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Peroxynitrite formation in nitric oxide-exposed submitochondrial particles: detection, oxidative damage and catalytic removal by Mn-porphyrins.

Publication ,  Journal Article
Valez, V; Cassina, A; Batinic-Haberle, I; Kalyanaraman, B; Ferrer-Sueta, G; Radi, R
Published in: Arch Biochem Biophys
January 1, 2013

Peroxynitrite (ONOO(-)) formation in mitochondria may be favored due to the constant supply of superoxide radical (O(2)(∙-)) by the electron transport chain plus the facile diffusion of nitric oxide ((∙)NO) to this organelle. Herein, a model system of submitochondrial particles (SMP) in the presence of succinate plus the respiratory inhibitor antimycin A (to increase O(2)(∙-) rates) and the (∙)NO-donor NOC-7 was studied to directly establish and quantitate peroxynitrite by a multiplicity of methods including chemiluminescence, fluorescence and immunochemical analysis. While all the tested probes revealed peroxynitrite at near stoichiometric levels with respect to its precursor radicals, coumarin boronic acid (a probe that directly reacts with peroxynitrite) had the more straightforward oxidation profile from O(2)(∙-)-forming SMP as a function of the (∙)NO flux. Interestingly, immunospintrapping studies verified protein radical generation in SMP by peroxynitrite. Substrate-supplemented SMP also reduced Mn(III)porphyrins (MnP) to Mn(II)P under physiologically-relevant oxygen levels (3-30 μM); then, Mn(II)P were capable to reduce peroxynitrite and protect SMP from the inhibition of complex I-dependent oxygen consumption and protein radical formation and nitration of membranes. The data directly support the formation of peroxynitrite in mitochondria and demonstrate that MnP can undergo a catalytic redox cycle to neutralize peroxynitrite-dependent mitochondrial oxidative damage.

Duke Scholars

Published In

Arch Biochem Biophys

DOI

EISSN

1096-0384

Publication Date

January 1, 2013

Volume

529

Issue

1

Start / End Page

45 / 54

Location

United States

Related Subject Headings

  • Superoxides
  • Succinic Acid
  • Submitochondrial Particles
  • Spectrometry, Fluorescence
  • Rats
  • Peroxynitrous Acid
  • Oxygen
  • Oxidation-Reduction
  • Nitric Oxide Donors
  • Nitric Oxide
 

Citation

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Valez, V., Cassina, A., Batinic-Haberle, I., Kalyanaraman, B., Ferrer-Sueta, G., & Radi, R. (2013). Peroxynitrite formation in nitric oxide-exposed submitochondrial particles: detection, oxidative damage and catalytic removal by Mn-porphyrins. Arch Biochem Biophys, 529(1), 45–54. https://doi.org/10.1016/j.abb.2012.10.012
Valez, Valeria, Adriana Cassina, Ines Batinic-Haberle, Balaraman Kalyanaraman, Gerardo Ferrer-Sueta, and Rafael Radi. “Peroxynitrite formation in nitric oxide-exposed submitochondrial particles: detection, oxidative damage and catalytic removal by Mn-porphyrins.Arch Biochem Biophys 529, no. 1 (January 1, 2013): 45–54. https://doi.org/10.1016/j.abb.2012.10.012.
Valez V, Cassina A, Batinic-Haberle I, Kalyanaraman B, Ferrer-Sueta G, Radi R. Peroxynitrite formation in nitric oxide-exposed submitochondrial particles: detection, oxidative damage and catalytic removal by Mn-porphyrins. Arch Biochem Biophys. 2013 Jan 1;529(1):45–54.
Valez, Valeria, et al. “Peroxynitrite formation in nitric oxide-exposed submitochondrial particles: detection, oxidative damage and catalytic removal by Mn-porphyrins.Arch Biochem Biophys, vol. 529, no. 1, Jan. 2013, pp. 45–54. Pubmed, doi:10.1016/j.abb.2012.10.012.
Valez V, Cassina A, Batinic-Haberle I, Kalyanaraman B, Ferrer-Sueta G, Radi R. Peroxynitrite formation in nitric oxide-exposed submitochondrial particles: detection, oxidative damage and catalytic removal by Mn-porphyrins. Arch Biochem Biophys. 2013 Jan 1;529(1):45–54.
Journal cover image

Published In

Arch Biochem Biophys

DOI

EISSN

1096-0384

Publication Date

January 1, 2013

Volume

529

Issue

1

Start / End Page

45 / 54

Location

United States

Related Subject Headings

  • Superoxides
  • Succinic Acid
  • Submitochondrial Particles
  • Spectrometry, Fluorescence
  • Rats
  • Peroxynitrous Acid
  • Oxygen
  • Oxidation-Reduction
  • Nitric Oxide Donors
  • Nitric Oxide