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

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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

DOI 10.1074/jbc.274.16.10802

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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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Authors: Bonini, M. G., Radi, R., Ferrer-Sueta, G., Ferreira, A. M. D. C., & Augusto, O.
Affiliations: Universidade de São Paulo (BR)
ORCID: Bonini, Marcelo G.
Ferreira, Ana M. Da C.
Venue: Journal of Biological Chemistry
Publisher: Elsevier BV
Published: 1999-04
Volume: 274
Issue: 16
Pages: 10802-10806
ISSN: 0021-9258 (ISSN-L)
DOI: 10.1074/jbc.274.16.10802
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: hybrid
doi.org
Licence: cc-by
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
creativecommons.org/licenses/by/4.0/ (vor, 7857 d)
Locations: Journal of Biological Chemistry (publishedVersion, cc-by)
PubMed (publishedVersion)
Topics: Physical Sciences › Chemistry › Spectroscopy › 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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Direct EPR Detection of the Carbonate Radical Anion Produced from Peroxynitrite and Carbon Dioxide

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