Cited by 293Electron Spin Resonance StudiesRedox biology and oxidative stressFree Radicals and Antioxidants

Direct EPR Detection of the Carbonate Radical Anion Produced from Peroxynitrite and Carbon Dioxide

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Bonini, Marcelo G., Radi, Rafael, Ferrer-Sueta, Gerardo, Ferreira, Ana M. Da C., Augusto, Ohara·1999·Journal of Biological Chemistry

Abstract

The biological effects of peroxynitrite have been recently considered to be largely dependent on its reaction with carbon dioxide, which is present in high concentrations in intra- and extracellular compartments. Peroxynitrite anion (ONOO-) reacts rapidly with carbon dioxide, forming an adduct, nitrosoperoxocarboxylate (ONOOCO2-), whose decomposition has been proposed to produce reactive intermediates such as the carbonate radical (CO-3). Here, by the use of rapid mixing continuous flow electron paramagnetic resonance (EPR), we directly detected the carbonate radical in flow mixtures of peroxynitrite with bicarbonate-carbon dioxide over the pH range of 6-9. The radical was unambiguously identified by its EPR parameters (g = 2.0113; line width = 5.5 G) and by experiments with bicarbonate labeled with 13C. In this case, the singlet EPR signal obtained with 12C bicarbonate splits into the expected doublet because of 13C (a(13C)= 11.7 G). The singlet spectrum of the unlabeled radical was invariant between pH 6 and 9, confirming that in this pH range the detected radical is the carbonate radical anion (CO-3). Importantly, in addition to contributing to the understanding of nitrosoperoxocarboxylate decomposition pathways, this is the first report unambiguously demonstrating the formation of the carbonate radical anion at physiological pHs by direct EPR spectroscopy.

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Crossref · 10.1074/jbc.274.16.10802

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  • The carbonate radical can be formed in vivo by the reaction of the hydroxyl radical with carbonate or bicarbonate or via the reaction of peroxynitrite with carbon dioxide.

    “The carbonate radical can be formed in vivo by the reaction of the hydroxyl radical with carbonate or bicarbonate (138) or via the reaction of peroxynitrite with carbon dioxide (Eq. 30) (112, 608, 640).”

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  1. [3]3. Beckman, J. S., Beckman, T. W., Chen, J., Marshall, P. A., and Freeman, B. A. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 1620-1625
  2. [32]32. Wertz, J. E., and Bolton, J. R. (1972) in Electron Spin Resonance: Elementary Theory and Applications, pp. 450-457, McGraw-Hill Book Co., New York
  3. [38]38. Bisby, R. H., Johnson, S. A., Parker, A. W., and Tavender, S. M. (1998) J. Chem. Soc. Faraday Trans. 94, 2069-2072
  4. [43]43. Gomes, L. F. (1996) The Role of Carbon-centered Radicals on Hydrazine Toxicity. Ph.D. thesis, Instituto de Quı ´mica, Universidade de Sa ˜o Paulo

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Authors
Bonini, M. G., Radi, R., Ferrer-Sueta, G., Ferreira, A. M. D. C., & Augusto, O.
Affiliations
Universidade de São Paulo (BR)
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1999-04
Volume
274
Issue
16
Pages
10802-10806
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2039241150
Cited by
293
References
39
Citations by year
2026: 3 · 2025: 7 · 2024: 6 · 2023: 3 · 2022: 5 · 2021: 4 · 2020: 9 · 2019: 10 · 2018: 9 · 2017: 8 · 2016: 8 · 2015: 6 · 2014: 8 · 2013: 8 · 2012: 13
FWCI
14.68
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Electron Spin Resonance Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
  • Physical Sciences › Chemistry › Organic Chemistry › Free Radicals and Antioxidants
Keywords
carbonate radical anion, peroxynitrite, carbon dioxide, bicarbonate, pH dependence, physiological pH, radical detection
MeSH terms
Carbon Dioxide, Carbonates, Electron Spin Resonance Spectroscopy, Free Radicals, Hydrogen-Ion Concentration, Nitrates

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