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. 1997 Dec 1;328(Pt 2):517–524. doi: 10.1042/bj3280517

Formation of the NO donors glyceryl mononitrate and glyceryl mononitrite from the reaction of peroxynitrite with glycerol.

C R White 1, D Moellering 1, R P Patel 1, M Kirk 1, S Barnes 1, V M Darley-Usmar 1
PMCID: PMC1218950  PMID: 9371710

Abstract

Peroxynitrite (ONOO-), formed from the rapid reaction of superoxide (O2-.) with NO, is known to generate stable compounds capable of donating NO on reaction with thiols and molecules containing hydroxy groups. Using glycerol as a model compound for the reactions of ONOO- with biomolecules containing hydroxy groups, we separated the products and identified them by HPLC/MS. It was shown that both glyceryl mononitrate and glyceryl mononitrite were formed and released NO on incubation with copper and l-cysteine. The compounds were stable over a period of 4h when shielded from light and kept on ice. Slow spontaneous decomposition occurred in the buffer used for the bioassay, but this was not sufficient to explain the vasorelaxing properties of these NO donors. It is concluded that the stable organic nitrate and nitrite have the capacity to be metabolized by vascular tissues, resulting in vasorelaxation.

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Selected References

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  1. Beckman J. S., Beckman T. W., Chen J., Marshall P. A., Freeman B. A. Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1620–1624. doi: 10.1073/pnas.87.4.1620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beckmann J. S., Ye Y. Z., Anderson P. G., Chen J., Accavitti M. A., Tarpey M. M., White C. R. Extensive nitration of protein tyrosines in human atherosclerosis detected by immunohistochemistry. Biol Chem Hoppe Seyler. 1994 Feb;375(2):81–88. doi: 10.1515/bchm3.1994.375.2.81. [DOI] [PubMed] [Google Scholar]
  3. Bennett B. M., McDonald B. J., Nigam R., Simon W. C. Biotransformation of organic nitrates and vascular smooth muscle cell function. Trends Pharmacol Sci. 1994 Jul;15(7):245–249. doi: 10.1016/0165-6147(94)90319-0. [DOI] [PubMed] [Google Scholar]
  4. Buttery L. D., Springall D. R., Chester A. H., Evans T. J., Standfield E. N., Parums D. V., Yacoub M. H., Polak J. M. Inducible nitric oxide synthase is present within human atherosclerotic lesions and promotes the formation and activity of peroxynitrite. Lab Invest. 1996 Jul;75(1):77–85. [PubMed] [Google Scholar]
  5. Chong S., Fung H. L. Biochemical and pharmacological interactions between nitroglycerin and thiols. Effects of thiol structure on nitric oxide generation and tolerance reversal. Biochem Pharmacol. 1991 Sep 12;42(7):1433–1439. doi: 10.1016/0006-2952(91)90456-f. [DOI] [PubMed] [Google Scholar]
  6. Chong S., Fung H. L. Kinetic mechanisms for the concentration dependency of in vitro degradation of nitroglycerin and glyceryl dinitrates in human blood: metabolite inhibition or cosubstrate depletion? J Pharm Sci. 1989 Apr;78(4):295–302. doi: 10.1002/jps.2600780407. [DOI] [PubMed] [Google Scholar]
  7. Darley-Usmar V., Halliwell B. Blood radicals: reactive nitrogen species, reactive oxygen species, transition metal ions, and the vascular system. Pharm Res. 1996 May;13(5):649–662. doi: 10.1023/a:1016079012214. [DOI] [PubMed] [Google Scholar]
  8. Feelisch M., Noack E. A. Correlation between nitric oxide formation during degradation of organic nitrates and activation of guanylate cyclase. Eur J Pharmacol. 1987 Jul 2;139(1):19–30. doi: 10.1016/0014-2999(87)90493-6. [DOI] [PubMed] [Google Scholar]
  9. Fung H. L. Clinical pharmacology of organic nitrates. Am J Cardiol. 1993 Sep 9;72(8):9C–15C. doi: 10.1016/0002-9149(93)90249-c. [DOI] [PubMed] [Google Scholar]
  10. Graham A., Hogg N., Kalyanaraman B., O'Leary V., Darley-Usmar V., Moncada S. Peroxynitrite modification of low-density lipoprotein leads to recognition by the macrophage scavenger receptor. FEBS Lett. 1993 Sep 13;330(2):181–185. doi: 10.1016/0014-5793(93)80269-z. [DOI] [PubMed] [Google Scholar]
  11. Ignarro L. J., Gruetter C. A. Requirement of thiols for activation of coronary arterial guanylate cyclase by glyceryl trinitrate and sodium nitrite: possible involvement of S-nitrosothiols. Biochim Biophys Acta. 1980 Aug 13;631(2):221–231. doi: 10.1016/0304-4165(80)90297-4. [DOI] [PubMed] [Google Scholar]
  12. Ignarro L. J., Lippton H., Edwards J. C., Baricos W. H., Hyman A. L., Kadowitz P. J., Gruetter C. A. Mechanism of vascular smooth muscle relaxation by organic nitrates, nitrites, nitroprusside and nitric oxide: evidence for the involvement of S-nitrosothiols as active intermediates. J Pharmacol Exp Ther. 1981 Sep;218(3):739–749. [PubMed] [Google Scholar]
  13. Ji Y., Akerboom T. P., Sies H. Microsomal formation of S-nitrosoglutathione from organic nitrites: possible role of membrane-bound glutathione transferase. Biochem J. 1996 Jan 15;313(Pt 2):377–380. doi: 10.1042/bj3130377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Knowles R. G., Moncada S. Nitric oxide as a signal in blood vessels. Trends Biochem Sci. 1992 Oct;17(10):399–402. doi: 10.1016/0968-0004(92)90008-w. [DOI] [PubMed] [Google Scholar]
  15. Kowaluk E. A., Fung H. L. Vascular nitric oxide-generating activities for organic nitrites and organic nitrates are distinct. J Pharmacol Exp Ther. 1991 Nov;259(2):519–525. [PubMed] [Google Scholar]
  16. Liu S., Beckman J. S., Ku D. D. Peroxynitrite, a product of superoxide and nitric oxide, produces coronary vasorelaxation in dogs. J Pharmacol Exp Ther. 1994 Mar;268(3):1114–1121. [PubMed] [Google Scholar]
  17. Lymar S. V., Hurst J. K. Carbon dioxide: physiological catalyst for peroxynitrite-mediated cellular damage or cellular protectant? Chem Res Toxicol. 1996 Jul-Aug;9(5):845–850. doi: 10.1021/tx960046z. [DOI] [PubMed] [Google Scholar]
  18. Mayer B., Schrammel A., Klatt P., Koesling D., Schmidt K. Peroxynitrite-induced accumulation of cyclic GMP in endothelial cells and stimulation of purified soluble guanylyl cyclase. Dependence on glutathione and possible role of S-nitrosation. J Biol Chem. 1995 Jul 21;270(29):17355–17360. doi: 10.1074/jbc.270.29.17355. [DOI] [PubMed] [Google Scholar]
  19. Minor R. L., Jr, Myers P. R., Guerra R., Jr, Bates J. N., Harrison D. G. Diet-induced atherosclerosis increases the release of nitrogen oxides from rabbit aorta. J Clin Invest. 1990 Dec;86(6):2109–2116. doi: 10.1172/JCI114949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mohazzab K. M., Wolin M. S. Properties of a superoxide anion-generating microsomal NADH oxidoreductase, a potential pulmonary artery PO2 sensor. Am J Physiol. 1994 Dec;267(6 Pt 1):L823–L831. doi: 10.1152/ajplung.1994.267.6.L823. [DOI] [PubMed] [Google Scholar]
  21. Moncada S., Palmer R. M., Higgs E. A. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev. 1991 Jun;43(2):109–142. [PubMed] [Google Scholar]
  22. Moro M. A., Darley-Usmar V. M., Goodwin D. A., Read N. G., Zamora-Pino R., Feelisch M., Radomski M. W., Moncada S. Paradoxical fate and biological action of peroxynitrite on human platelets. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6702–6706. doi: 10.1073/pnas.91.14.6702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Moro M. A., Darley-Usmar V. M., Lizasoain I., Su Y., Knowles R. G., Radomski M. W., Moncada S. The formation of nitric oxide donors from peroxynitrite. Br J Pharmacol. 1995 Oct;116(3):1999–2004. doi: 10.1111/j.1476-5381.1995.tb16404.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mügge A., Elwell J. H., Peterson T. E., Hofmeyer T. G., Heistad D. D., Harrison D. G. Chronic treatment with polyethylene-glycolated superoxide dismutase partially restores endothelium-dependent vascular relaxations in cholesterol-fed rabbits. Circ Res. 1991 Nov;69(5):1293–1300. doi: 10.1161/01.res.69.5.1293. [DOI] [PubMed] [Google Scholar]
  25. Münzel T., Kurz S., Heitzer T., Harrison D. G. New insights into mechanisms underlying nitrate tolerance. Am J Cardiol. 1996 May 30;77(13):24C–30C. doi: 10.1016/s0002-9149(96)00185-3. [DOI] [PubMed] [Google Scholar]
  26. Münzel T., Sayegh H., Freeman B. A., Tarpey M. M., Harrison D. G. Evidence for enhanced vascular superoxide anion production in nitrate tolerance. A novel mechanism underlying tolerance and cross-tolerance. J Clin Invest. 1995 Jan;95(1):187–194. doi: 10.1172/JCI117637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ohara Y., Peterson T. E., Harrison D. G. Hypercholesterolemia increases endothelial superoxide anion production. J Clin Invest. 1993 Jun;91(6):2546–2551. doi: 10.1172/JCI116491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pritchard K. A., Jr, Groszek L., Smalley D. M., Sessa W. C., Wu M., Villalon P., Wolin M. S., Stemerman M. B. Native low-density lipoprotein increases endothelial cell nitric oxide synthase generation of superoxide anion. Circ Res. 1995 Sep;77(3):510–518. doi: 10.1161/01.res.77.3.510. [DOI] [PubMed] [Google Scholar]
  29. Rajagopalan S., Kurz S., Münzel T., Tarpey M., Freeman B. A., Griendling K. K., Harrison D. G. Angiotensin II-mediated hypertension in the rat increases vascular superoxide production via membrane NADH/NADPH oxidase activation. Contribution to alterations of vasomotor tone. J Clin Invest. 1996 Apr 15;97(8):1916–1923. doi: 10.1172/JCI118623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tarpey M. M., Beckman J. S., Ischiropoulos H., Gore J. Z., Brock T. A. Peroxynitrite stimulates vascular smooth muscle cell cyclic GMP synthesis. FEBS Lett. 1995 May 15;364(3):314–318. doi: 10.1016/0014-5793(95)00413-4. [DOI] [PubMed] [Google Scholar]
  31. Villa L. M., Salas E., Darley-Usmar V. M., Radomski M. W., Moncada S. Peroxynitrite induces both vasodilatation and impaired vascular relaxation in the isolated perfused rat heart. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12383–12387. doi: 10.1073/pnas.91.26.12383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. White C. R., Brock T. A., Chang L. Y., Crapo J., Briscoe P., Ku D., Bradley W. A., Gianturco S. H., Gore J., Freeman B. A. Superoxide and peroxynitrite in atherosclerosis. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):1044–1048. doi: 10.1073/pnas.91.3.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. White C. R., Darley-Usmar V., Berrington W. R., McAdams M., Gore J. Z., Thompson J. A., Parks D. A., Tarpey M. M., Freeman B. A. Circulating plasma xanthine oxidase contributes to vascular dysfunction in hypercholesterolemic rabbits. Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8745–8749. doi: 10.1073/pnas.93.16.8745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wu M., Pritchard K. A., Jr, Kaminski P. M., Fayngersh R. P., Hintze T. H., Wolin M. S. Involvement of nitric oxide and nitrosothiols in relaxation of pulmonary arteries to peroxynitrite. Am J Physiol. 1994 May;266(5 Pt 2):H2108–H2113. doi: 10.1152/ajpheart.1994.266.5.H2108. [DOI] [PubMed] [Google Scholar]

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