Abstract
The peroxidase-catalyzed nitration of tyrosine derivatives by nitrite and hydrogen peroxide has been studied in detail using the enzymes lactoperoxidase (LPO) from bovine milk and horseradish peroxidase (HRP). The results indicate the existence of two competing pathways, in which the nitrating species is either nitrogen dioxide or peroxynitrite. The first pathway involves one-electron oxidation of nitrite by the classical peroxidase intermediates compound I and compound II, whereas in the second pathway peroxynitrite is generated by reaction between enzyme-bound nitrite and hydrogen peroxide. The two mechanisms can be simultaneously operative, and their relative importance depends on the reagent concentrations. With HRP the peroxynitrite pathway contributes significantly only at relatively high nitrite concentrations, but for LPO this represents the main pathway even at relatively low (pathophysiological) nitrite concentrations and explains the high efficiency of the enzyme in the nitration. Myoglobin and hemoglobin are also active in the nitration of phenolic compounds, albeit with lower efficiency compared with peroxidases. In the case of myoglobin, endogenous nitration of the protein has been shown to occur in the absence of substrate. The main nitration site is the heme, but a small fraction of nitrated Tyr146 residue has been identified upon proteolytic digestion and high-performance liquid chromatography/mass spectrometry analysis of the peptide fragments. Preliminary investigation of the nitration of tryptophan derivatives by the peroxidase/nitrite/hydrogen peroxide systems shows that a complex pattern of isomeric nitration products is produced, and this pattern varies with nitrite concentration. Comparative experiments using chemical nitrating agents indicate that at low nitrite concentrations, the enzymatic nitration produces a regioisomeric mixture of nitrotryptophanyl derivatives resembling that obtained using nitrogen dioxide, whereas at high nitrite concentrations the product pattern resembles that obtained using peroxynitrite.
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- Beckman J. S. Oxidative damage and tyrosine nitration from peroxynitrite. Chem Res Toxicol. 1996 Jul-Aug;9(5):836–844. doi: 10.1021/tx9501445. [DOI] [PubMed] [Google Scholar]
- Burner U., Furtmuller P. G., Kettle A. J., Koppenol W. H., Obinger C. Mechanism of reaction of myeloperoxidase with nitrite. J Biol Chem. 2000 Jul 7;275(27):20597–20601. doi: 10.1074/jbc.M000181200. [DOI] [PubMed] [Google Scholar]
- Casella L., Poli S., Gullotti M., Selvaggini C., Beringhelli T., Marchesini A. The chloroperoxidase-catalyzed oxidation of phenols. Mechanism, selectivity, and characterization of enzyme-substrate complexes. Biochemistry. 1994 May 31;33(21):6377–6386. doi: 10.1021/bi00187a001. [DOI] [PubMed] [Google Scholar]
- Exner M., Herold S. Kinetic and mechanistic studies of the peroxynitrite-mediated oxidation of oxymyoglobin and oxyhemoglobin. Chem Res Toxicol. 2000 Apr;13(4):287–293. doi: 10.1021/tx990201k. [DOI] [PubMed] [Google Scholar]
- Floris R., Piersma S. R., Yang G., Jones P., Wever R. Interaction of myeloperoxidase with peroxynitrite. A comparison with lactoperoxidase, horseradish peroxidase and catalase. Eur J Biochem. 1993 Aug 1;215(3):767–775. doi: 10.1111/j.1432-1033.1993.tb18091.x. [DOI] [PubMed] [Google Scholar]
- Huie R. E., Padmaja S. The reaction of no with superoxide. Free Radic Res Commun. 1993;18(4):195–199. doi: 10.3109/10715769309145868. [DOI] [PubMed] [Google Scholar]
- Ischiropoulos H. Biological tyrosine nitration: a pathophysiological function of nitric oxide and reactive oxygen species. Arch Biochem Biophys. 1998 Aug 1;356(1):1–11. doi: 10.1006/abbi.1998.0755. [DOI] [PubMed] [Google Scholar]
- Koppenol W. H., Moreno J. J., Pryor W. A., Ischiropoulos H., Beckman J. S. Peroxynitrite, a cloaked oxidant formed by nitric oxide and superoxide. Chem Res Toxicol. 1992 Nov-Dec;5(6):834–842. doi: 10.1021/tx00030a017. [DOI] [PubMed] [Google Scholar]
- Monzani E., Alzuet G., Casella L., Redaelli C., Bassani C., Sanangelantoni A. M., Gullotti M., de Gioia L., Santagostini L., Chillemi F. Properties and reactivity of myoglobin reconstituted with chemically modified protohemin complexes. Biochemistry. 2000 Aug 8;39(31):9571–9582. doi: 10.1021/bi000784t. [DOI] [PubMed] [Google Scholar]
- Monzani E., Gatti A. L., Profumo A., Casella L., Gullotti M. Oxidation of phenolic compounds by lactoperoxidase. Evidence for the presence of a low-potential compound II during catalytic turnover. Biochemistry. 1997 Feb 18;36(7):1918–1926. doi: 10.1021/bi961868y. [DOI] [PubMed] [Google Scholar]
- Pfeiffer S., Mayer B. Lack of tyrosine nitration by peroxynitrite generated at physiological pH. J Biol Chem. 1998 Oct 16;273(42):27280–27285. doi: 10.1074/jbc.273.42.27280. [DOI] [PubMed] [Google Scholar]
- Sampson J. B., Ye Y., Rosen H., Beckman J. S. Myeloperoxidase and horseradish peroxidase catalyze tyrosine nitration in proteins from nitrite and hydrogen peroxide. Arch Biochem Biophys. 1998 Aug 15;356(2):207–213. doi: 10.1006/abbi.1998.0772. [DOI] [PubMed] [Google Scholar]
- Wu W., Chen Y., Hazen S. L. Eosinophil peroxidase nitrates protein tyrosyl residues. Implications for oxidative damage by nitrating intermediates in eosinophilic inflammatory disorders. J Biol Chem. 1999 Sep 3;274(36):25933–25944. doi: 10.1074/jbc.274.36.25933. [DOI] [PubMed] [Google Scholar]
- Ye Y. Z., Strong M., Huang Z. Q., Beckman J. S. Antibodies that recognize nitrotyrosine. Methods Enzymol. 1996;269:201–209. doi: 10.1016/s0076-6879(96)69022-3. [DOI] [PubMed] [Google Scholar]
- van der Vliet A., Eiserich J. P., Halliwell B., Cross C. E. Formation of reactive nitrogen species during peroxidase-catalyzed oxidation of nitrite. A potential additional mechanism of nitric oxide-dependent toxicity. J Biol Chem. 1997 Mar 21;272(12):7617–7625. doi: 10.1074/jbc.272.12.7617. [DOI] [PubMed] [Google Scholar]