Preferences help
enabled [disable] Abstract
Number of results
2000 | 47 | 4 | 913-921
Article title

Mitochondria recycle nitrite back to the bioregulator nitric monoxide.

Title variants
Languages of publication
Nitric monoxide (NO) exerts a great variety of physiological functions. L-Arginine supplies amino groups which are transformed to NO in various NO-synthase-active isoenzyme complexes. NO-synthesis is stimulated under various conditions increasing the tissue of stable NO-metabolites. The major oxidation product found is nitrite. Elevated nitrite levels were reported to exist in a variety of diseases including HIV, reperfusion injury and hypovolemic shock. Denitrifying bacteria such as Paracoccus denitrificans have a membrane bound set of cytochromes (cyt cd1, cyt bc) which were shown to be involved in nitrite reduction activities. Mammalian mitochondria have similar cytochromes which form part of the respiratory chain. Like in bacteria quinols are used as reductants of these types of cytochromes. The observation of one-e- divergence from this redox-couple to external dioxygen made us to study whether this site of the respiratory chain may also recycle nitrite back to its bioactive form NO. Thus, the aim of the present study was therefore to confirm the existence of a reductive pathway which reestablishes the existence of the bioregulator NO from its main metabolite NO2-. Our results show that respiring mitochondria readily reduce added nitrite to NO which was made visible by nitrosylation of deoxyhemoglobin. The adduct gives characteristic triplet-ESR-signals. Using inhibitors of the respiratory chain for chemical sequestration of respiratory segments we were able to identify the site where nitrite is reduced. The results confirm the ubiquinone/cyt bc1 couple as the reductant site where nitrite is recycled. The high affinity of NO to the heme-iron of cytochrome oxidase will result in an impairment of mitochondrial energy-production. "Nitrite tolerance" of angina pectoris patients using NO-donors may be explained in that way.

Physical description
  • Institute of Pharmacology and Toxicology, University of Veterinary Medicine, Vienna, Austria
  • Institute of Pharmacology and Toxicology, University of Veterinary Medicine, Vienna, Austria
  • Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Vienna, Austria
  • Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Vienna, Austria
  • Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Vienna, Austria
  • Institute of Pharmacology and Toxicology, University of Veterinary Medicine, Vienna, Austria
  • 1. Palmer, R.M., Ferrige, A.G. & Moncada, S. (1987) Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature 327, 524-526.
  • 2. Henry, Y., Lepoivre, M., Drapier, J.C., Ducrocq, C., Boucher, J.L. & Guissani, A. (1993) EPR characterization of molecular targets for NO in mammalian cells and organelles. FASEB J. 7, 1124-1134.
  • 3. Seligman, S.P., Buyon, J.P., Clancy, R.M., Young, B.K. & Abramson, S.B. (1994) The role of nitric oxide in the pathogenesis of preeclampsia. Am. J. Obstet. Gynecol. 171, 944- 948.
  • 4. Tsikas, D., Boger, R.H., Bode Boger, S.M., Gutzki, F.M. & Frolich, J.C. (1994) Quantification of nitrite and nitrate in human urine and plasma as pentafluorobenzyl derivatives by gas chromatography-mass spectrometry using their 15N-labelled analogs. J. Chromatogr. B. Biomed. Appl. 661, 185-191.
  • 5. Preik Steinhoff, H. & Kelm, M. (1996) Determination of nitrite in human blood by combination of a specific sample preparation with high-performance anion-exchange chromatography and electrochemical detection. J. Chromatogr. B. Biomed. Appl. 685, 348-352.
  • 6. Komori, K., Matsumoto, T., Ishida, M., Kuma, S., Yonemitsu, Y., Eguchi, D. & Sugimachi, K. (1997) Enhancement of nitric oxide production after arterial reconstruction in patients with arteriosclerosis obliterans. J. Vasc. Surg. 26, 657-662.
  • 7. Zweier, J.L., Wang, P., Samouilov, A. & Kuppusamy, P. (1995) Enzyme-independent formation of nitric oxide in biological tissues. Nat. Med. 1, 804-809.
  • 8. Vanin, A.F., Kiladze, S.V. & Kubrina, L.N. (1977) Factors influencing formation of dinitrosyl complexes of non-heme iron in the organs of animals in vivo. Biofizika 22, 850-855.
  • 9. Zhang, Z., Naughton, D., Winyard, P.G., Benjamin, N., Blake, D.R. & Symons, M.C.R. (1998) Generation of nitric oxide by a nitrite reductase activity of xanthine oxidase: A potential pathway for nitric oxide formation in the absence of nitric oxide synthase activity [published erratum appears in Biochem Biophys. Res. Commun. (1998) Biochem. Biophys. Res. Commun. 249, 767- 772.
  • 10. Stolze, K., Dadak, A., Liu, Y. & Nohl, H. (1996) Hydroxylamine and phenol-induced formation of methemoglobin and free radical intermediates in erythrocytes. Biochem. Pharmacol. 52, 1821-1829.
  • 11. Szarkowska, L. & Klingenberg, M. (1963) On the role of ubiquinone in mitochondria, Spectrophotometric and chemical measurements of its redox reaction. Biochem. Z. 338, 674- 697.
  • 12. Graham, J.M. & Rickwood, D. (1997) Subcellular Fractionation. Oxford University Press, Oxford, New York, Tokyo.
  • 13. Kozlov, A.V., Bini, A., Iannone, A., Zini, I. & Tomasi, A. (1996) Electron paramagnetic resonance characterization of rat neuronal nitric oxide production ex vivo. Methods Enzymol. 268, 229-236.
  • 14. Kozlov, A.V., Sobhian, B., Duvigneau, C., Costantino, G., Gemeiner, M., Nohl, H., Redl, H. & Bahrami, S. (1999) NO synthase-dependent/independent formation of NO after intestinal ischemia and reperfusion. Shock (Suppl.) 12, 44.
  • 15. Kozlov, A.V., Sobhian, B., Duvigneau, C., Gemeiner, M., Nohl, H., Redl, H. & Bahrami, S. (2000) Organ specific formation of nitrosyl complexes under intestinal ischemia/reperfusion in rats involves NOS-independent mechanism(s). Shock, in press.
  • 16. Henry, Y. & Bessieres, P. (1984) Denitrification and nitrite reduction: Pseudomonas aeruginosa nitrite-reductase. Biochimie 66, 259-289.
  • 17. Carr, G.J., Page, M.D. & Ferguson, S.J. (1989) The energy-conserving nitric-oxide-reductase system in Paracoccus denitrificans. Distinction from the nitrite reductase that catalyses synthesis of nitric oxide and evidence from trapping experiments for nitric oxide as a free intermediate during denitrification. Eur. J. Biochem. 179, 683-692.
  • 18. Bessieres, P. & Henry, Y. (1984) Stoichiometry of nitrite reduction catalyzed by Pseudomonas aeruginosa nitrite-reductase. Biochimie 66, 313-318.
  • 19. Zweier, J.L., Samouilov, A. & Kuppusamy, P. (1999) Non-enzymatic nitric oxide synthesis in biological systems. Biochim. Biophys. Acta, 1411, 250-262.
  • 20. Ghafourifar, P. & Richter, C. (1997) Nitric oxide synthase activity in mitochondria. FEBS Lett. 418, 291-296.
  • 21. Giulivi, C., Poderoso, J.J. & Boveris, A. (1998) Production of nitric oxide by mitochondria. J. Biol. Chem. 273, 11038-11043.
  • 22. Nohl, H., Gille, L. & Kozlov, A.V. (1998) Prooxidant functions of coenzyme Q. Subcell. Biochem. 30, 509-526.
  • 23. Brown, G.C. (1995) Nitric oxide regulates mitochondrial respiration and cell functions by inhibiting cytochrome oxidase. FEBS Lett. 369, 136-139.
  • 24. Ghafourifar, P., Schenk, U., Klein, S.D. & Richter, C. (1999) Mitochondrial nitric-oxide synthase stimulation causes cytochrome c release from isolated mitochondria. Evidence for intramitochondrial peroxynitrite formation. J. Biol. Chem. 274, 31185-31188.
  • 25. Boveris, A., Costa, L.E., Poderoso, J.J., Carreras, M.C. & Cadenas, E. (2000) Regulation of mitochondrial respiration by oxygen and nitric oxide. Ann. N.Y. Acad. Sci. U.S.A. 899, 121-135.
  • 26. Takehara, Y., Nakahara, H., Inai, Y., Yabuki, M., Hamazaki, K., Yoshioka, T., Inoue, M., Horton, A.A. & Utsumi, K. (1996) Oxygen-dependent reversible inhibition of mitochondrial respiration by nitric oxide. Cell Struct. Funct. 21, 251-258.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.