The peroxynitrite anion is a potent oxidizing agent, formed by the diffusion-limited combination of nitric oxide and superoxide, and its production under physiological conditions is associated with the pathologies of a number of inflammatory and neurodegenerative diseases. Nitration of Escherichia coli iron superoxide dismutase (Fe-SOD) by peroxynitrite was investigated, and demonstrated by spectral changes and electrospray mass spectroscopic analysis. HPLC and mass studies of the tryptic digests of the mono-nitrated Fe-SOD indicated that tyrosine-34 was the residue most susceptible to nitration by peroxynitrite. Exclusive nitration of this residue occurred when Fe-SOD was exposed to a cumulative dose of 0.4mM peroxynitrite. Unlike with human Mn-SOD, this single modification did not inactivate E. coli Fe-SOD at pH7.4. When Fe-SOD was exposed to higher concentrations of peroxynitrite (7mM), eight tyrosine residues per subunit of the protein, of the nine available, were nitrated without loss of catalytic activity of the enzyme. The pKa of nitrated tyrosine-34 was determined to be 7.95±0.15, indicating that the peroxynitrite-modified enzyme appreciably maintains its protonation state under physiological conditions.
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December 2001
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Research Article|
December 10 2001
Peroxynitrite-induced nitration of tyrosine-34 does not inhibit Escherichia coli iron superoxide dismutase
Laurent SOULÈRE;
Laurent SOULÈRE
Groupe de Chimie Organique Biologique, Laboratoire de Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique, UMR 5068, Université Paul Sabatier, 118 route de Narbonne, 31062 Toulouse cedex 4, France
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Catherine CLAPAROLS;
Catherine CLAPAROLS
Groupe de Chimie Organique Biologique, Laboratoire de Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique, UMR 5068, Université Paul Sabatier, 118 route de Narbonne, 31062 Toulouse cedex 4, France
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Jacques PÉRIÉ;
Jacques PÉRIÉ
Groupe de Chimie Organique Biologique, Laboratoire de Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique, UMR 5068, Université Paul Sabatier, 118 route de Narbonne, 31062 Toulouse cedex 4, France
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Pascal HOFFMANN
Pascal HOFFMANN
1
Groupe de Chimie Organique Biologique, Laboratoire de Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique, UMR 5068, Université Paul Sabatier, 118 route de Narbonne, 31062 Toulouse cedex 4, France
1To whom correspondence should be addressed (e-mail hoffmann@cict.fr).
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Publisher: Portland Press Ltd
Received:
June 29 2001
Revision Received:
September 17 2001
Accepted:
October 15 2001
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2001
2001
Biochem J (2001) 360 (3): 563–567.
Article history
Received:
June 29 2001
Revision Received:
September 17 2001
Accepted:
October 15 2001
Citation
Laurent SOULÈRE, Catherine CLAPAROLS, Jacques PÉRIÉ, Pascal HOFFMANN; Peroxynitrite-induced nitration of tyrosine-34 does not inhibit Escherichia coli iron superoxide dismutase. Biochem J 15 December 2001; 360 (3): 563–567. doi: https://doi.org/10.1042/bj3600563
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