Abstract
The kinetic theory of the substrate reaction during modification of enzyme activity previously described by Tsou [Tsou (1988),Adv. Enzymol. Relat. Areas Mol. Biol. 61, 381–436] has been applied to a study of the kinetics of the course of inactivation of the mitochondrial succinate-ubiquinone reductase by 5,5′-dithiobis-(2-nitro-benzoic acid) (DTNB). The results show that the inactivation of this enzyme by DTNB is a conformation-change-type inhibition which involves a conformational change of the enzyme before inactivation. The microscopic rate constants were determined for the reaction of the inactivator with the enzyme. The presence of the substrate provides marked protection of this enzyme against inactivation by DTNB. The modification reaction of the enzyme using DTNB was shown to follow a triphasic course by following the absorption at 412 nm. Among these reactive thiol groups, the fast-reaction thiol group is essential for the enzyme activity. The results suggest that the essential thiol group is situated at the succinate-binding site of the mitochondrial succinate-ubiquinone reductase.
Similar content being viewed by others
References
Ackrell, B. A. C., Kearney, E. B., Coles, C. J., Singer, T. P., Beinert, H., Wan, Y.-P., and Folkers, K. (1977).Arch. Biochem. Biophys. 182, 107–117.
Ackrell, B. A. C., Ball, M. B., and Kearney, E. B. (1980).J. Biol. Chem. 255, 2761–2769.
Baginsky, M. L., and Hatefi, Y. (1969).J. Biol. Chem. 244, 5313–5319.
Beinert, H. Ackrell, B. A. C., Kearney, E. B., and Singer, T. P. (1975).Eur. J. Biochem. 54, 185–194.
Davis, K. A., and Hatefi, Y. (1971).Biochemistry 10, 2509–2516.
Ellman, G. L. (1959).Arch. Biochem. Biophys. 82, 70–77.
Gornall, A. D., Bardawill, C. J., and David, M. M. (1949).J. Biol. Chem. 177, 751–756.
Hatefi, Y., and Galante, Y. M. (1980).J. Biol. Chem. 255, 5530–5537.
Hatefi, Y., and Stiggall, D. L. (1978).Meth. Enzymol. 53, 27–35.
King, T. E. (1967).Meth. Enzymol. 10(58), 322–331.
Liu, C. and Tsou, C. L. (1992).Biochem. J. 282, 501–504.
Ohnishi, T. (1987).Curr. Topics Bioenerg. 15, 37–64.
Ohnishi, T., Lim, J., Winter, D. B., and King, T. E. (1976).J. Biol. Chem. 251, 2105–2109.
Tsou, C. L. (1988).Adv. Enzymol. Relat. Areas Mol. Biol. 61, 381–436.
Tushurashvili, P. R., Garcibova, E. V., Ledenev, A. N., and Vinogridov, A. D. (1985).Biochim. Biophys. Acta 809, 145–159.
Vinogradov, A. D., and King, T. E. (1979).Meth. Enzymol. 15, 118–127.
Vinogradov, A. D., Ackrell, B. A. C., and Singer, T. P. (1975).Biochem. Biophys. Res. Commun. 67, 803–809.
Wang, Z. X., Preiss, B., and Tsou, C. L. (1988).Biochemistry 27, 5095–5100.
Yu, L., and Yu, C. A. (1982).J. Biol. Chem. 257, 2016–2021.
Ziegler, D. M., and Doeg, K. A. (1962).Arch. Biochim. Biophys. 97, 41–50.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Yang, Y., Wang, HR., Xu, JX. et al. Kinetics of modification of the mitochondrial succinate-ubiquinone reductase by 5,5′-dithiobis-(2-nitro-benzoic acid). J Protein Chem 15, 169–176 (1996). https://doi.org/10.1007/BF01887397
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01887397