Role of the carbonate radical anion in tyrosine nitration and hydroxylation by peroxynitrite

Célio X.C. Santos, Marcelo G. Bonini, Ohara Augusto*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

103 Scopus citations

Abstract

Peroxynitrite has been receiving increasing attention as the pathogenic mediator of nitric oxide cytotoxicity. In most cases, the contribution of peroxynitrite to diseases has been inferred from detection of 3-nitrotyrosine in injured tissues. However, presently it is known that other nitric oxide- derived species can also promote protein nitration. Mechanistic details of protein nitration remain under discussion even in the case of peroxynitrite, although recent literature data strongly suggest a free radical mechanism. Here, we confirm the free radical mechanism of tyrosine modification by peroxynitrite in the presence and in the absence of the bicarbonate-carbon dioxide pair by analyzing the stable tyrosine products and the formation of the tyrosyl radical at pH 5.4 and 7.4. Stable products, 3-nitrotyrosine, 3- hydroxytyrosine, and 3,3-dityrosine, were identified by high performance liquid chromatography and UV spectroscopy. The tyrosyl radical was detected by continuous-flow and spin-trapping electron paramagnetic resonance (EPR). 3-Hydroxytyrosine was detected at pH 5.4 and its yield decreased in the presence of the bicarbonate-carbon dioxide pair. In contrast, the yields of the tyrosyl radical increased in the presence of the bicarbonate-carbon dioxide pair and correlated with the yields of 3-nitrotyrosine under all tested experimental conditions. Taken together, the results demonstrate that the promoting effects of carbon dioxide on peroxynitrite-mediated tyrosine nitration is due to the selective reactivity of the carbonate radical anion as compared with that of the hydroxyl radical. Colocalization of 3- hydroxytyrosine and 3-nitrotyrosine residues in proteins may be useful to discriminate between peroxynitrite and other nitrating species. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)146-152
Number of pages7
JournalArchives of biochemistry and biophysics
Volume377
Issue number1
DOIs
StatePublished - May 1 2000
Externally publishedYes

Keywords

  • Carbonate radical anion
  • DOPA
  • Nitric oxide
  • Nitrotyrosine
  • Peroxynitrite
  • Tyrosyl radical

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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