Unraveling peroxynitrite formation in biological systems

doi:10.1016/S0891-5849(00)00373-7

Free Radical Biology and Medicine

Volume 30, Issue 5, 1 March 2001, Pages 463-488

https://doi.org/10.1016/S0891-5849(00)00373-7 Get rights and content

Abstract

Peroxynitrite promotes oxidative damage and is implicated in the pathophysiology of various diseases that involve accelerated rates of nitric oxide and superoxide formation. The unambiguous detection of peroxynitrite in biological systems is, however, difficult due to the combination of a short biological half-life, limited diffusion, multiple target molecule reactions, and participation of alternative oxidation/nitration pathways. In this review, we provide the conceptual framework and a comprehensive analysis of the current experimental strategies that can serve to unequivocally define the existence and quantitation of peroxynitrite in biological systems of different levels of organization and complexity.

Introduction

Peroxynitrite,¹ the product of the combination reaction between nitric oxide (^·NO) and superoxide (O₂^•−), is a reactive and short-lived species that promotes oxidative molecular and tissue damage [1], [2], [3], [4], [5], [6], [7]. In addition to the generation of a pro-oxidant species, the formation of peroxynitrite results in decreased bioavailability of radical dot NO, therefore diminishing both its salutary physiological functions [8], [9], [10] and its strong antioxidant actions over free radical and metal-mediated processes [11], [12], [13]. Peroxynitrite formation and reactions are proposed to contribute to the pathogenesis of a series of diseases including acute and chronic inflammatory processes, sepsis, ischemia-reperfusion, and neurodegenerative disorders, among others [14], [15], [16], [17], [18], [19], [20], [21], [22], [23], [24], [25], [26], [27].

The detection of peroxynitrite in biological systems has been a challenge over the past decade because of the (i) elusive nature of peroxynitrite which precludes its direct isolation and detection, (ii) necessity to find detector molecules that can efficiently outcompete the multiple reactions that peroxynitrite can undergo, (iii) nonexistence of footprints totally specific of peroxynitrite reactions, and (iv) the difficulty to discriminate between the biological effects of peroxynitrite versus that of its precursors, radical dot NO and O₂^•−, and other NO-derived oxidants.

In spite of the fact that the biological formation and reactions of peroxynitrite are kinetically and thermodynamically favored, the importance of the peroxynitrite pathway in biology has been occasionally questioned [28], [29], [30], in part due to the difficulties related to its detection. Even though there is solid evidence supporting the formation of peroxynitrite in vivo and its contribution to biomolecular damage and cell and tissue pathology, unambiguous detection and quantitation is not trivial and requires a subtle knowledge of its biological chemistry and a multifaceted approach. In this work, we will (i) provide biochemical and physico-chemical foundation needed to search for peroxynitrite, (ii) analyze current methodologies used in the detection of peroxynitrite and (iii) establish criteria that must be fulfilled to unravel the formation of peroxynitrite in biological systems of different levels of organization and complexity.

Section snippets

Formation reactions

The biological formation of peroxynitrite anion (ONOO⁻) is mainly due to the fast reaction between radical dot NO and O₂^•− (Eqn. 1). This radical-radical combination reaction undergoes with a second order rate constant that has been independently determined as 4.3, 6.7, and 19 × 10⁹ M⁻¹ s⁻¹ [31], [32], [33], and therefore one can safely assume a value of ∼10¹⁰ M⁻¹s⁻¹, which indicates a diffusion-controlled reaction. $^{•} NO+O_{2}^{•−} →ONOO^{−} v=k[^{•} NO][O_{2}^{•−}]$

Since both precursor radical species, radical dot NO and O₂^•−, are

General considerations

The detection of peroxynitrite relies on either (i) modification of exogenously added probes, or (ii) footprinting reactions on endogenous molecules. However, these are not straightforward procedures; at present there are no totally specific modifications of either probe or biomolecules that can directly and unambiguously assure the formation of peroxynitrite. Probe modification and/or footprinting reactions require additional criteria to constitute sufficient evidence for affirming

Methodologies for peroxynitrite detection

In this section we will describe current methodologies for peroxynitrite detection. The analysis will concentrate on those techniques that have been more widely used and validated. We will provide the biochemical background for each methodology, its potency and limitations. The field is in progress and awaits further application and development.

Pharmacology to unravel peroxynitrite

Due to the lack of techniques and chemical modifications completely specific for peroxynitrite, additional experimental evidence is obtained by the use of: (i) drugs that decrease radical dot NO and O₂^•− levels, (ii) treatments that modify the concentration of biomolecules that critically influence peroxynitrite reactivity, (iii) peroxynitrite decomposition catalysts and scavengers, and (iv) compounds that interfere with alternative oxidation/nitration pathways. Finally, in some models it may be of

Conclusions

•
Nitric oxide-superoxide interactions readily occur in vivo leading to the formation of peroxynitrite.
•
Peroxynitrite is short-lived, therefore its detection relies on modification of exogenous detector (probe oxidation) or endogenous target (footprinting) molecules.
•
Peroxynitrite-induced modifications, most notably oxidations and nitrations, can be followed in biological systems of different levels of complexity including biochemical, cellular, tissue, and organ levels.
•
Various methods are

Acknowledgements

This work was supported by grants from ICGEB (Trieste), SAREC (Sweden), Fogarty-NIH (USA), CONICYT and CSIC (Uruguay) to R.R. We thank Drs. Ohara Augusto and Jay W. Heinecke for helpful comments during the preparation of the manuscript and Dr. Gabriela Gualco for her advice on immunohistopathology.

References (200)

R. Radi et al.
Peroxynitrite oxidation of sulfhydryls. The cytotoxic potential of superoxide and nitric oxide
J. Biol. Chem.
(1991)
R. Radi et al.
Peroxynitrite-induced membrane lipid peroxidationthe cytotoxic potential of superoxide and nitric oxide
Arch. Biochem. Biophys.
(1991)
H. Ischiropoulos et al.
Peroxynitrite-mediated tyrosine nitration catalyzed by superoxide dismutase
Arch. Biochem. Biophys.
(1992)
H. Rubbo et al.
Nitric oxide regulation of superoxide and peroxynitrite-dependent lipid peroxidation. Formation of novel nitrogen-containing oxidized lipid derivatives
J. Biol. Chem.
(1994)
H. Kaur et al.
Evidence for nitric oxide-mediated oxidative damage in chronic inflammation. Nitrotyrosine in serum and synovial fluid from rheumatoid patients
FEBS Lett.
(1994)
N. Fukuyama et al.
Clinical evidence of peroxynitrite formation in chronic renal failure patients with septic shock
Free Radic. Biol. Med.
(1997)
D.A. Wink et al.
Chemical biology of nitric oxideinsights into regulatory, cytotoxic and cytoprotective mechanisms of nitric oxide
Free Radic. Biol. Med.
(1998)
S. Goldstein et al.
The reaction of NO• with O2•− and HO2•a pulse radiolysis study
Free Radic. Biol. Med.
(1995)
A. Denicola et al.
Nitric oxide diffusion in membranes determined by fluorescence quenching
Arch. Biochem. Biophys.
(1996)
M.N. Hughes
Relationships between nitric oxide, nitroxyl ion, nitrosonium cation and peroxynitrite
Biochim. Biophys. Acta
(1999)

A. Denicola et al.

Peroxynitrite reaction with carbon dioxide/bicarbonatekinetics and influence on peroxynitrite-mediated oxidations

Arch. Biochem. Biophys.

(1996)

M.G. Bonini et al.

Direct epr detection of the carbonate radical anion produced from peroxynitrite and carbon dioxide

J. Biol. Chem.

(1999)

O. Augusto et al.

Spin-trapping studies of peroxynitrite decomposition and of 3-morpholinosydnonimine N-ethylcarbamide autooxidationdirect evidence for metal-independent formation of free radical intermediates

Arch. Biochem. Biophys.

(1994)

H. Ischiropoulos

Biological tyrosine nitrationa pathophysiological function of nitric oxide and reactive oxygen species

Arch. Biochem. Biophys.

(1998)

B. Alvarez et al.

Kinetics of peroxynitrite reaction with amino acids and human serum albumin

J. Biol. Chem.

(1999)

S. Goldstein et al.

Tyrosine nitration by simultaneous generation of (•)NO and O-(2) under physiological conditions. How the radicals do the job

J. Biol. Chem.

(2000)

A. van der Vliet et al.

Tyrosine modification by reactive nitrogen speciesa closer look

Arch. Biochem. Biophys.

(1995)

N. Romero et al.

Diffusion of peroxynitrite in the presence of carbon dioxide

Arch. Biochem. Biophys.

(1999)

R. Radi et al.

Inhibition of mitochondrial electron transport by peroxynitrite

Arch. Biochem. Biophys.

(1994)

A. Cassina et al.

Differential inhibitory action of nitric oxide and peroxynitrite on mitochondrial electron transport

Arch. Biochem. Biophys.

(1996)

G.L. Squadrito et al.

Oxidative chemistry of nitric oxidethe roles of superoxide, peroxynitrite and carbon dioxide

Free Radic. Biol. Med.

(1998)

M.R. Gunther et al.

Nitric oxide trapping of the tyrosyl radical of prostaglandin H synthase-2 leads to tyrosine iminoxyl radical and nitrotyrosine formation

J. Biol. Chem.

(1997)

J.A. Royall et al.

Evaluation of 2′,7′-dichlorofluorescin and dihydrorhodamine 123 as fluorescent probes for intracellular H2O2 in cultured endothelial cells

Arch. Biochem. Biophys.

(1993)

N.W. Kooy et al.

Peroxynitrite-mediated oxidation of dihydrorhodamine 123

Free Radic. Biol. Med.

(1994)

J.P. Crow

Dichlorodihydrofluorescein and dihydrorhodamine 123 are sensitive indicators of peroxynitrite in vitroimplications for intracellular measurement of reactive nitrogen and oxygen species

Nitric Oxide

(1997)

H. Ischiropoulos et al.

Detection of reactive nitrogen species using 2,7-dichlorodihydrofluorescein and dihydrorhodamine 123

Methods Enzymol.

(1999)

A.M. Miles et al.

Modulation of superoxide-dependent oxidation and hydroxylation reactions by nitric oxide

J. Biol. Chem.

(1996)

C. Rota et al.

Evidence for free radical formation during the oxidation of 2′-7′-dichlorofluorescin to the fluorescent dye 2′-7′-dichlorofluorescein by horseradish peroxidasepossible implications for oxidative stress measurements

Free Radic. Biol. Med.

(1999)

C. Rota et al.

Phenoxyl free radical formation during the oxidation of the fluorescent dye 2′,7′-dichlorofluorescein by horseradish peroxidase. Possible consequences for oxidative stress measurements

J. Biol. Chem.

(1999)

H. Possel et al.

2,7-Dihydrodichlorofluorescein diacetate as a fluorescent marker for peroxynitrite formation

FEBS Lett.

(1997)

C. Gagnon et al.

Peroxynitrite production by human neutrophils, monocytes and lymphocytes challenged with lipopolysaccharide

FEBS Lett.

(1998)

M.P. Lopez-Garcia et al.

Peroxynitrite generated from constitutive nitric oxide synthase mediates the early biochemical injury in short-term cultured hepatocytes

FEBS Lett.

(2000)

N.W. Kooy et al.

Agonist-induced peroxynitrite production from endothelial cells

Arch. Biochem. Biophys.

(1994)

S.D. Catz et al.

Nitric oxide synthase inhibitors decrease human polymorphonuclear leukocyte luminol-dependent chemiluminescence

Free Radic. Biol. Med.

(1995)

L. Castro et al.

Modulatory role of nitric oxide on superoxide-dependent luminol chemiluminescence

Arch. Biochem. Biophys.

(1996)

K. Faulkner et al.

Luminol and lucigenin as detectors for O2^•−

Free Radic. Biol. Med.

(1993)

H. Ischiropoulos et al.

Peroxynitrite formation from macrophage-derived nitric oxide

Arch. Biochem. Biophys.

(1992)

S. Pfeiffer et al.

Dityrosine formation outcompetes tyrosine nitration at low steady-state concentrations of peroxynitrite. Implications for tyrosine modification by nitric oxide/superoxide in vivo

J. Biol. Chem.

(2000)

J.P. Crow et al.

Detection and quantitation of nitrotyrosine residues in proteinsin vivo marker of peroxynitrite

Methods Enzymol.

(1996)

J.P. Crow

Measurement and significance of free and protein-bound 3-nitrotyrosine, 3-chlorotyrosine, and free 3-nitro-4-hydroxyphenylacetic acid in biologic samplesa high-performance liquid chromatography method using electrochemical detection

Methods Enzymol.

(1999)

C. Herce-Pagliai et al.

Analytical methods for 3-nitrotyrosine as a marker of exposure to reactive nitrogen speciesa review

Nitric Oxide

(1998)

M.K. Shigenaga

Quantitation of protein-bound 3-nitrotyrosine by high-performance liquid chromatography with electrochemical detection

Methods Enzymol.

(1999)

A. van der Vliet et al.

Nitrotyrosine as biomarker for reactive nitrogen species

Methods Enzymol.

(1996)

S. Pennathur et al.

Mass spectrometric quantification of 3-nitrotyrosine, ortho-tyrosine, and o,o′-dityrosine in brain tissue of 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine-treated mice, a model of oxidative stress in Parkinson’s disease

J. Biol. Chem.

(1999)

J.R. Crowley et al.

Isotope dilution mass spectrometric quantification of 3-nitrotyrosine in proteins and tissues is facilitated by reduction to 3-aminotyrosine

Anal. Biochem.

(1998)

H. Jiang et al.

Detection of 3-nitrotyrosine in human platelets exposed to peroxynitrite by a new gas chromatography/mass spectrometry assay

Nitric Oxide

(1998)

E. Schwedhelm et al.

Gas chromatographic-tandem mass spectrometric quantification of free 3-nitrotyrosine in human plasma at the basal state

Anal. Biochem.

(1999)

J.S. Beckman et al.

Apparent hydroxyl radical production by peroxynitriteimplications for endothelial injury from nitric oxide and superoxide

Proc. Natl. Acad. Sci. USA

(1990)

W.A. Pryor et al.

The chemistry of peroxynitritea product from the reaction of nitric oxide with superoxide

Am. J. Physiol.

(1995)

J.S. Beckman et al.

Nitric oxide, superoxide, and peroxynitritethe good, the bad, and ugly

Am. J. Physiol.

(1996)

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²: Rafael Radi, M.D., Ph.D., obtained his doctoral degree at the Universidad de la República, Montevideo, Uruguay in 1989 and performed posdoctoral studies at the University of Alabama at Birmingham (1989–1991). He returned to Uruguay in 1992 to a faculty position at the Departamento de Bioquı́mica, Facultad de Medicina, Universidad de la República, where he initiated a research group that investigates the biochemistry and cell biology of nitric oxide and peroxynitrite. He is at present a Professor of Biochemistry, an International Research Scholar of the Howard Hughes Medical Institute, and the current Secretary General of the Oxygen Society.

³: Gonzalo Peluffo, M.D., and Marı́a Noel Alvarez, M.S., obtained their degrees at the Universidad de la República in 1998 and are currently performing Ph.D. studies on aspects referred to the biological formation, detection, and diffusion of oxygen radicals, nitric oxide, and peroxynitrite in biochemical and cell systems (MNA) or humans (GP). They are both Assistant Professors of Biochemistry at the Facultad de Medicina, Universidad de la República.

⁴: Mercedes Naviliat, M.D., obtained her degree at the Universidad de la República in 1985 and has just completed a Ph.D. thesis on the role of nitric oxide and peroxynitrite in inflammatory disease. She is an Assistant Professor of Rheumatology at the Facultad de Medicina, Universidad de la República.

⁵: Alfonso Cayota, M.D., Ph.D., obtained his M.D. degree at Universidad de la República in 1986 and performed Ph.D. studies at the Université Paris VI-Pasteur Institute, Paris, France from 1990–1995. He is currently an Associate Professor of Medicine at the Facultad de Medicina, Universidad de la República, where he investigates the role of reactive oxygen and nitrogen species during normal and pathological immune responses.

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Forum: oxidative stress statusUnraveling peroxynitrite formation in biological systems

Abstract

Introduction

Section snippets

Formation reactions

General considerations

Methodologies for peroxynitrite detection

Pharmacology to unravel peroxynitrite

Conclusions

Acknowledgements

J. Biol. Chem.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

J. Biol. Chem.

FEBS Lett.

Free Radic. Biol. Med.

Free Radic. Biol. Med.

Free Radic. Biol. Med.

Arch. Biochem. Biophys.

Biochim. Biophys. Acta

Arch. Biochem. Biophys.

J. Biol. Chem.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

J. Biol. Chem.

J. Biol. Chem.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

Free Radic. Biol. Med.

J. Biol. Chem.

Arch. Biochem. Biophys.

Free Radic. Biol. Med.

Nitric Oxide

Methods Enzymol.

J. Biol. Chem.

Free Radic. Biol. Med.

J. Biol. Chem.

FEBS Lett.

FEBS Lett.

FEBS Lett.

Arch. Biochem. Biophys.

Free Radic. Biol. Med.

Arch. Biochem. Biophys.

Free Radic. Biol. Med.

Arch. Biochem. Biophys.

J. Biol. Chem.

Methods Enzymol.

Methods Enzymol.

Nitric Oxide

Methods Enzymol.

Methods Enzymol.

J. Biol. Chem.

Anal. Biochem.

Nitric Oxide

Anal. Biochem.

Apparent hydroxyl radical production by peroxynitriteimplications for endothelial injury from nitric oxide and superoxide

Proc. Natl. Acad. Sci. USA

The chemistry of peroxynitritea product from the reaction of nitric oxide with superoxide

Am. J. Physiol.

Nitric oxide, superoxide, and peroxynitritethe good, the bad, and ugly

Am. J. Physiol.

Forum: oxidative stress status
Unraveling peroxynitrite formation in biological systems