Skip to main content

Coupling of proteolysis to ATP hydrolysis uponEscherichia coli lon protease functioning: I. kinetic aspects of ATP hydrolysis

Russian Journal of Bioorganic Chemistry volume 26, pages 474–481 (2000)Cite this article

Abstract

Some aspects of theEscherichia coli Lon protease ATPase function were studied around the optimum pH value. It was revealed that in the absence of the protein substrate the maximum ATPase activity of the enzyme is observed at an equimolar ratio of ATP and Mg2+ ions in the area of their millimolar concentrations. Free components of the substrate complex (ATP-Mg)2− inhibit the enzyme ATPase activity. It is hypothesized that the effector activity of free Mg2+ ions is caused by the formation of the “ADP-Mg-form” of ATPase centers. It was shown that the activation of ATP hydrolysis in the presence of the protein substrate is accompanied by an increase in the affinity of the (ATP-Mg)2− complex to the enzyme, by an elimination of the inhibiting action of free Mg2+ ions without altering the efficiency of catalysis of ATP hydrolysis (based on thek cat value), and by a change in the type of inhibition of ATP hydrolysis by the (ADP-Mg)− complex (without changing theK i value). Interaction of the Lon protease protein substrate with the enzyme area located outside the peptide hydrolase center was demonstrated by a direct experiment.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

References

  1. Gottesman, S. and Maurizi, M.R.,Microbiol. Rev., 1992, vol. 56, pp. 592–621.

    PubMed  CAS  Google Scholar 

  2. Gottesman, S.,Ann. Rev. Genet., 1996, vol. 30, pp. 465–506.

    PubMed  Article  CAS  Google Scholar 

  3. Goldberg, A.L.,Eur. J. Biochem., 1992, vol. 203, pp. 9–23.

    PubMed  Article  CAS  Google Scholar 

  4. Grossman, A.D., Erickson, J.W., and Gross, C.A.,Cell, 1984, vol. 38, pp. 383–390.

    PubMed  Article  CAS  Google Scholar 

  5. Goff, S.A., Casson, L.P., and Goldberg, A.L.,Proc. Natl. Acad. Sci. USA, 1984, vol. 81, pp. 6647–6651.

    PubMed  Article  CAS  Google Scholar 

  6. Phillips, T.A., van Bogelen, R.A., and Neidhardt, F.C.,J. Bacteriol., 1984, vol. 159, pp. 283–287.

    PubMed  CAS  Google Scholar 

  7. Baker, T.A., Grossman, A.D., and Gross, C.A.,Proc. Natl. Acad. Sci. USA, 1984, vol. 81, pp. 6779–6783.

    PubMed  Article  CAS  Google Scholar 

  8. Misuzawa, S. and Gottesman, S.,Proc. Natl. Acad. Sci. USA, 1983, vol. 80, pp. 358–362.

    Article  Google Scholar 

  9. Stout, V., Torres-Cabassa, A.S., Maurizi, M.R., Gutnick, D., and Gottesman, S.,J. Bacteriol., 1991, vol. 173, pp. 1738–1747.

    PubMed  CAS  Google Scholar 

  10. Dierksen, K.P., Marks, J., Chen, D.D., and Trempy, J.E.,J. Bacteriol., 1994, vol. 176, pp. 5126–5130.

    PubMed  CAS  Google Scholar 

  11. Van Melderen, L., Thi, M.H.D., Lecchi, P., Gottesman, S., Couturier, M., and Maurizi, M.R.,J. Biol. Chem., 1996, vol. 271, pp. 27730–27738.

    PubMed  Article  Google Scholar 

  12. Rotanova, T.V.,Bioorg. Khim., 1999, vol. 25, pp. 883–891.

    PubMed  CAS  Google Scholar 

  13. Amerik, A.Yu., Antonov, V.K., Ostroumova, N.I., Rotanova, T.V., and Chistyakova, L.G.,Bioorg. Khim., 1990, vol. 16, pp. 869–881.

    PubMed  CAS  Google Scholar 

  14. Charette, M.F., Henderson, G.W., Kezdy, F.J., and Markovitz, A.,J. Mol. Biol., 1982, vol. 162, pp. 503–510.

    PubMed  Article  CAS  Google Scholar 

  15. Amerik, A.Yu., Antonov, V.K., Gorbalenya, A.E., Kotova, S.A., Rotanova, T.V., and Shimbarevich, E.V.,FEBS Lett., 1991, vol. 287, pp. 211–214.

    PubMed  Article  CAS  Google Scholar 

  16. Goldberg, A.L. and Waxman, L.,J. Biol. Chem., 1985, vol. 260, pp. 12029–12034.

    PubMed  CAS  Google Scholar 

  17. Edmunds, T. and Goldberg, A.L.,J. Cell Biochem., 1986, vol. 32, pp. 186–191.

    Article  Google Scholar 

  18. Menon, A.S. and Goldberg, A.L.,J. Biol. Chem., 1987, vol. 262, pp. 14921–14928.

    PubMed  CAS  Google Scholar 

  19. Waxman, L. and Goldberg, A.L.,Proc. Natl. Acad. Sci. USA, 1982, vol. 79, pp. 4883–4887.

    PubMed  Article  CAS  Google Scholar 

  20. Menon, A.S., Waxman, L., and Goldberg, A.L.,J. Biol. Chem., 1987, vol. 262, pp. 722–726.

    PubMed  CAS  Google Scholar 

  21. Gottesman, S., Maurizi, M.R., and Wickner, S.,Cell, 1997, vol. 91, pp. 435–438.

    PubMed  Article  CAS  Google Scholar 

  22. Menon, A.S. and Goldberg, A.L.,J. Biol. Chem., 1987, vol. 262, pp. 14929–14934.

    PubMed  CAS  Google Scholar 

  23. Waxman, L. and Goldberg, A.L.,Science, 1986, vol. 232, pp. 500–503.

    PubMed  Article  CAS  Google Scholar 

  24. Maurizi, M.R.,Experientia, 1992, vol. 48, pp. 178–200.

    PubMed  Article  CAS  Google Scholar 

  25. Bencini, D.A., Wild, J.R., and O’Donovan, G.A.,Anal. Biochem., 1983, vol. 132, pp. 254–258.

    PubMed  Article  CAS  Google Scholar 

  26. Fisher, H. and Glockshuber, R.,J. Biol. Chem., 1993, vol. 268, pp. 22502–22507.

    Google Scholar 

  27. Lemasters, J.J. and Hackenbrock, Ch.R.,Biochemistry, 1977, vol. 16, pp. 445–447.

    PubMed  Article  CAS  Google Scholar 

  28. Smith, C.K., Baker, T.A., and Sauer, R.T.,Proc. Natl. Acad. Sci. USA, 1999, vol. 96, pp. 6678–6682.

    PubMed  Article  CAS  Google Scholar 

  29. Leffers, G., Rasulova, F., Kessel, M., Steven, A.C., Leapman, R., Gottesman, S., and Maurizi, M.R., Abstracts of Papers,Meet. on Biology of Proteolysis. Cold Spring Harbor, NY, May 5–9, 1999, p. 52.

  30. Goldberg, A.L., Moerschell, R.P., Chung, C.H., and Maurizi, M.R.,Methods Enzymol., 1994, vol. 244, pp. 350–375.

    PubMed  CAS  Article  Google Scholar 

  31. Laemmli, U.K.,Nature, 1970, vol. 227, pp. 680–685.

    PubMed  Article  CAS  Google Scholar 

  32. Bradford, M.M.,Anal. Biochem., 1976, vol. 72, pp. 248–254.

    PubMed  Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117871, GSP-7 Moscow, Russia

    E. E. Melnikov, K. B. Tsirulnikov & T. V. Rotanova

Authors
  1. E. E. Melnikov
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. K. B. Tsirulnikov
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. T. V. Rotanova
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T. V. Rotanova.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Melnikov, E.E., Tsirulnikov, K.B. & Rotanova, T.V. Coupling of proteolysis to ATP hydrolysis uponEscherichia coli lon protease functioning: I. kinetic aspects of ATP hydrolysis. Russ J Bioorg Chem 26, 474–481 (2000). https://doi.org/10.1007/BF02758618

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02758618

Key words

  • ATPase
  • ATP-dependent proteolysis
  • lon gene
  • E. coli
  • Lon protease