Abstract
An iron-hexacyanide-covered microelectrode sensor has been used to continuously monitor the kinetics of hydrogen peroxide decomposition catalyzed by oxidized cytochrome oxidase. At cytochrome oxidase concentration ≈1 μM, the catalase activity behaves as a first order process with respect to peroxide at concentrations up to ≈300–400 μM and is fully blocked by heat inactivation of the enzyme. The catalase (or, rather, pseudocatalase) activity of bovine cytochrome oxi- dase is characterized by a second order rate constant of ≈2•102 M-1•sec-1 at pH 7.0 and room temperature, which, when divided by the number of H2O2 molecules disappearing in one catalytic turnover (between 2 and 3), agrees reasonably well with the second order rate constant for H2O2-dependent conversion of the oxidase intermediate FI-607 to FII-580. Accordingly, the catalase activity of bovine oxidase may be explained by H2O2 procession in the oxygen-reducing center of the enzyme yielding superoxide radicals. Much higher specific rates of H2O2 decomposition are observed with preparations of the bacterial cytochrome c oxidase from Rhodobacter sphaeroides. The observed second order rate constants (up to ≈3000 M-1•sec-1) exceed the rate constant of peroxide binding with the oxygen-reducing center of the oxidized enzyme (≈500 M-1•sec-1) several-fold and therefore cannot be explained by catalytic reaction in the a 3/CuB site of the enzyme. It is proposed that in the bacterial oxidase, H2O2 can be decomposed by reacting with the adventitious transition metal ions bound by the polyhistidine-tag present in the enzyme, or by virtue of reaction with the tightly-bound Mn2+, which in the bacterial enzyme substitutes for Mg2+ present in the mitochondrial oxidase.
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Tsukihara, T., Aoyama, H., Yamashita, E., Takashi, T., Yamaguichi, H., Shinzawa-Itoh, K., Nakashima, R., Yaono, R., and Yoshikawa, S. (1996) Science, 272, 1136–1144.
Ostermeier, C., Harrenga, A., Ermler, U., and Michel, H. (1997) Proc. Natl. Acad. Sci. USA, 94, 10547–10553.
Svensson-Ek, M., Abramson, J., Larsson, G., Tornroth, S., Brzezinski, P., and Iwata, S. (2002) J. Mol. Biol., 321, 329–339.
Qin, L., Hiser, C., Mulichak, A., Garavito, R. M., and Ferguson-Miller, S. (2006) Proc. Natl. Acad. Sci. USA, 103, 16117–16122.
Soulimane, T., Buse, G., Bourenkov, G. B., Bartunik, H. D., Huber, R., and Than, M. E. (2000) EMBO J., 19, 1766–1776.
Ferguson-Miller, S., and Babcock, G. T. (1996) Chem. Rev., 7, 2889–2907.
Belevich, I., and Verkhovsky, M. I. (2008) Antioxidants and Redox Signaling, 10, 1–29.
Einarsdottir, O., and Szundi, I. (2004) Biochim. Biophys. Acta, 1655, 263–273.
Konstantinov, A. A., Siletsky, S., Mitchell, D., Kaulen, A., and Gennis, R. B. (1997) Proc. Natl. Acad. Sci. USA, 94, 9085–9090.
Vygodina, T. V., Pecoraro, C., Mitchell, D., Gennis, R., and Konstantinov, A. A. (1998) Biochemistry, 37, 3053–3061.
Bickar, D., Bonaventura, J., and Bonaventura, C. (1982) Biochemistry, 21, 2661–2666.
Wrigglesworth, J. (1984) Biochem. J., 217, 715–719.
Vygodina, T. V., and Konstantinov, A. A. (1987) FEBS Lett., 219, 387–392.
Vygodina, T., and Konstantinov, A. (1989) Biochim. Biophys. Acta, 973, 390–398.
Vygodina, T. V., and Konstantinov, A. A. (1988) Ann. NY Acad. Sci., 550, 124–138.
Vygodina, T. V., and Konstantinov, A. A. (2007) Biochemistry (Moscow) 72, 1056–1064.
Pecoraro, C., Gennis, R. B., Vygodina, T. V., and Konstantinov, A. A. (2001) Biochemistry, 40, 9695–9708.
Proshlyakov, D. A., Pressler, M. A., DeMaso, C., Leykam, J. F., DeWitt, D. L., and Babcock, G. L. (2000) Science, 290, 1588–1591.
Ksenzenko, M. Y., Berka, V., Vygodina, T. V., Ruuge, E. K., and Konstantinov, A. A. (1992) FEBS Lett., 297, 63–66.
Konstantinov, A. A., Capitanio, N., Vygodina, T. V., and Papa, S. (1992) FEBS Lett., 312, 71–74.
Kato, S., Ueno, T., Fukuzumi, S., and Watanabe, Y. (2004) J. Biol. Chem., 279, 52376–52381.
Marquez, L. A., Huang, J. T., and Dunford, H. B. (1994) Biochemistry, 33, 1447–1454.
Kettle, A. J., and Winterbourn, C. C. (2001) Biochemistry, 40, 10204–10212.
Orii, Y., and Okunuki, K. (1963) J. Biochem., 54, 207–213.
Gorren, A. C. F., Dekker, H., and Wever, R. (1985) Biochim. Biophys. Acta, 809, 90–96.
Bergmayer, H. U., Gawehn, K., and Grassl, M. (1970) in Methoden der Enzymatischen Analyze, Vol. 1 (Bergmayer, H. U., ed.) Verlag Chemie, Weinheim.
Sellers, R. M. (1980) Analyst, 105, 950–954.
Nourooz-Zadeh, J. (1999) Meth. Enzymol., 300, 58–62.
Puganova, E. A., Komarov, A. V., Vagin, M. Yu., Karyakina, E. E., and Karyakin, A. A. (2004) Nano- and Microsystem Technique (Moscow), 12, 42–44.
Karyakin, A. A., Gitelmacher, O. V., and Karyakina, E. E. (1994) Analyt. Lett., 27, 2861–2869.
Karyakin, A. A. (2007) in Electrochemical Sensors, Biosensors and Their Biomedical Applications (Zhang, X., Ju, H., and Wang, J., eds.) Academic Press (Elsevier), Amsterdam-New York, p. 616.
Karyakin, A. A., and Karyakina, E. E. (1999) Sensors and Actuators, B. 57, 268–273.
Fowler, L. R., Richardson, S. H., and Hatefi, Y. (1962) Biochim. Biophys. Acta, 64, 170–173.
Mitchell, D. M., and Gennis, R. B. (1995) FEBS Lett., 368, 148–150.
Chakrabortya, S., and Raj, C. R. (2009) Biosensors and Bioelectronics, 24, 3264–3268.
Weng, L., and Baker, G. M. (1991) Biochemistry, 30, 5727–5733.
Qin, L., Liu, J., Mills, D. A., Proshlyakov, D. A., Hiser, C., and Ferguson-Miller, S. (2009) Biochemistry, 48, 5121–5130.
Sharpe, M. A., Krzyaniak, M. D., Xu, S., McCracken, J., and Ferguson-Miller, S. (2009) Biochemistry, 48, 328–335.
Barynin, V. V., Whittaker, M. M., Antonyuk, S., Lamzin, V. S., Harrison, P. M., Artymiuk, P. J., and Whittaker, J. W. (2001) Structure, 9, 725–738.
McClune, G. J., and Fee, J. A. (1976) FEBS Lett., 67, 294–298.
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Published in Russian in Biokhimiya, 2010, Vol. 75, No. 11, pp. 1533–1543.
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Bolshakov, I.A., Vygodina, T.V., Gennis, R. et al. Catalase Activity of Cytochrome c Oxidase Assayed with Hydrogen Peroxide-Sensitive Electrode Microsensor. Biochemistry Moscow 75, 1352–1360 (2010). https://doi.org/10.1134/S0006297910110064
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DOI: https://doi.org/10.1134/S0006297910110064