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Inhibition of Inorganic Pyrophosphatase from Escherichia coli with Inorganic Phosphate

Abstract

The interaction of inorganic pyrophosphatase from E. coli with inorganic phosphate (P i) was studied in a wide concentration range of phosphate. The apoenzyme gives two inactive compounds with P i, a product of phosphorylation of the carboxylic group of the active site and a stable complex, which can be detected in the presence of the substrate. The phosphorylation occurs when P i is added on a millimole concentration scale, and micromole concentrations are sufficient for the formation of the complex. The formation of the phosphorylated enzyme was confirmed by its sensitivity to hydroxylamine and a change in the properties of the inactive enzyme upon its incubation in alkaline medium. The phosphorylation of pyrophosphatase and the formation of the inactive complex occur upon interaction of inorganic phosphate with different subsites of the enzyme active sites, which are connected by cooperative interactions.

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Grigorieva, O.V., Mit'kevich, V.A., Sklyankina, V.A. et al. Inhibition of Inorganic Pyrophosphatase from Escherichia coli with Inorganic Phosphate. Russian Journal of Bioorganic Chemistry 27, 27–33 (2001). https://doi.org/10.1023/A:1009574901422

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  • DOI: https://doi.org/10.1023/A:1009574901422

  • inorganic pyrophosphatase
  • phosphorylation
  • inactivation
  • cooperativity