Modulation of catalase peroxidatic and catalatic activity by nitric oxide

doi:10.1016/S0891-5849(00)00512-8

Free Radical Biology and Medicine

Volume 30, Issue 7, 1 April 2001, Pages 709-714

https://doi.org/10.1016/S0891-5849(00)00512-8 Get rights and content

Abstract

Previously, we found that catalase enhanced the protection afforded by superoxide dismutase to Escherichia coli against the simultaneous generation of superoxide and nitric oxide (Brunelli et al., Arch. Biochem. Biophys. 316:327–334, 1995). Hydrogen peroxide itself was not toxic in this system in the presence or absence of superoxide dismutase. We therefore investigated whether catalase might consume nitric oxide in addition to hydrogen peroxide. Catalase rapidly formed a reversible complex stoichiometrically with nitric oxide with the Soret band shifting from 406 to 426 nm and two new peaks appeared at 540 and at 575 nm, consistent with the formation of a ferrous-nitrosyl complex. Catalase consumed more nitric oxide upon the addition of hydrogen peroxide. Conversely, micromolar concentrations of nitric oxide slowed the catalase-mediated decomposition of hydrogen peroxide. Catalase pretreated with nitric oxide and hydrogen peroxide regained full activity after dialysis. Our results suggest that catalase can slowly consume nitric oxide while nitric oxide modestly inhibits catalase-dependent scavenging of hydrogen peroxide. The protective effects of catalase in combination with superoxide dismutase may result from two actions; reducing peroxynitrite formation by scavenging nitric oxide and by scavenging hydrogen peroxide before it reacts with superoxide dismutase to form additional superoxide.

Introduction

Catalase is a four subunits 240 kDa ferric hemoprotein that mediates the two step decomposition of hydrogen peroxide (H₂O₂) to water [1]. This is commonly referred to as the catalatic activity of catalase and occurs according to the following pathway: $catalase-Fe^{3+} +H_{2} O_{2} +2H^{+} ⇒compound I (Fe^{4+} =O)+H_{2} O$ $compound I (Fe^{4+} =O)+H_{2} O_{2} ⇒catalase-Fe^{3+} +H_{2} O+O_{2}$

Catalase also demonstrates a peroxidatic activity toward small substrates, such as methanol, ethanol, formate, and azide as first reported by Keilin and Hartree [2], [3]. The essential feature of the peroxidatic activity of catalase is the oxidation of these alternative substrates in competition with hydrogen peroxide for compound I (reaction 2). Therefore, the peroxidatic activity is most evident at relatively low concentrations of hydrogen peroxide [3], [4]. Catalase prefers small substrates because the heme groups are deeply buried and are accessible only by a narrow channel lined with hydrophobic residues [5].

Nitric oxide ( radical dot NO) is a small hydrophobic gas that has physiologic functions as diverse as smooth muscle relaxation, platelet inhibition, neurotransmission, and stimulation of hormone release [6], [7], [8]. The information carried by nitric oxide is translated into a cellular signal by reversibly binding to the ferrous heme iron of guanylate cyclase that stimulates cGMP synthesis [7]. However, nitric oxide can also react with ferric hemoproteins such as ferrimyoglobin and ferric horseradish peroxidase [9].

The restrictive active site of catalase is large enough to allow ready access to nitric oxide. Catalase is known to form a transient complex with nitric oxide when it oxidizes azide in the presence of hydrogen peroxide [10]. We became interested in the interactions of nitric oxide with catalase when we observed that catalase significantly decreased the bactericidal activity of 3-morpholinosydnonimine-N-ethylcarbamide (SIN-1), a compound that produces peroxynitrite (ONOO⁻) through the release of nitric oxide and superoxide (O₂^•−) [11]. Therefore, we investigated the effects of nitric oxide on the catalase-mediated scavenging of hydrogen peroxide and whether nitric oxide might be consumed by the peroxidatic activity of catalase.

Section snippets

Materials and methods

A 1.9 mM nitric oxide solution was prepared by bubbling 100 ml of water in a 300 ml gas sampling cylinder (Fisher Scientific, Fair Lawn, NJ, USA) with argon for 15 min and then with nitric oxide for 5 min. Catalase was purchased from Worthington Biochemical Corp., Freehold, NJ, USA. Hydrogen peroxide (30%) was obtained from Fisher Scientific. Catalase was dialyzed against 50 mM potassium phosphate, pH 7.0, and filter sterilized. Spectral analysis was carried out with either a UV-260 Shimadzu or

Results

The Soret band of native catalase peak shifted from 406 to 426 nm with an isosbestic point at 415 nm when 20 μM nitric oxide was added to the catalase solution under either aerobic or anaerobic conditions (Fig. 1). Two new peaks appeared at 540 nm and at 575 nm. Such spectral changes are in agreement with those reported by Nicholls for the nitric oxide-catalase complex and are typical of a ferrous-nitrosyl complex [10]. Nitric oxide was then removed from the catalase solution by bubbling

Discussion

Catalase readily forms a complex with nitric oxide, and Brown [13] showed that submicromolar concentrations of nitric oxide inhibited oxygen evolution by catalase. In the present study, we showed that nitric oxide in addition substantially slowed the overall consumption of hydrogen peroxide. Oxygen evolution only assesses the second step in the catalase cycle (reaction 2) and is competitively inhibited by alternative peroxidative substrates like ethanol. However, many peroxidative substrates

Acknowledgements

We thank Dr. Irwin Fridovich for providing laboratory space, equipment and many helpful discussions. Dr. Brunelli thanks Prof. Alberto Bertolini and Prof. Alberto Ponzone for their support.

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Properties of catalase. Catalysis of coupled oxidation of alcohols

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Protein nitration induced by Hemin/NO: A complementary mechanism through the catalytic functions of hemin and NO-scavenging
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Hemin and heme-peroxidases have been considered essential catalysts for the nitrite/hydrogen peroxide (H₂O₂)-mediated protein nitration in vitro, understood as one of the main pathways for protein modification in biological systems. However, the role of nitric oxide (^●NO) in the heme/hemin-induced protein nitration has not been studied in-depth. This is despite its reductive nitrosylating effects following binding to hemin and the possible involvement of the reactive nitrogen species in the nitration of various functional proteins. Here, the ^●NO-binding affinity of hemin has been studied along with the influence of ^●NO on the internalization of hemin into MDA-MB-231 cells and the accompanying changes in the profile of intracellular nitrated proteins. Moreover, to further understand the mechanism involved, bovine serum albumin (BSA) nitration was studied after treatment with hemin and ^●NO, with an investigation of the effects of pH of the reaction medium, generation of H₂O₂, and the oxidation of the tyrosine residues as the primary sites for the nitration. We demonstrated that hemin nitrosylation enhanced its cellular uptake and induced the one-electron oxidation and nitration of different intracellular proteins along with its ^●NO-scavenging efficiency. Moreover, the hemin/NO-mediated BSA nitration was proved to be dependent on the concentration of ^●NO and the pH of the reaction medium, with a vital role being played by the scavenging effects of protein for the free hemin molecules. Collectively, our results reaffirm the involvement of hemin and ^●NO in the nitration mechanism, where the nitrosylation products can induce protein nitration while promoting the effects of the components of the nitrite/H₂O₂-mediated pathway.
Nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and Fe(II) Fenton reactants in E. coli
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Dioxygen activating heme enzymes have long predicted to be powerhouses for nitrogen oxide interconversion, especially for nitric oxide (NO) oxidation which has far-reaching biological and/or environmental impacts. Lending credence, reactivity of NO with high-valent heme‑oxygen intermediates of globin proteins has recently been implicated in the regulation of a variety of pivotal physiological events such as modulating catalytic activities of various heme enzymes, enhancing antioxidant activity to inhibit oxidative damage, controlling inflammatory and infectious properties within the local heme environments, and NO scavenging. To reveal insights into such crucial biological processes, we have investigated low temperature NO reactivities of two classes of synthetic high-valent heme intermediates, Compound-II and Compound-I. In that, Compound-II rapidly reacts with NO yielding the six-coordinate (NO bound) heme ferric nitrite complex, which upon warming to room temperature converts into the five-coordinate heme ferric nitrite species. These ferric nitrite complexes mediate efficient substrate oxidation reactions liberating NO; i.e., shuttling NO₂⁻ back to NO. In contrast, Compound-I and NO proceed through an oxygen-atom transfer process generating the strong nitrating agent NO₂, along with the corresponding ferric nitrosyl species that converts to the naked heme ferric parent complex upon warmup. All reaction components have been fully characterized by UV–vis, ²H NMR and EPR spectroscopic methods, mass spectrometry, elemental analyses, and semi-quantitative determination of NO₂⁻ anions. The clean, efficient, potentially catalytic NO_x interconversions driven by high-valent heme species presented herein illustrate the strong prospects of a heme enzyme/O₂/NO_x dependent unexplored territory that is central to human physiology, pathology, and therapeutics.
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Free Radical Biology and Medicine

Abstract

Introduction

Section snippets

Materials and methods

Results

Discussion

Acknowledgements

References (22)

The comparative toxicity of nitric oxide and peroxynitrite to Escherichia coli

Arch. Biochem. Biophys.

Direct measurement of nitric oxide in solutions with an ozone based chemiluminescent detector

Methods

Superoxide radical inhibits catalase

J. Biol. Chem.

Increased cytotoxicity of 3-morpholinosydnonimine to HepG2 cells in the presence of superoxide dismutase

J. Biol. Chem.

Bactericidal agents generated by the peroxidase-catalyzed oxidation of para-hydroquinones

J. Biol. Chem.

On the role of bicarbonate in peroxidations catalyzed by Cu,Zn superoxide dismutase

Free Radic. Biol. Med.

Bicarbonate enhances the hydroxylation, nitration, and peroxidation reactions catalyzed by copper, zinc superoxide dismutase

J. Biol. Chem.

Superoxide dismutase and hydrogen peroxide cause rapid nitric oxide breakdown, peroxynitrite production and subsequent cell death

Biochim. Biophys. Acta

An intermediate compound in the catalase-hydrogen peroxide reaction

Acta Chem. Scand.

Coupled oxidation of alcohol

Proc. R. Soc. Lond. B Biol. Sci.

Properties of catalase. Catalysis of coupled oxidation of alcohols

Biochem. J.

Cited by (125)

Compromised conformation and kinetics of catalase in the presence of propylthiouracil: A biophysical study and alleviation by curcumin

Protein nitration induced by Hemin/NO: A complementary mechanism through the catalytic functions of hemin and NO-scavenging

Nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and Fe(II) Fenton reactants in E. coli

Bio-inspired nitrogen oxide (NO<inf>x</inf>) interconversion reactivities of synthetic heme Compound-I and Compound-II intermediates

Phytotoxicity of the chiral herbicide dichlorprop: Cross-talk between nitric oxide, reactive oxygen species and phytohormones

Browsing the oldest antioxidant enzyme: catalase and its multiple regulation in cancer

Original contributionModulation of catalase peroxidatic and catalatic activity by nitric oxide

Abstract

Introduction

Section snippets

Materials and methods

Results

Discussion

Acknowledgements

Arch. Biochem. Biophys.

Methods

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Free Radic. Biol. Med.

J. Biol. Chem.

Biochim. Biophys. Acta

An intermediate compound in the catalase-hydrogen peroxide reaction

Acta Chem. Scand.

Coupled oxidation of alcohol

Proc. R. Soc. Lond. B Biol. Sci.

Properties of catalase. Catalysis of coupled oxidation of alcohols

Biochem. J.

Original contribution
Modulation of catalase peroxidatic and catalatic activity by nitric oxide