Inhibition of Escherichia coli inorganic pyrophosphatase by fructose-1-phosphate
- 76 Downloads
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.
Keywordspyrophosphatase fructose-1-phosphate regulation site-directed mutagenesis allostery
Unable to display preview. Download preview PDF.
- 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.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.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
- 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.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
- 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