Serial review: nitric oxide in mitochondria
Peroxynitrite reactions and formation in mitochondria

https://doi.org/10.1016/S0891-5849(02)01111-5Get rights and content

Abstract

Mitochondria constitute a primary locus for the intracellular formation and reactions of peroxynitrite, and these interactions are recognized to contribute to the biological and pathological effects of both nitric oxide (NO) and peroxynitrite. Extra- or intramitochondrially formed peroxynitrite can diffuse through mitochondrial compartments and undergo fast direct and free radical-dependent target molecule reactions. These processes result in oxidation, nitration, and nitrosation of critical components in the matrix, inner and outer membrane, and intermembrane space. Mitochondrial scavenging and repair systems for peroxynitrite-dependent oxidative modifications operate but they can be overwhelmed under enhanced cellular NO formation as well as under conditions that lead to augmented superoxide formation by the electron transport chain. Peroxynitrite can lead to alterations in mitochondrial energy and calcium homeostasis and promote the opening of the permeability transition pore. The effects of peroxynitrite in mitochondrial physiology can be largely rationalized based on the reactivities of peroxynitrite and peroxynitrite-derived carbonate, nitrogen dioxide, and hydroxyl radicals with critical protein amino acids and transition metal centers of key mitochondrial proteins. In this review we analyze (i) the existing evidence for the intramitochondrial formation and reactions of peroxynitrite, (ii) the key reactions and fate of peroxynitrite in mitochondria, and (iii) their impact in mitochondrial physiology and signaling of cell death.

Introduction

Mitochondria constitute a primary locus for the intracellular formation and reactions of peroxynitrite1, and these interactions are recognized to contribute to the biological and pathological effects of both nitric oxide (NO) and peroxynitrite. Peroxynitrite can diffuse from extramitochondrial compartments into the mitochondria or be formed intramitochondrially and undergo target molecule reactions with various constituents as well as react with carbon dioxide to yield secondary radicals. Altogether these reactions start one- and two-electron oxidation processes that result in the oxidation, nitration, and/or nitrosation of critical mitochondrial components that can lead to alterations in mitochondrial homeostasis and physiology. The concept of peroxynitrite interactions with mitochondria was originally proposed in 1994 [1]. The hypothesis was triggered by the realization that long-term inhibitory effects on cell respiration exerted by NO 2, 3 could not be solely explained by the rather limited direct reactions of NO with mitochondrial components, but was consistent with the participation of NO-derived secondary species with more robust chemical reactivity, such as peroxynitrite. In addition, as NO can freely diffuse through membranes and mitochondria is a key intracellular producer of superoxide radical anion (O2), the idea of the intramitochondrial formation of peroxynitrite from the rapid NO reaction with O2 was elaborated.

While it has been apparent that peroxynitrite formed in extramitochondrial sites can diffuse into and affect mitochondria, the intramitochondrial formation and reactions of peroxynitrite represents a more subtle issue, one that may contribute to explain the decreased biological half-life of NO under conditions that stimulate mitochondrial O2 formation, persistent effects of NO in mitochondrial functions, and mitochondrial signaling of NO-dependent cell death.

Section snippets

Peroxynitrite biochemistry and mitochondria

Peroxynitrite anion is formed from the diffusion-controlled reaction between the free radicals NO and O2 (k ∼ 1010 M−1s−1). Peroxynitrite anion (ONOO−) is in equilibrium with peroxynitrous acid (ONOOH, pKa = 6.8) and both species are strong oxidizing species in vivo by a variety of mechanisms that involve direct reactions with targets molecules (e.g., thiols, transition metal centers) or by secondary decomposition to free radicals, most notably carbonate (CO3) and nitrogen dioxide radicals

Peroxynitrite diffusion and mitochondria

The biological half-life of peroxynitrite in extracellular compartments has been estimated to be in the order of 10 ms, and both peroxynitrite anion and peroxynitrous acid can diffuse across biomembranes 6, 14, 15. Peroxynitrite formed outside mitochondria can diffuse to its interior, although the exact mechanisms remain to be elucidated; on the other hand, peroxynitrite formed intramitochondrially is expected to have a shorter biological half-life (t1/2 ∼ 3–5 ms) due to the large abundance of

In vitro and in vivo evidence supporting the mitochondrial formation/reactions of peroxynitrite

As peroxynitrite in mitochondria is a transient species, the evidence for its intramitochondrial formation mostly derives from (i) its reaction with exogenously added probes, (ii) the oxidative modifications of endogenous molecules (footprinting), and (iii) the SOD-inhibitable consumption and actions of NO. The existing evidence ranges from observations performed in submitochondrial particles (SMPs) to human tissues, as outlined in Table 1.

Peroxynitrite reactions with electron transport chain components

The effects of peroxynitrite are distinctly different to those of NO with mitochondrial electron transport chain components [48]. While NO mainly interacts and inhibits cytochrome c oxidase 29, 48, 49, 50, 51, 52, 53, 54, peroxynitrite reactions with complexes I, II, and V lead to their inactivation 1, 30, 48, 55, 56, 57, 58, 59, 60, by mechanisms that still need to be defined. This may not be an easy task, especially in the case of complexes I and II, as they are multicomponent complexes

Permeability transition pore (PTP) and the pyridine nucleotide-dependent calcium release pathway

Mitochondrial protein complexes that participate in critical mitochondrial/cellular processes such as the permeability transition pore (PTP) opening and the pyridine nucleotide-dependent calcium release pathway, contain inner membrane proteins with critical vicinal thiols that can be readily oxidized by peroxynitrite 77, 78. For the PTP, this protein is the adenine nucleotide translocase (ANT), known to be affected by oxidants including peroxynitrite 79, 80, 81. ANT can form a complex with the

Peroxynitrite interactions with matrix components

A number of matrix components are targets of peroxynitrite or peroxynitrite-derived radicals (Fig. 1). Metalloproteins such as aconitase and Mn-SOD present at 10–20 μM concentrations react with large rate constants with peroxynitrite, while glutathione present at ∼ 5 mM reacts at smaller rates and other compounds such as NADH only react with secondary radicals [6]. The relevant reaction of peroxynitrite with CO2 accounts for approximately half of the fate of peroxynitrite in the matrix and has

Overview of peroxynitrite reactions in mitochondria and their impact in cell homeostasis and death

We can then envision that peroxynitrite can either diffuse into mitochondria or be formed intramitochondrially and rapidly undergo target molecule reactions. All four mitochondrial compartments have interactions with peroxynitrite as VDAC, creatine kinase, ATPase, and aconitase, proteins present in the external membrane, intermembrane, inner membrane, and matrix respectively, are nitrated or oxidized during excess NO formation (Fig. 3).

Under physiological conditions, low fluxes of

Acknowledgements

This work was supported by grants from Fogarty-National Institutes of Health (USA) and the Howard Hughes Medical Institute (USA) to R.R. C.Q. is partially supported by a fellowship from PEDECIBA (Uruguay). R.R. is a Howard Hughes International Research Scholar.

References (115)

  • A. Boveris et al.

    Mitochondrial production of superoxide anions and its relationship to the antimycin insensitive respiration

    FEBS Lett

    (1975)
  • A. Okado-Matsumoto et al.

    Subcellular distribution of superoxide dismutases (SOD) in rat liverCu,Zn SOD in mitochondria

    J. Biol. Chem.

    (2001)
  • O. Stachowiak et al.

    Mitochondrial creatine kinase is a prime target of peroxynitrite-induced modification and inactivation

    J. Biol. Chem.

    (1998)
  • A. Boveris

    Determination of the production of superoxide radicals and hydrogen peroxide in mitochondria

    Methods Enzymol

    (1984)
  • T.E. Bates et al.

    Immunocytochemical evidence for a mitochondrially located nitric oxide synthase in brain and liver

    Biochem. Biophys. Res. Commun.

    (1995)
  • P. Ghafourifar et al.

    Nitric oxide synthase activity in mitochondria

    FEBS Lett

    (1997)
  • C. Giulivi et al.

    Production of nitric oxide by mitochondria

    J. Biol. Chem.

    (1998)
  • J.J. Poderoso et al.

    Nitric oxide inhibits electron transfer and increases superoxide radical production in rat heart mitochondria and submitochondrial particles

    Arch. Biochem. Biophys.

    (1996)
  • P. Ghafourifar et al.

    Mitochondrial nitric-oxide synthase stimulation causes cytochrome c release from isolated mitochondria. Evidence for intramitochondrial peroxynitrite formation

    J. Biol. Chem.

    (1999)
  • U. Bringold et al.

    Peroxynitrite formed by mitochondrial NO synthase promotes mitochondrial Ca2+ release

    Free Radic. Biol. Med.

    (2000)
  • L.A. Castro et al.

    Nitric oxide and peroxynitrite-dependent aconitase inactivation and iron-regulatory protein-1 activation in mammalian fibroblasts

    Arch. Biochem. Biophys.

    (1998)
  • W. Wu et al.

    Eosinophil peroxidase nitrates protein tyrosyl residues. Implications for oxidative damage by nitrating intermediates in eosinophilic inflammatory disorders

    J. Biol. Chem.

    (1999)
  • E. Linares et al.

    Role of peroxynitrite in macrophage microbicidal mechanisms in vivo revealed by protein nitration and hydroxylation

    Free Radic. Biol. Med.

    (2001)
  • F. Yamakura et al.

    Inactivation of human manganese-superoxide dismutase by peroxynitrite is caused by exclusive nitration of tyrosine 34 to 3-nitrotyrosine

    J. Biol. Chem.

    (1998)
  • C. Quijano et al.

    Reaction of peroxynitrite with Mn-superoxide dismutase. Role of the metal center in decomposition kinetics and nitration

    J. Biol. Chem.

    (2001)
  • L.A. MacMillan-Crow et al.

    Mitochondrial tyrosine nitration precedes chronic allograft nephropathy

    Free Radic. Biol. Med.

    (2001)
  • S. Raha et al.

    Mitochondria, oxygen free radicals, disease and ageing

    Trends Biochem. Sci.

    (2000)
  • A. Cassina et al.

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

    Arch. Biochem. Biophys.

    (1996)
  • M.W. Cleeter et al.

    Reversible inhibition of cytochrome c oxidase, the terminal enzyme of the mitochondrial respiratory chain, by nitric oxide. Implications for neurodegenerative diseases

    FEBS Lett

    (1994)
  • G.C. Brown et al.

    Nanomolar concentrations of nitric oxide reversibly inhibit synaptosomal respiration by competing with oxygen at cytochrome oxidase

    FEBS Lett

    (1994)
  • Y. Takehara et al.

    Oxygen-dependent regulation of mitochondrial energy metabolism by nitric oxide

    Arch. Biochem. Biophys.

    (1995)
  • G.C. Brown

    Regulation of mitochondrial respiration by nitric oxide inhibition of cytochrome c oxidase

    Biochim. Biophys. Acta

    (2001)
  • C. Szabo et al.

    Endogenous peroxynitrite is involved in the inhibition of mitochondrial respiration in immuno-stimulated J774.2 macrophages

    Biochem. Biophys. Res. Commun.

    (1995)
  • V. Borutaite et al.

    Reversal of nitric oxide-, peroxynitrite- and S-nitrosothiol-induced inhibition of mitochondrial respiration or complex I activity by light and thiols

    Biochim. Biophys. Acta

    (2000)
  • L.L. Pearce et al.

    Identification of respiratory complexes I and III as mitochondrial sites of damage following exposure to ionizing radiation and nitric oxide

    Nitric Oxide

    (2001)
  • H. Rubbo et al.

    Peroxynitrite inactivates thiol-containing enzymes of Trypanosoma cruzi energetic metabolism and inhibits cell respiration

    Arch. Biochem. Biophys.

    (1994)
  • Y. Kagawa

    Proton motive ATP synthesis

  • Y. Zhang et al.

    The oxidative inactivation of mitochondrial electron transport chain components and ATPase

    J. Biol. Chem.

    (1990)
  • L.L. Pearce et al.

    The peroxynitrite reductase activity of cytochrome c oxidase involves a two-electron redox reaction at the heme a(3)-Cu(B) site

    J. Biol. Chem.

    (1999)
  • M.A. Sharpe et al.

    Interaction of peroxynitrite with mitochondrial cytochrome oxidase. Catalytic production of nitric oxide and irreversible inhibition of enzyme activity

    J. Biol. Chem.

    (1998)
  • R. Radi et al.

    Cytochrome c-catalyzed membrane lipid peroxidation by hydrogen peroxide

    Arch. Biochem. Biophys.

    (1991)
  • L. Thomson et al.

    Kinetics of cytochrome c2+ oxidation by peroxynitriteimplications for superoxide measurements in nitric oxide-producing biological systems

    Arch. Biochem. Biophys.

    (1995)
  • A.M. Cassina et al.

    Cytochrome c nitration by peroxynitrite

    J. Biol. Chem.

    (2000)
  • V. Borutaite et al.

    Release of cytochrome c from heart mitochondria is induced by high Ca2+ and peroxynitrite and is responsible for Ca(2+)-induced inhibition of substrate oxidation

    Biochim. Biophys. Acta

    (1999)
  • J.F. Turrens et al.

    Ubisemiquinone is the electron donor for superoxide formation by complex III of heart mitochondria

    Arch. Biochem. Biophys.

    (1985)
  • L. Zhang et al.

    Generation of superoxide anion by succinate-cytochrome c reductase from bovine heart mitochondria

    J. Biol. Chem.

    (1998)
  • A.P. Halestrap et al.

    Oxidative stress, thiol reagents, and membrane potential modulate the mitochondrial permeability transition by affecting nucleotide binding to the adenine nucleotide translocase

    J. Biol. Chem.

    (1997)
  • J.L. Scarlett et al.

    Alterations to glutathione and nicotinamide nucleotides during the mitochondrial permeability transition induced by peroxynitrite

    Biochem. Pharmacol.

    (1996)
  • D. Brdiczka et al.

    In vitro complex formation between the octamer of mitochondrial creatine kinase and porin

    J. Biol. Chem.

    (1994)
  • G. Beutner et al.

    Complexes between kinases, mitochondrial porin and adenylate translocator in rat brain resemble the permeability transition pore

    FEBS Lett

    (1996)
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    Guest Editors: Christoph Richter and Matthias Schweizer

    This article is part of a series of reviews on “Nitric Oxide in Mitochondria.” The full list of papers may be found on the homepage of the journal.

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