The Biological Buffer Bicarbonate/CO2 Potentiates H2O2-Mediated Inactivation of Protein Tyrosine Phosphatases

Zhou, Haiying; Singh, Harkewal; Parsons, Zachary D.; Lewis, Sally; Bhattacharya, Sanjib; Seiner, Derrick R.; Labutti, Jason; Reilly, Thomas J.; Tanner, John J.; Gates, Kent S.

doi:10.1021/ja2077137

Cited by 70 publications

(95 citation statements)

References 33 publications

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“…One possibility is that enzymes with hypersensitive cysteine residues are out there, but workers have not yet chanced upon them. Second, bicarbonate accelerates H 2 O 2 -driven thiol oxidation 122, 123 , raising the prospect that in CO 2 -rich environments, disulfide formation might be relevant. A third possibility is that these redoxins help restore the coordinating cysteine ligand that can be oxidized by the ferryl radical in the active sites of mononuclear iron enzymes (Fig.…”

Section: Damage Caused By O2− and H2o2mentioning

confidence: 99%

The molecular mechanisms and physiological consequences of oxidative stress: lessons from a model bacterium

2013

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Oxic environments are hazardous. Molecular oxygen adventitiously abstracts electrons from many redox enzymes, continuously forming intracellular superoxide and hydrogen peroxide. These species can destroy the activities of metalloenzymes and the integrity of DNA, which forces organisms to protect themselves with scavenging enzymes and repair systems. Nevertheless, elevated levels of oxidants quickly poison bacteria, and both microbial competitors and hostile eukaryotic hosts exploit this vulnerability by assaulting them with peroxides or superoxide-forming antibiotics. In response, bacteria activate elegant adaptive strategies. In this Review, I summarize our current knowledge of oxidative stress in Escherichia coli, the model organism for which our understanding of damage and defence is most well-developed.

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Section: Damage Caused By O2− and H2o2mentioning

confidence: 99%

The molecular mechanisms and physiological consequences of oxidative stress: lessons from a model bacterium

2013

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“…For example, H 2 O 2 can irreversibly inactivate the human brain type of creatine kinase via a two-stage process, via modification of CYS283, the catalytic residue [91]. Tyrosine phosphatases [92], porcine kidney betaine aldehyde dehydrogenase [93], and D-amino acid oxidase [90,94] are inactivated in a similar way.…”

Section: Inactivation Of Enzymes By Modification With Hydrogen Peroxidementioning

confidence: 99%

Hydrogen Peroxide in Biocatalysis. A Dangerous Liaison

Hernández

Berenguer-Murcia²,

Rodrigues³

et al. 2012

COC

140

107

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Hydrogen peroxide is a substrate or side-product in many enzyme-catalyzed reactions. For example, it is a side-product of oxidases, resulting from the re-oxidation of FAD with molecular oxygen, and it is a substrate for peroxidases and other enzymes. However, hydrogen peroxide is able to chemically modify the peptide core of the enzymes it interacts with, and also to produce the oxidation of some cofactors and prostetic groups (e.g., the hemo group). Thus, the development of strategies that may permit to increase the stability of enzymes in the presence of this deleterious reagent is an interesting target. This enhancement in enzyme stability has been attempted following almost all available strategies: site-directed mutagenesis (eliminating the most reactive moieties), medium engineering (using stabilizers), immobilization and chemical modification (trying to generate hydrophobic environments surrounding the enzyme, to confer higher rigidity to the protein or to generate oxidation-resistant groups), or the use of systems capable of decomposing hydrogen peroxide under very mild conditions. If hydrogen peroxide is just a side-product, its immediate removal has been reported to be the best solution. In some cases, when hydrogen peroxide is the substrate and its decomposition is not a sensible solution, researchers coupled one enzyme generating hydrogen peroxide "in situ" to the target enzyme resulting in a continuous supply of this reagent at low concentrations thus preventing enzyme inactivation.This review will focus on the general role of hydrogen peroxide in biocatalysis, the main mechanisms of enzyme inactivation produced by this reactive and the different strategies used to prevent enzyme inactivation caused by this "dangerous liaison". Outlook

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“…13 VHR was cloned into an expression vector containing a TEV cleavable 6x-His tag via ligation independent cloning (LIC) techniques. The cDNA for VHR (HsCD00000599) and the LIC vector, pMCSG28 (EvNO00340964) were purchased from DNAsu.…”

Section: Methodsmentioning

confidence: 99%

An immunochemical approach to detect oxidized protein tyrosine phosphatases using a selective C-nucleophile tag

García

Carroll

2016

Mol. BioSyst.

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Protein tyrosine phosphatases are crucial regulators of signal transduction and function as antagonists towards protein tyrosine kinases to control reversible tyrosine phosphorylation, thereby regulating fundamental physiological processes. Growing evidence has supported the notion that reversible oxidative inactivation of the catalytic cysteine residue in protein tyrosine phosphatases serves as an oxidative post-translational modification that regulates its activity to influence downstream signaling by promoting phosphorylation and induction of the signaling cascade. The oxidation of cysteine to the sulfenic acid is often transient and difficult to detect, thus making it problematic in understanding the role that this oxidative post-translational modification plays in redox-biology and pathogenesis. Several methods to detect cysteine oxidation in biological systems have been developed, though targeted approached to directly detect oxidized phosphatases are still lacking. Herein we describe the development of a novel immunochemical approach to directly profile oxidized phosphatases. This immunochemical approach consists of an antibody designed to recognize the conserved sequence of the PTP active site (VHCDMDSAG) harboring the catalytic cysteine modified with dimedone (CDMD), a nucleophile that chemoselectively reacts with cysteine sulfenic acids to form a stable thioether adduct. Additionally, we provide biochemical and mass spectrometry workflows to be used in conjugation with this newly developed immunochemical approach to assist in the identification and quantification of basal and oxidized phosphatases.

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The Biological Buffer Bicarbonate/CO₂ Potentiates H₂O₂-Mediated Inactivation of Protein Tyrosine Phosphatases

Cited by 70 publications

References 33 publications

The molecular mechanisms and physiological consequences of oxidative stress: lessons from a model bacterium

The molecular mechanisms and physiological consequences of oxidative stress: lessons from a model bacterium

Hydrogen Peroxide in Biocatalysis. A Dangerous Liaison

An immunochemical approach to detect oxidized protein tyrosine phosphatases using a selective C-nucleophile tag

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