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Formation of nitric oxide-derived inflammatory oxidants by myeloperoxidase in neutrophils

Nature volume 391, pages 393–397 (1998)Cite this article

Abstract

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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Figure 1: Nitrite enhances PMN-mediated chlorination and nitration of phenolic substrates by MPO-dependent mechanisms.
Figure 2: Nitrite competes with taurine for reaction with PMN-released HOCl.
Figure 3: Dependence of NO2− on reaction pathways mediated by PMN exposed to ˙NO.
Figure 4: Nitrite exacerbates PMN-mediated inactivation of angiotensin-converting enzyme (ACE) in bovine aortic endothelial cells (BAEC).

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References

  1. Moncada, S. & Higgs, A. The L-arginine-nitric oxide pathway. N. Engl. J. Med. 329, 2002–2012 (1993).

    Article  CAS  Google Scholar 

  2. Wei, X.et al. Altered immune responses in mice lacking inducible nitric oxide synthase. Nature 375, 408–411 (1995).

    Article  ADS  CAS  Google Scholar 

  3. Beckman, J. S. Oxidative damage and tyrosine nitration from peroxynitrite. Chem. Res. Toxicol. 9, 836–844 (1996).

    Article  CAS  Google Scholar 

  4. Beckman, J. S.et al. Extensive nitration of protein tyrosines in human atherosclerosis detected by immunohistochemistry. Biol. Chem. Hoppe-Seyler 375, 81–88 (1994).

    Article  Google Scholar 

  5. Kaur, H. & Halliwell, B. Evidence for nitric oxide-mediated oxidative damage in chronic inflammation. Nitrotyrosine in serum and synovial fluid from rheumatoid patients. FEBS Lett. 350, 9–12 (1994).

    Article  CAS  Google Scholar 

  6. Kooy, N. W., Royall, J. A., Ye, Y. Z., Kelly, D. R. & Beckman, J. S. Evidence for in vivo peroxynitrite production in human acute lung injury. Am. J. Respir. Crit. Care Med. 151, 1250–1254 (1995).

    CAS  PubMed  Google Scholar 

  7. Singer, I. I.et al. Expression of inducible nitric oxide synthease and nitrotyrosine in colonic epithelium in inflammatory bowel disease. Gastroenterology 111, 871–885 (1996).

    Article  CAS  Google Scholar 

  8. Saleh, D., Barnes, P. J. & Giaid, A. Increased production of the potent oxidant peroxynitrite in the lungs of patients with idiopathic pulmonary fibrosis. Am. J. Respir. Crit. Care Med. 155, 1763–1769 (1997).

    Article  CAS  Google Scholar 

  9. Eiserich, J. P., Cross, C. E., Jones, A. D., Halliwell, B. & van der Vliet, A. Formation of nitrating and chlorinating species by reaction of nitrite with hypochlorous acid. A novel mechanism for nitric oxide-mediated protein modification. J. Biol. Chem. 271, 19199–19208 (1996).

    Article  CAS  Google Scholar 

  10. 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. 272, 7617–7625 (1997).

    Article  CAS  Google Scholar 

  11. Klebanoff, S. J. & Clark, R. A. in The Neutrophil: Function and Clinical Disorders (North Holland Publishing, Amsterdam, 1978).

    Google Scholar 

  12. Weiss, S. J. Tissue destruction by neutrophils. N. Engl. J. Med. 320, 365–376 (1989).

    Article  CAS  Google Scholar 

  13. Hazen, S. L., Hsu, F. F., Mueller, D. M., Crowley, J. R. & Heinecke, J. W. Human neutrophils employ chlorine gas as an oxidant during phagocytosis. J. Clin. Invest. 98, 1283–1289 (1996).

    Article  CAS  Google Scholar 

  14. Weiss, S. J., Lampert, M. B. & Test, S. T. Long-lived oxidants generated by human neutrophils. Characterization and bioactivity. Science 222, 625–628 (1983).

    Article  ADS  CAS  Google Scholar 

  15. Domigan, N. M., Charlton, T. S., Duncan, M. W., Winterbourn, C. C. & Kettle, A. J. Chlorination of tyrosyl residues in peptides by myeloperoxidase and human neutrophils. J. Biol. Chem. 270, 16542–16548 (1995).

    Article  CAS  Google Scholar 

  16. Farrell, A. J., Blake, D. R., Palmer, R. M. J. & Moncada, S. Increased concentrations of nitrite in synovial fluid and serum samples suggest increased nitric oxide synthesis in rheumatic diseases. Ann. Rheum. Dis. 51, 1219–1222 (1992).

    Article  CAS  Google Scholar 

  17. Torre, D.et al. Serum concentrations of nitrite in patients with HIV-1 infection. J. Clin. Pathol. 49, 574–576 (1996).

    Article  CAS  Google Scholar 

  18. Gaston, B.et al. Endogenous nitrogen oxides and bronchodilator S-nitrosothiols in human airways. Proc. Natl Acad. Sci. USA 90, 10957–10961 (1993).

    Article  ADS  CAS  Google Scholar 

  19. Kettle, A. J., Gedye, C. A., Hampton, M. B. & 1Winterbourn, C. C. Inhibition of myeloperoxidase by benzoic acid hydrazides. Biochem. J. 308, 559–563 (1995).

    Article  CAS  Google Scholar 

  20. Heinecke, J. W., Li, W., Daehnke, H. L. & Goldstein, J. A. Dityrosine, a specific marker of oxidation, is synthesized by the myeloperoxidase-hydrogen peroxide system of human neutrophils and macrophages. J. Biol. Chem. 268, 4069–4077 (1993).

    CAS  PubMed  Google Scholar 

  21. Prütz, W. A., Mönig, H., Butler, J. & Land, E. J. Reactions of nitrogen dioxide in aqueous model systems. Oxidation of tyrosine units in peptides and proteins. Arch. Biochem. Biophys. 243, 125–134 (1985).

    Article  Google Scholar 

  22. Mooradian, D. L., Hutsell, T. C. & Keefer, L. K. Nitric oxide (NO) donor molecules. Effect of NO release rate on vascular smooth muscle cell proliferation in vitro. J. Cardiovasc. Pharmacol. 25, 674–678 (1995).

    Article  CAS  Google Scholar 

  23. Chen, X. & Catravas, J. D. Neutrophil-mediated endothelial angiotensin-converting enzyme dysfunction. Role of oxygen-derived free radicals. Am. J. Physiol. 265, L243–L249 (1993).

    CAS  PubMed  Google Scholar 

  24. Corvol, P., Williams, T. A. & Soubrier, F. Peptidyl dipeptidase A: angiotensin I-converting enzyme. Methods Enzymol. 248, 283–305 (1995).

    Article  CAS  Google Scholar 

  25. Daugherty, A., Dunn, J. L., Rateri, D. L. & Heinecke, J. W. Myeloperoxidase, a catalyst for lipoprotein oxidation, is expressed in human atherosclerotic lesions. J. Clin. Invest. 94, 437–444 (1994).

    Article  CAS  Google Scholar 

  26. Hazen, S. L. & Heinecke, J. W. 3-Chlorotyrosine, a specific marker of myeloperoxidase-catalyzed oxidation, is markedly elevated in low density lipoprotein isolated from human atherosclerotic intima. J. Clin. Invest. 99, 2075–2081 (1997).

    Article  CAS  Google Scholar 

  27. Evans, T. J.et al. Cytokine-treated human neutrophils contain inducible nitric oxide synthase that produces nitration of ingested bacteria. Proc. Natl Acad. Sci. USA 93, 9553–9558 (1996).

    Article  ADS  CAS  Google Scholar 

  28. Klebanoff, S. J. Reactive nitrogen intermediates and antimicrobial activity. Role of nitrite. Free Rad. Biol. Med. 14, 351–360 (1993).

    Article  CAS  Google Scholar 

  29. Gunther, M. R.et al. Nitric oxide trapping of the tyrosyl radical of prostaglandin H synthease-2 leads to tyrosine iminoxyl radical and nitrotyrosine formation. J. Biol. Chem. 272, 17086–17090 (1997).

    Article  CAS  Google Scholar 

  30. Simon, D. I.et al. Polynitrosylated proteins. Characterization, bioactivity, and functional consequences. Proc. Natl Acad. Sci. USA 93, 4736–4741 (1996).

    Article  ADS  CAS  Google Scholar 

Download references

Acknowledgements

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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  1. Jason P. Eiserich

    Present address: Department of Anesthesiology, UAB Center for Free Radical Biology, The University of Alabama at Birmingham, 619 19th Street South, 946 THT, Birmingham, Alabama, 35233, USA

Authors and Affiliations

  1. Department of Internal Medicine, Division of Pulmonary and Critical Care Medicine, University of California, Davis, 95616, California, USA

    Jason P. Eiserich, Milena Hristova, Carroll E. Cross, Barry Halliwell & Albert van der Vliet

  2. Facility for Advanced Instrumentation, University of California, Davis, 95616, California, USA

    A. Daniel Jones

  3. Departments of Anesthesiology, Biochemistry and Molecular Genetics, Pediatrics, and the UAB Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, 35233, Alabama, USA

    Bruce A. Freeman

  4. Neurodegenerative Disease Research Centre, Pharmacology Group, University of London, King's College, SW3 6LX, London, UK

    Barry Halliwell

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Correspondence to Jason P. Eiserich.

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Eiserich, J., Hristova, M., Cross, C. et al. Formation of nitric oxide-derived inflammatory oxidants by myeloperoxidase in neutrophils. Nature 391, 393–397 (1998). https://doi.org/10.1038/34923

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