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Direct EPR Detection of the Carbonate Radical Anion Produced from Peroxynitrite and Carbon Dioxide
Open accessAbstract
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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“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).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [112]
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Metadata
- 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
- hybrid
- doi.org
- Licence
- 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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