(2)) is a reactive oxygen species generated during photo-oxidation, inflammation, and via peroxidase-catalyzed reactions (e.g., myeloperoxidase and eosinophil peroxidase). (1)O(2) oxidizes the free amino acids Trp, Tyr, His, Cys, and Met, and those species present on peptides/proteins, with this resulting in modulation of protein structure and function. Impairment of the activity of antioxidant enzymes may be of relevance to the oxidative stress observed in a number of pathologies involving either light exposure or inflammation. In this study, the effects of (1)O(2) on glutathione peroxidase (GPx) and thioredoxin reductase (TrxR) activity, including the mechanisms of their inactivation, were investigated. Exposure of GPx or TrxR, either as purified proteins or in cell lysates, to Rose Bengal and visible light (an established source of (1)O(2)) resulted in significant, photolysis time-dependent reductions in enzyme activity (10-40%, P<0.05). More extensive inhibition (ca. 2-fold) was detected when the reactions were carried out in D(2)O, consistent with the intermediacy of (1)O(2). No additional inhibition was detected after the cessation of photolysis, eliminating a role for photo-products. Methionine, which reacts rapidly with (1)O(2) (k~10(7)M(-1) s(-1))(,) significantly reduced photo-inactivation at large molar excesses, presumably by acting as an alternative target. Reductants (NaBH(4), DTT, GSH, or NADPH) added after the cessation of (1)O(2) formation were unable to reverse enzyme inactivation, consistent with irreversible enzyme oxidation. Formation of nonreducible protein aggregates and/or fragments was detected for both photo-oxidized GPx and TrxR by SDS-PAGE. An oxidant concentration-dependent increase in protein carbonyls was detected with TrxR but not GPx. These studies thus demonstrate that the antioxidant enzymes GPx and TrxR can be irreversibly inactivated by (1)O(2).

KW - Animals

KW - Borohydrides

KW - Cattle

KW - Cell Extracts

KW - Cell Line

KW - Dithiothreitol

KW - Enzyme Stability

KW - Free Radical Scavengers

KW - Glutathione

KW - Glutathione Peroxidase

KW - Hydrogen Peroxide

KW - Methionine

KW - Mice

KW - Oxidants, Photochemical

KW - Oxidation-Reduction

KW - Photolysis

KW - Protein Carbonylation

KW - Rats

KW - Reducing Agents

KW - Rose Bengal

KW - Singlet Oxygen

KW - Thioredoxin-Disulfide Reductase

U2 - 10.1016/j.freeradbiomed.2012.07.016

DO - 10.1016/j.freeradbiomed.2012.07.016

M3 - Journal article

C2 - 22884457

VL - 53

SP - 1308

EP - 1316

JO - Free Radical Biology & Medicine

JF - Free Radical Biology & Medicine

SN - 0891-5849

IS - 6

ER -

ID: 128974900