title: [A functionally important Tyr-89 residue in Saccharomyces cerevisiae pyrophosphatase. II. A possible role in the mechanism of enzyme action].
abstract: Earlier it has been demonstrated that inactivation of inorganic pyrophosphatase (PPase) of S. cerevisiae by 7-chloro-4-nitronbenzofurasane is due to modification of Tyr89. The effect of pH and active center ligands on this reaction has been studied. It was found that pK for Tyr89 does not exceed 8.5; the phosphate-metal complex binding to the high affinity center protects Tyr89 from inactivation. Activating ions (Mg2+ and Zn2+) do not influence the inactivation, whereas the PPase inhibitor, Ca2+, enhances this process after saturation of the high affinity binding site. Saturation of two binding sites with Ca2+ has a protective effect on the enzyme. An increase in the rate of Tyr89 binding to the inhibitor in the presence of low concentrations of Ca2+ is due to the decrease of Tyr89 pK. The data obtained suggest that Tyr89 is located near the high affinity binding site for phosphate. The high reactivity of Tyr89 and its tight binding in the active center point to the presence of a hydrogen bondage with the substrate and suggest a role of a proton donor whose acceptor is the product of the enzymatic reaction, i.e., phosphate.
authors.forename: A V
authors.initials: AV
authors.lastname: Raznikov
authors.forename: N A
authors.initials: NA
authors.lastname: Steriopolo
authors.forename: V A
authors.initials: VA
authors.lastname: Skliankina
authors.forename: S M
authors.initials: SM
authors.lastname: Avaeva
mesh_terms: D001572:Benzofurans / Q000494:pharmacology; D001665:Binding Sites; D002118:Calcium / Q000737:chemistry; D002413:Cations, Divalent; D043564:Inorganic Pyrophosphatase; D010710:Phosphates / Q000737:chemistry; D011755:Pyrophosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D001572:Benzofurans; D002413:Cations, Divalent; D010710:Phosphates; C043708:7-chloro-4-nitrobenzofuran; D014443:Tyrosine; D011755:Pyrophosphatases; D043564:Inorganic Pyrophosphatase; D002118:Calcium
vernacular_title: Funktsional'no vazhnyĭ ostatok Tyr-89 pirofosfatazy S. cerevisiae. II. Vozmozhnaia rol' v mekhanizme deĭstviia fermenta.