Irreversible Thiol Oxidation in Carbonic Anhydrase III: Protection by S-Glutathiolation and Detection in Aging Rats
-
R.J. Mallis
Abstract
Proteins with reactive sulfhydryls are central to many important metabolic reactions and also contribute to a variety of signal transduction systems. In this report, we examine the mechanisms of oxidative damage to the two reactive sulfhydryls of carbonic anhydrase III. Hydrogen peroxide (H2O2), peroxy radicals, or hypochlorous acid (HOCl) produced irreversibly oxidized forms, primarily cysteine sulfinic acid or cysteic acid, of carbonic anhydrase III if glutathione (GSH) was not present. When GSH was approximately equimolar to protein thiols, irreversible oxidation was prevented. H2O2 and peroxyl radicals both generated Sglutathiolated carbonic anhydrase III via partially oxidized protein sulfhydryl intermediates, while HOCl did not cause Sglutathiolation. Thus, oxidative damage from H2O2 or AAPH was prevented by protein Sglutathiolation, while a direct reaction between GSH and oxidant likely prevents HOClmediated protein damage. In cultured rat hepatocytes, carbonic anhydrase III was rapidly Sglutathiolated by menadione. When hepatocyte glutathione was depleted, menadione instead caused irreversible oxidation. We hypothesized that normal depletion of glutathione in aged animals might also lead to an increase in irreversible oxidation. Indeed, both total protein extracts and carbonic anhydrase III contained significantly more cysteine sulfinic acid in older rats compared to young animals. These experiments show that, in the absence of sufficient GSH, oxidation reactions lead to irreversible protein sulfhydryl damage in purified proteins, cellular systems, and whole animals.
Copyright © 2002 by Walter de Gruyter GmbH & Co. KG
Articles in the same Issue
- Highlight: Oxidative Stress
- Highlight: Oxidative Stress
- Peroxiredoxins
- 15-Lipoxygenase-1: A Prooxidant Enzyme
- Reactive Sulfur Species: An Emerging Concept in Oxidative Stress
- The Pathobiochemistry of Nitrogen Dioxide
- Nitric Oxide and Peroxynitrite Interactions with Mitochondria
- Nitric Oxide and Cell Signaling Pathways in Mitochondrial-Dependent Apoptosis
- Fatty Acid Oxidation and Signaling in Apoptosis
- Oxidant-Induced Signaling: Effects of Peroxynitrite and Singlet Oxygen
- The 80th Anniversary of Vitamin E: Beyond Its Antioxidant Properties
- Role of Endogenous Oxidative DNA Damage in Carcinogenesis: What Can We Learn from Repair-Deficient Mice?
- Mammalian Cell Injury Induced by Hypothermia the Emerging Role for Reactive Oxygen Species
- The Chelatable Iron Pool in Living Cells: A Methodically Defined Quantity
- Dietary Flavonoids as Potential Neuroprotectants
- Oxidative Nerve Cell Death in Alzheimers Disease and Stroke: Antioxidants as Neuroprotective Compounds
- A2E and Blue Light in the Retina: The Paradigm of Age-Related Macular Degeneration
- Interactions of Nitric Oxide and Peroxynitrite with Low-Density Lipoprotein
- Non-Antioxidant Properties of Carotenoids
- Proteolytic Response to Oxidative Stress in Mammalian Cells
- Neurospora crassa Catalases, Singlet Oxygen and Cell Differentiation
- The Cellular Hydration State: A Critical Determinant for Cell Death and Survival
- Redox Regulation of Stress Signals: Possible Roles of Dendritic Stellate TRX Producer Cells (DST Cell Types)
- Oxidative Stress, Spermatogenesis and Fertility
- Postprandial Oxidative Stress
- [18O]-Labeled Singlet Oxygen as a Tool for Mechanistic Studies of 8-Oxo-7,8-Dihydroguanine Oxidative Damage: Detection of Spiroiminodihydantoin, Imidazolone and Oxazolone Derivatives
- The Specificity of Lipoxygenase-Catalyzed Lipid Peroxidation and the Effects of Radical- Scavenging Antioxidants
- Human Neutrophils Oxidize Low-Density Lipoprotein by a Hypochlorous Acid- Dependent Mechanism: The Role of Vitamin C
- Perturbation of Lipid Metabolism by Linoleic Acid Hydroperoxide in CaCo-2 Cells
- Irreversible Thiol Oxidation in Carbonic Anhydrase III: Protection by S-Glutathiolation and Detection in Aging Rats
- Green Tea Extract Protects against Early Alcohol-Induced Liver Injury in Rats
- Comparing β-Carotene, Vitamin E and Nitric Oxide as Membrane Antioxidants
- Induction of Thioredoxin Reductase Gene Expression by Peroxynitrite in Human Umbilical Vein Endothelial Cells
- Activation of c-Jun N-Terminal Kinase and Apoptosis in Endothelial Cells Mediated by Endogenous Generation of Hydrogen Peroxide
- Detection of Poly(ADP-Ribose) by Immunocytochemistry: A Sensitive New Method for the Early Identification of UVB- and H2O2-Induced Apoptosis in Keratinocytes
- Efficacy of HOCl Scavenging by Sulfur- Containing Compounds: Antioxidant Activity of Glutathione Disulfide?
- Cysteine Supplementation Prevents Unweighting-Induced Ubiquitination in Association with Redox Regulation in Rat Skeletal Muscle
Articles in the same Issue
- Highlight: Oxidative Stress
- Highlight: Oxidative Stress
- Peroxiredoxins
- 15-Lipoxygenase-1: A Prooxidant Enzyme
- Reactive Sulfur Species: An Emerging Concept in Oxidative Stress
- The Pathobiochemistry of Nitrogen Dioxide
- Nitric Oxide and Peroxynitrite Interactions with Mitochondria
- Nitric Oxide and Cell Signaling Pathways in Mitochondrial-Dependent Apoptosis
- Fatty Acid Oxidation and Signaling in Apoptosis
- Oxidant-Induced Signaling: Effects of Peroxynitrite and Singlet Oxygen
- The 80th Anniversary of Vitamin E: Beyond Its Antioxidant Properties
- Role of Endogenous Oxidative DNA Damage in Carcinogenesis: What Can We Learn from Repair-Deficient Mice?
- Mammalian Cell Injury Induced by Hypothermia the Emerging Role for Reactive Oxygen Species
- The Chelatable Iron Pool in Living Cells: A Methodically Defined Quantity
- Dietary Flavonoids as Potential Neuroprotectants
- Oxidative Nerve Cell Death in Alzheimers Disease and Stroke: Antioxidants as Neuroprotective Compounds
- A2E and Blue Light in the Retina: The Paradigm of Age-Related Macular Degeneration
- Interactions of Nitric Oxide and Peroxynitrite with Low-Density Lipoprotein
- Non-Antioxidant Properties of Carotenoids
- Proteolytic Response to Oxidative Stress in Mammalian Cells
- Neurospora crassa Catalases, Singlet Oxygen and Cell Differentiation
- The Cellular Hydration State: A Critical Determinant for Cell Death and Survival
- Redox Regulation of Stress Signals: Possible Roles of Dendritic Stellate TRX Producer Cells (DST Cell Types)
- Oxidative Stress, Spermatogenesis and Fertility
- Postprandial Oxidative Stress
- [18O]-Labeled Singlet Oxygen as a Tool for Mechanistic Studies of 8-Oxo-7,8-Dihydroguanine Oxidative Damage: Detection of Spiroiminodihydantoin, Imidazolone and Oxazolone Derivatives
- The Specificity of Lipoxygenase-Catalyzed Lipid Peroxidation and the Effects of Radical- Scavenging Antioxidants
- Human Neutrophils Oxidize Low-Density Lipoprotein by a Hypochlorous Acid- Dependent Mechanism: The Role of Vitamin C
- Perturbation of Lipid Metabolism by Linoleic Acid Hydroperoxide in CaCo-2 Cells
- Irreversible Thiol Oxidation in Carbonic Anhydrase III: Protection by S-Glutathiolation and Detection in Aging Rats
- Green Tea Extract Protects against Early Alcohol-Induced Liver Injury in Rats
- Comparing β-Carotene, Vitamin E and Nitric Oxide as Membrane Antioxidants
- Induction of Thioredoxin Reductase Gene Expression by Peroxynitrite in Human Umbilical Vein Endothelial Cells
- Activation of c-Jun N-Terminal Kinase and Apoptosis in Endothelial Cells Mediated by Endogenous Generation of Hydrogen Peroxide
- Detection of Poly(ADP-Ribose) by Immunocytochemistry: A Sensitive New Method for the Early Identification of UVB- and H2O2-Induced Apoptosis in Keratinocytes
- Efficacy of HOCl Scavenging by Sulfur- Containing Compounds: Antioxidant Activity of Glutathione Disulfide?
- Cysteine Supplementation Prevents Unweighting-Induced Ubiquitination in Association with Redox Regulation in Rat Skeletal Muscle
