Enzyme Dynamics in Nonaqueous Media at Subzero Temperatures

  • Pierre Douzou


Despite the continuous development of new data acquisition techniques that reduce the time and improve the precision of recording, it is not possible to directly observe the nature and structure of most intermediates or to determine rates of conformational changes that accompany each activation process, stepping the reaction pathway either in solution or in the crystalline state. In order to circumvent this difficulty, we set up a procedure to investigate enzyme-catalyzed reactions at subzero temperatures in which the reaction rates are dramatically slowed according to the classic Arrhenius relation. In particular, reactions that normally occurred in a few seconds or minutes could be sufficiently slowed to permit enzyme—substrate intermediates to be stabilized and eventually isolated by column chromatography, and then studied by available spectroscopic techniques, including x-ray diffraction by enzyme crystals. The physicochemical basis of the procedure as well as its preliminary results have been reviewed elsewhere (1–3).


Elementary Step Subzero Temperature Nonaqueous Medium Enzyme Dynamics Collision Complex 
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  1. 1.
    Douzou, P. (1977) Cryobiochemistry: An Introduction. Academic Press, New York.Google Scholar
  2. 2.
    Douzou, P. (1974) Methods Biochem. Anal. 22: 401–512.PubMedCrossRefGoogle Scholar
  3. 3.
    Douzou, P. (1977) Adv. Enzymol. 45: 157–272.PubMedGoogle Scholar
  4. 4.
    Douzou, P., Hui Bon Hoa, G., Maurel, F., and Travers, F. (1976) In Handbook of Biochemistry and Molecular Biology, 3rd ed. Vol. I, (edited by G. D. Fasman) pp. 520–539. CRC PressGoogle Scholar
  5. 5.
    Maurel, P., Hui Bon Hoa, G., and Douzou, P. (1975) J. Biol. Chem. 250: 1376 1382.Google Scholar
  6. 6.
    Larroque, C., Maurel, P., Balny, C., and Douzou, P. (1976) Anal. Biochem. 73: 9–19.PubMedCrossRefGoogle Scholar
  7. 7.
    Maurel, P., and Douzou, P. (1975) J. Biol. Chem. 250: 2678–2682.PubMedGoogle Scholar
  8. 8.
    Cox, R. P. (1977) Biochem. Soc. Trans. 6: 689–697.Google Scholar
  9. 9.
    Douzou, P., and Petsko, G. A. (1984) Adv. Protein Chem. 36: 245–361.PubMedCrossRefGoogle Scholar
  10. 10.
    Douzou, P., Sireix, R., and Travers, F. (1970) Proc. Natl. Acad. Sci. USA 66: 787–790.PubMedCrossRefGoogle Scholar
  11. 11.
    Barman, T. E., and Travers, F. (1985) Methods Biochem. Anal. 31: 1–59.PubMedCrossRefGoogle Scholar
  12. 12.
    Douzou, P. (1982) Proc. R. Soc. Lond. 217: 1–28.PubMedCrossRefGoogle Scholar
  13. 13.
    Fink, A. L. (1976) J. Theor. Biol. 61: 419–445.PubMedCrossRefGoogle Scholar
  14. 14.
    Douzou, P. (1979) Q. Rev. Biophysi. 14: 521–569.CrossRefGoogle Scholar
  15. 15.
    Debey, P., Balny, C., and Douzou, P. (1976) FEBS Lett. 69: 231–236.PubMedCrossRefGoogle Scholar
  16. 16.
    Fink, A. L. (1973) Biochemistry 12: 1736–1742.PubMedCrossRefGoogle Scholar
  17. 17.
    Biosca, J. R., Travers, F., Hillaire, D., and Barman, T. E. (1984) Biochemistry 151: 47–50.Google Scholar
  18. 18.
    Yagi, K., Lange, R., and Douzou, P. (1980) Biochem. Biophys. Res. Commun. 97: 370–374.PubMedCrossRefGoogle Scholar
  19. 19.
    Balny, C., Travers, F., and Douzou, P. (1987) Submitted.Google Scholar
  20. 20.
    Marden, M. C., and Hui Bon Hoa, G. (1982) Eur. J. Biochem. 129: 111–1117.PubMedCrossRefGoogle Scholar
  21. 21.
    Dreyfus, M., Vandenbunder, B., and Buc. H. (1978) FEBS Lett. 95: 185–189.PubMedCrossRefGoogle Scholar
  22. 22.
    Brody, E. N., and Leautey, J. (1973) Eur. J. Biochem. 36: 347–361.PubMedCrossRefGoogle Scholar
  23. 23.
    Crepin, M., Cukier-Kahn, R., and Gros, F. (1973) Proc. Natl. Acad. Sci. USA 72: 333–337.CrossRefGoogle Scholar
  24. 24.
    Hamel, E., and Nakamoto, T. (1977) Biochemistry 11: 223–238.Google Scholar
  25. 25.
    Nakanishi, S., Adhya, S., Gottesman, M., Pastan, L. (1974) J. Biol. Chem. 249: 4050–4056.PubMedGoogle Scholar
  26. 26.
    Ballesta, J. P., and Vasquez, D. (1973) Biochemistry 12: 5063–5068.PubMedCrossRefGoogle Scholar
  27. 27.
    Voigt, J., Sander, G., Nagel, K., and Parmeggiani, A. (1974) Biochem. Biophys. Res. Commun. 57: 1279–1286.PubMedCrossRefGoogle Scholar
  28. 28.
    Hui Bon Hoa, G., Begard, E., Beaudry, P., Maurel, P., Grunberg-Manago, M., and Douzou, P. (1980) Biochemistry 19: 3080–3088.CrossRefGoogle Scholar

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© Springer-Verlag New York Inc. 1988

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  • Pierre Douzou

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