The Molecular Mechanism of Interaction of H2O2 with Metmyoglobin

  • Dimitrios Galaris
  • Stelianos Kokkoris
  • Ioannis Toumpoulis
  • Panagiotis Korantzopoulos
Part of the NATO ASI Series book series (NSSA, volume 296)


Slight changes in the intacellular redox equilibrium is a physiological situation utilized by nature to regulate many important cell functions (Sundaresan et al., 1995). However, when the level of the oxidizing substances produced exceeds a certain threshold it becomes deleterious for the cells (Halliwell and Gutteridge, 1989). The main source of oxidizing agents is the monovalent reduction of oxygen to superoxide anion and the subsequent formation of reactive oxygen intermediates (ROI) (Figure 1).


Ascorbic Acid Electron Spin Resonance Peroxyl Radical Reactive Oxygen Intermediate Oxidizable Amino Acid 
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  1. Allentoff, A.J., Bolton, J.L., Wilks, A., Thompson, J.A., and Ortiz de Montellano, P.R., 1992, Heterolytic versus homolytic peroxide bond cleavage by sperm whale myoglobin and myoglobin mutants, J. Am. Chem. Soc. 114: 9744–9749.CrossRefGoogle Scholar
  2. Arduini, A., Eddy, L., and Hochstein, P., 1990, Detection of ferrylmyoglobin in the isolated ischemic rat heart, Free Radic. Biol. Med. 9: 511–513.CrossRefGoogle Scholar
  3. Catalano, C.E., Choe, Y.S., and Ortiz de Montellano, P.R., 1989, Reactions of the protein radical in peroxide-treated myoglobin, J. Biol. Chem. 264: 10534–10541.Google Scholar
  4. Chance, B., 1952, The spectra of the enzyme-substrate complexes of catalase and peroxidases, Arch. Biochem. Biophys. 41: 404–415.CrossRefGoogle Scholar
  5. Eddy, L., Arduini, A., and Hochstein, P., 1990a, Reduction of ferrylmyoglobin in rat diaphragm, Am. J. Physiol. 259: C995–C997.Google Scholar
  6. Eddy, L., Hurvitz, R., and Hochstein, P., 1990b, A protective role for ascorbate in ischemic arrest associated with cardiopulmonary bypass, J. Appl. Cardiol. 5: 409–414.Google Scholar
  7. Evans, P.J., and Halliwell, B., 1995, Side-effects of drugs used in the treatment of rheumatoid arthritis, Biochem. Soc. Symp. 61: 195–207.Google Scholar
  8. Galaris D., and Korantzopoulos P., 1997, On the molecular mechanism of metmyoglobin-catalyzed reduction of hydrogen peroxide by ascorbate, Free Radic. Biol. Med. 22: 657–667.CrossRefGoogle Scholar
  9. Galaris, D., Mira, D., Sevanian, A., Cadenas, E., and Hochstein, P., 1988, Co-oxidation of salicylate and cholesterol and generation of electronically-excited states during the oxidation of metmyoglobin by H202, Arch. Biochem. Biophys. 281: 163–169.CrossRefGoogle Scholar
  10. Galaris, D., Cadenas, E., and Hochstein, P., 1989a, Glutathione-dependent reduction of peroxides during ferryland met-myoglobin interconversion: A potential protective mechanism in muscle, Free Radic. Biol. Med. 6: 473–478.CrossRefGoogle Scholar
  11. Galaris, D., Cadenas, E., and Hochstein, P., 19896, Redox-cycling of myoglobin and ascorbate: A potential protective mechanism against oxidative reperfusion injury in muscle, Arch. Biochem. Biophys. 273: 497–504.Google Scholar
  12. Galaris, D., Eddy, L. Arduini, A., Cadenas, E., and Hochstein, P., 1989c, Mechanisms of reoxygenation injury in myocardial infarction: Implication of a myoglobin redox cycle, Biochem. Biophys. Res. Commun. 160: 1162–1168.CrossRefGoogle Scholar
  13. Galaris, D., Kokkoris, S., Toumpoulis., I., and Tsolas, O., 1997, Generation of reactive species and arachidonic acid peroxidation during the oxidation of metmyoglobin by hydrogen peroxide, Submitted for publication.Google Scholar
  14. George, P., and Irvine, D.H., 1952, The reaction between metmyoglobin and hydrogen peroxide, Biochem. J. 52: 511–517.Google Scholar
  15. Giulivi, C., and Cadenas, E., 1993, The reaction of ascorbic acid with different heme iron states of myoglobin, FEBS Lett. 332: 287–290.CrossRefGoogle Scholar
  16. Giulivi, C., and Cadenas, E., 1994, Ferrylmyoglobin: Formation and chemical reactivity toward electron-donating compounds, Meth. Enzymol. 233: 189–202.CrossRefGoogle Scholar
  17. Giulivi, C., Romero, F.J., and Cadenas, E., 1992, The interaction of Trolox C, a water-soluble vitamin E analog with ferrylmyoglobin: Reduction of the oxoferryl moiety, Arch. Biochem. Biophys. 299: 302–312.CrossRefGoogle Scholar
  18. Grisham, M.B., 1985, Myoglobin-catalyzed hydrogen peroxide dependent arachidonic acid peroxidation, J. Free Radic. Biol. Med. 1: 227–232.CrossRefGoogle Scholar
  19. Gunther, M.R., Kelman, D.J., Corbett, J.T., and Mason, R.P., 1995, Self-peroxidation of metmyoglobin results in formation of an oxygen reactive tryptophan-centered radical, J. Biol. Chem. 270: 16075–16081.CrossRefGoogle Scholar
  20. Halliwell, B., and Gutteridge, J.M.C., 1989, Free Radicals in Biology and Medicine, 2nd ed., Clarendon Press, Oxford.Google Scholar
  21. Hanan T., and Shaklai, N., 1995, Peroxidative interaction of myoglobin and myosin, Eur. J. Biochem. 233: 930–936.CrossRefGoogle Scholar
  22. Harel, S., and Kanner, J., 1988, The generation of ferryl or hydroxyl radicals during interaction of haemproteins with hydrogen peroxide, Free Radic. Res. Commun. 5: 21–33.CrossRefGoogle Scholar
  23. Kanner, J., and Harel, S., 1985, Initiation of membranal lipid peroxidation by activated metmyoglobin and methemoglobin, Arch. Biochem. Biophys. 238: 314–321.CrossRefGoogle Scholar
  24. Kelman, D.J., DeGray, J.A., and Mason, R., 1994, Reaction of myoglobin with hydrogen peroxide forms a peroxyl radical which oxidizes substrates, J. Biol. Chem. 269: 7458–7463.Google Scholar
  25. King, N.K., and Winfield, M.E., 1963, The mechanism of metmyoglobin oxidation, J. Biol. Chem. 238: 1520–1528.Google Scholar
  26. Larahjinha, J., Almeida, L., and Madeira, V., 1995, Reduction of ferrylmyoglobin by dietary phenolic acid derivatives of cinamic acid, Free Radic. Biol. Med. 19: 329–337.CrossRefGoogle Scholar
  27. Larahjinha, J., Vieira, O., Almeida, L., and Madeira, V., 1996, Inhibition of metmyoglobin/H2O2-dependent low density lipoprotein lipid peroxidation by naturally occurring phenolic acids, Biochem. Pharmacol. 51: 395–402.CrossRefGoogle Scholar
  28. Mondai, M.S., and Mitra, S., 1996, Kinetic studies of the two-step reactions of H2O2 with manganese-reconstituted myoglobin, Biochim. Biophys. Acta, 1296: 174–180.CrossRefGoogle Scholar
  29. McCord, J., 1985, Oxygen-derived free radicals in post-ischemic tissue injury, New Engl. J. Med. 312: 159–163.CrossRefGoogle Scholar
  30. Newman, E.S.R., Rice-Evans, C.A., and Davies, M., 1991, Identification of initiating agents in myoglobin-induced lipid peroxidation, Biochem. Biophys. Res. Commun. 179: 1414–1419.CrossRefGoogle Scholar
  31. Osawa, Y., and Korzekwa, K., 1991, Oxidative modification by low levels of HOOH can transform myoglobin to an oxidase, Proc. Natl. Acad. Sci. USA 88: 7081–7085.CrossRefGoogle Scholar
  32. Osawa, Y., and Williams, M.S., 1996, Covalent crosslinking of the heme prosthetic group to myoglobin by H2O2: Toxicological implications, Free Radic. Biol. Med. 21: 35–41.CrossRefGoogle Scholar
  33. Ostdal, H., Daneshvar, B., and Skibsted, L.H., 1996, Reduction of ferrylmyoglobin by b-lactoglobulin, Free Radic. Res. 24: 429–438.CrossRefGoogle Scholar
  34. Puppo, A., and Halliwell, B., 1988, Formation of hydroxyl radicals in biological systems. Does myoglobin stimulate hydroxyl radical formation from hydrogen peroxide? Free Radic. Res. Commun. 6: 415–422.CrossRefGoogle Scholar
  35. Rao, S.I., Wilks, A., and Ortiz de Montellano, P.R., 1993, The role of His-64, Tyr-103, Tyr-146, and Tyr-151 in the epoxidation of styrene and beta-methylstyrene by recombinant sperm whale myoglobin, J. Biol. Chem. 268: 803–809.Google Scholar
  36. Rao, S.I., Wilks, A., Hamberg, M., and Ortiz de Montellano, P.R., 1994, The lipoxygenase activity of myoglobin. Oxidation of linoleic acid by the ferryl oxygen rather than protein radical, J. Biol. Chem. 269: 7210–7216.Google Scholar
  37. Romero, F.J., Ordonez, I., Arduini, A., and Cadenas, E., 1992, The reactivity of thiols and disulfides with different redox states of myoglobin, J. Biol. Chem. 267: 1680–1688.Google Scholar
  38. Sundarsan, M., Yu, Z-X., Ferrans, V.J., Irani, K., and Finkel, T., 1995, Requirement for generation of HO2 for platelet-derived growth factor signal transduction, Science, 270: 296–299.CrossRefGoogle Scholar
  39. Turner, J.J.O., Rice-Evans, C.A., Davies, M., and Newman, E.S.R., 1991, The formation of free radicals by cardiac myocytes under oxidative stress and the effects of electron-donating drugs, Biochem. J. 277: 833–837.Google Scholar
  40. Wilks A., and Ortiz de Montellano, P.R., 1992, Intramolecular translocation of the protein radical formed in the reaction of recombinant sperm whale myoglobin with H2O2, J. Biol. Chem. 267: 8827–8833.Google Scholar
  41. Yonetani, T., and Schleyer, H., 1967, Studies on cytochrom c peroxidase. IX. The reaction of ferrylmyoglobin with hydroperoxides and a comparison of peroxide-induced compounds of ferrylmyoglobin and cytochrom c peroxidase, J. Biol. Chem. 242: 1974–1979.Google Scholar
  42. Yusa, K., and Shikama, K., 1987, Oxidation of oxymyoglobin to metmyoglobin with hydrogen peroxide: Involvement of ferryl intermediate, Biochemistry 26: 6684–6688.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1998

Authors and Affiliations

  • Dimitrios Galaris
    • 1
  • Stelianos Kokkoris
    • 1
  • Ioannis Toumpoulis
    • 1
  • Panagiotis Korantzopoulos
    • 1
  1. 1.Laboratory of Biological ChemistryUniversity of Ioannina Medical SchoolIoanninaGreece

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