[Purification and characteristics of Saccharomyces cerevisiae cytosol polyphosphatase].
The polyphosphatase with specific activity of 283 units/mg was purified 3450-fold to homogeneity with 3.8% yield from cytosol of Saccharomyces cerevisiae yeast. Polyphosphatase is monomeric 40 kD protein. The enzyme hydrolyzes polyphosphates of various chain length including tripolyphosphate but ATP, pyrophosphate, and p-nitrophenyl phosphate are not the substrates. Enzyme activity is maximal at 50 degrees C and pH 6.5-8.5. Several cations of bivalent metals stimulated the enzyme activity 8-66-fold (Co2+ > Mn2+ > Mg2+ > Zn2+ > Fe2+). The enzyme is inactive in the presence of Ca2+ or Cu2+. Heparin, antibodies against cell-envelope polyphosphatase, and Cu2+ or Zn2+ in the presence of Mg2+ are potent inhibitors of cytosolic polyphosphatase. Cytosolic polyphosphatase is similar to purified cell-envelope polyphosphatase but differs in some properties from nuclear, vacuolar, and mitochondrial polyphosphatase of the very same yeast.
Andreeva
N A
NA
Kulakovskaia
T V
TV
Kulaev
I S
IS
D017766:Acid Anhydride Hydrolases / Q000302:isolation & purification* / Q000378:metabolism; D002412:Cations; D002852:Chromatography, Ion Exchange; D003600:Cytosol / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature
D002412:Cations; D007202:Indicators and Reagents; D017766:Acid Anhydride Hydrolases; C022563:endopolyphosphatase