Nitric oxide (˙NO) plays a central role in the pathogenesis of diverse inflammatory and infectious disorders1,2. The toxicity of ˙NO is thought to be engendered, in part, by its reaction with superoxide (O˙−2), yielding the potent oxidant peroxynitrite (ONOO−)3. However, evidence for a role of ONOO− in vivo is based largely upon detection of 3-nitrotyrosine in injured tissues4,5,6,7,8. We have recently demonstrated that nitrite (NO2−), a major end-product of ˙NO metabolism, readily promotes tyrosine nitration through formation of nitryl chloride (NO2Cl) and nitrogen dioxide (˙NO2) by reaction with the inflammatory mediators hypochlorous acid (HOCl) or myeloperoxidase9,10. We now show that activated human polymorphonuclear neutrophils convert NO2− into NO2Cl and ˙NO2 through myeloperoxidase-dependent pathways. Polymorphonuclear neutrophil-mediated nitration and chlorination of tyrosine residues or 4-hydroxyphenylacetic acid is enhanced by addition of NO2− or by fluxes of ˙NO. Addition of 15NO2− led to 15N enrichment of nitrated phenolic substrates, confirming its role in polymorphonuclear neutrophil-mediated nitration reactions. Polymorphonuclear neutrophil-mediated inactivation of endothelial cell angiotensin-converting enzyme was exacerbated by NO2−, illustrating the physiological significance of these reaction pathways to cellular dysfunction. Our data reveal that NO2− may regulate inflammatory processes through oxidative mechanisms, perhaps by contributing to the tyrosine nitration and chlorination observed in vivo.
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We thank M. Shigenaga, S. Christen, A. Estevez, R. Radi and B. Alvarez for helpful comments and discussions, S. E. Ebeler and M. R. Webb for assistance with mass spectrometry, and J.Catravas and J. Ryan for kindly providing [3H]BPAP. This work was supported by grants from the NIH, the Cystic Fibrosis Foundation, and the Arthritis and Rheumatism Council (UK). J.P.E. is a recipient of a Research Fellowship from the California Affiliate of the American Lung Association and A.v.d.V. is a Parker B. Francis Fellow in Pulmonary Research.
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