Biochemistry (Moscow)

, Volume 82, Issue 8, pp 953–956 | Cite as

Inhibition of Escherichia coli inorganic pyrophosphatase by fructose-1-phosphate

  • N. N. VorobyevaEmail author
  • S. A. Kurilova
  • V. A. Anashkin
  • E. V. Rodina


Pyrophosphate regulates vital cellular reactions, and its level in E. coli cells is under the ultimate control of inorganic pyrophosphatase. The mechanisms involved in the regulation of pyrophosphatase activity still need to be elucidated. The present study demonstrated that fructose-1-phosphate inhibits pyrophosphatase activity by a mechanism not involving competition with substrate for binding to the active site. The inhibition constant governing the binding of the inhibitor to the enzyme–substrate complex is 1.1 mM. Substitutions of Lys112, Lys115, Lys148, and Arg43 in the regulatory site completely or partially abolished the inhibition. Thus, Fru-1-P is a physiological inhibitor of pyrophosphatase that acts via a regulatory site in this enzyme.


pyrophosphatase fructose-1-phosphate regulation site-directed mutagenesis allostery 


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  1. 1.
    Chen, J., Brevet, A., Romant, M., Leveque, F., Schmitter, J.-M., Blanquet, S., and Plateau, P. (1990) Pyrophosphatase is essential for growth of Escherichia coli, J. Bacteriol., 172, 5686–5689.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Serrano-Bueno, G., Hernandez, A., Lopez-Lluch, Perez-Castineira, J. R., Navas, P., and Serrano, A. (2013) Inorganic pyrophosphatase defects lead to cell cycle arrest and autophagic cell death through NAD+ depletion in fermenting yeast, J. Biol. Chem., 288, 13082–13092.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Heikinheimo, P., Tuominen, V., Ahonen, A.-K., Teplyakov, A., Cooperman, B. S., Baykov, A. A., Lahti, R., and Goldman, A. (2001) Toward a quantum-mechanical description of metal-assisted phosphoryl transfer in pyrophosphatase, Proc. Natl. Acad. Sci. USA, 98, 3121–3126.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Samygyna, V. R., Popov, A. N., Rodina, E. V., Vorobyeva, N. N., Lamzin, V. S., Polyakov, K. M., Kurilova, S. A., Nazarova, T. I., and Avaeva, S. M. (2001) The structures of Escherichia coli inorganic pyrophosphatase complexed with Ca2+ or CaPPi at atomic resolution and their mechanistic implications, J. Mol. Biol., 314, 633–645.CrossRefGoogle Scholar
  5. 5.
    Sitnik, T. S., Vainonen, J. P., Rodina, E. V., Nazarova, T. I., Kurilova, S. A., Vorobyeva, N. N., and Avaeva, S. M. (2003) Effectory site in Escherichia coli inorganic pyrophosphatase is revealed upon mutation at the intertrimeric interface, IUBMB Life, 55, 37–41.CrossRefPubMedGoogle Scholar
  6. 6.
    Vainonen, J. P., Vorobyeva, N. N., Rodina, E. V., Nazarova, T. I., Kurilova, S. A., Skoblov, Yu. S., and Avayeva, S. M. (2005) Metal-free PPi activates hydrolysis of MgPPi by an Escherichia coli inorganic pyrophosphatase, Biochemistry (Moscow), 70, 85–96.CrossRefGoogle Scholar
  7. 7.
    Rodina, E. V., Vorobyeva, N. N., Kurilova, S. A., Belenikin, M. S., Fedorova, N. V., and Nazarova, T. I. (2007) ATP as an effector of inorganic pyrophosphatase of Escherichia coli, Biochemistry (Moscow), 72, 110–117.Google Scholar
  8. 8.
    Rodina, E. V., Vorobyeva, N. N., Kurilova, S. A., Sitnik, T. S., and Nazarova, T. I. (2007) ATP as an effector of inorganic pyrophosphatase. The role of Lys112 residue of Escherichia coli PPase in effector binding, Biochemistry (Moscow), 72, 118–127.Google Scholar
  9. 9.
    Baykov, A. A., Volk, S. E., and Unguryte, A. (1989) Inhibition of inorganic pyrophosphatase of animal mitochondria by calcium, Arch. Biochem. Biophys., 273, 287–291.CrossRefPubMedGoogle Scholar
  10. 10.
    Maeshima, M. (1991) H+-translocating inorganic pyrophosphatase of plant vacuoles. Inhibition by Ca2+, stabilization by Mg2+ and immunological comparison with other inorganic pyrophosphatases, Eur. J. Biochem., 196, 11–17.CrossRefPubMedGoogle Scholar
  11. 11.
    Avaeva, S. M., Vorobyeva, N. N., Kurilova, S. A., Nazarova, T. I., Polyakov, K. M., Rodina, E. V., and Samygina, V. R. (2000) Mechanism of inhibition of Escherichia coli inorganic pyrophosphatase by calcium ions, Biochemistry (Moscow), 65, 373–387.Google Scholar
  12. 12.
    Oganessyan, V. Yn., Kurilova, S. A., Vorobyeva, N. N., Nazarova, T. I., Popov, A. N., Lebedev, A. A., Avaeva, S. M., and Harutyunyan, E. H. (1994) X-ray crystallographic studies of recombinant pyrophosphatase from Escherichia coli, FEBS Lett., 348, 301–304.CrossRefPubMedGoogle Scholar
  13. 13.
    Avaeva, S., Ignatov, P., Kurilova, S., Nazarova, T., Rodina, E., Vorobyeva, N., Oganessyan, V., and Harutyunyan, E. (1996) Escherichia coli inorganic pyrophosphatase: sitedirected mutagenesis of the metal binding sites, FEBS Lett., 399, 99–102.CrossRefPubMedGoogle Scholar
  14. 14.
    Josse, J. (1966) Constitutive inorganic pyrophosphatase of Escherichia coli. 1. Purification and catalytic properties, J. Biol. Chem., 241, 1938–1947.PubMedGoogle Scholar
  15. 15.
    Baykov, A. A., and Avaeva, S. M. (1981) A simple and sensitive apparatus for continuous monitoring of orthophosphate in the presence of acid-labile compounds, Anal. Biochem., 116, 1–4.CrossRefPubMedGoogle Scholar
  16. 16.
    Baykov, A. A., Bakuleva, N. P., and Rea, P. A. (1993) Steady-state kinetics of substrate hydrolysis by vacuolar H+-pyrophosphatase. A simple three-state model, Eur. J. Biochem., 217, 755–762.CrossRefPubMedGoogle Scholar
  17. 17.
    Shimada, T., Yamamoto, K., and Ishihama, A. (2011) Novel members of the Cra regulon involved in carbon metabolism in Escherichia coli, J. Bacteriol., 193, 649–659.CrossRefPubMedGoogle Scholar
  18. 18.
    Kornberg, H., and Lourenco, C. (2006) A route for fructose utilization by Escherichia coli involving the fucose regulon, Proc. Natl. Acad. Sci. USA, 103, 19496–19499.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Rodina, E., Vorobieva, N., Kurilova, S., Mikulovich, J., Vainonen, J., Aro, E. M., and Nazarova, T. (2011) Identification of new protein complexes of Escherichia coli inorganic pyrophosphatase using pull-down assay, Biochimie, 93, 1576–1583.CrossRefPubMedGoogle Scholar
  20. 20.
    Kukko, E., and Heinonen, J. (1982) The intracellular concentration of pyrophosphate in the batch culture of Escherichia coli, Eur. J. Biochem., 127, 347–349.CrossRefPubMedGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2017

Authors and Affiliations

  • N. N. Vorobyeva
    • 1
    • 2
    Email author
  • S. A. Kurilova
    • 2
  • V. A. Anashkin
    • 2
  • E. V. Rodina
    • 1
  1. 1.Faculty of ChemistryLomonosov Moscow State UniversityMoscowRussia
  2. 2.Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia

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