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T Kulakovskaya

Publications and source records attributed to T Kulakovskaya.

3 recordsLinked to original sources

Polyphosphate and phosphate pump.

In microbial cells, inorganic polyphosphate (polyP) plays a significant role in increasing cell resistance to unfavorable environmental conditions and in regulating different biochemical processes. polyP is a polyfunctional compound. The most important of its functions are the following: phosphate and energy reservation, cation sequestration and storage, membrane channel formation, participation in phosphate transport, involvement in cell envelope formation and function, gene activity control, regulation of enzyme activities, and a vital role in stress response and stationary-phase adaptation. The functions of polyP have changed greatly during the evolution of living organisms. In prokaryotes, the most important functions are as an energy source and a phosphate reserve. In eukaryotic microorganisms, the regulatory functions predominate. Therefore, a great difference is observed between prokaryotes and eukaryotes in their polyP-metabolizing enzymes. Some key prokaryotic enzymes are not present in eukaryotes, and conversely, eukaryotes have developed new polyP-metabolizing enzymes that are not present in prokaryotes. The synthesis and degradation of polyP in each specialized organelle and compartment of eukaryotic cells are mediated by different sets of enzymes. This is consistent with the endosymbiotic hypothesis of eukaryotic cell origin.

Biological Evolution↗

Membrane-bound and soluble polyphosphatases of mitochondria of Saccharomyces cerevisiae: identification and comparative characterization.

Isolated mitochondria of Saccharomyces cerevisiae possess polyphosphatases insensitive to a number of inhibitors of ATPase and pyrophosphatase of the same organelles and differing from the last two by neutral pH optima and molecular masses. After subfractionation of mitochondria, the polyphosphatase activity is distributed among the membrane and soluble preparations. The membrane-bound and soluble polyphosphatase activities are represented by different enzymes distinguished by molecular masses, substrate specificity, Km values, and relation to mono- and divalent cations. The membrane-bound polyphosphatases have molecular masses of 120 and 76 kDa, and the soluble one of about 36 kDa. All three enzymes appear to have a monomeric structure. The soluble polyphosphatase activity is stimulated by divalent cations in contrast to the membrane-bound one which is inhibited by the same cations, including Mg2+. Monovalent cations do not actually change the activity of the soluble enzyme, but stimulate it in the membrane preparation. Specific activities for the hydrolysis of polyphosphates with average chain lengths of 9-188 phosphate residues increase under increasing degree of substrate polymerization in the membrane preparation and are actually unchanged in the soluble one. The affinity of the soluble enzyme to polyphosphates is 5-10 times higher than that of the membrane-bound polyphosphatases.

Acid Anhydride Hydrolases↗

Purification and properties of polyphosphatase from Saccharomyces cerevisiae cytosol.

A homogenous polyphosphatase preparation was obtained from Saccharomyces cerevisiae cytosol with a 3.8% yield and 3540-fold purification. The enzyme hydrolysed polyphosphate (polyP) with various chain lengths, including polyP3, and split Pi off the end of the chain. It was inactive with respect to ATP, PPi, and p-nitrophenylphosphate. Its specific activity with polyP15 was 283 U/mg protein. The polyphosphatase was a monomeric protein with a molecular mass of 40 kDa. This enzyme was inactive without divalent cations and with Cu2+ and Ca2+. The ability of other divalent cations to activate the enzyme decreased in the following order; Co2+ > Mn2+ > Mg2+ > Zn2+. A kinetic model of the hydrolysis of polyP3 and action of Mg2+ is proposed.

Acid Anhydride Hydrolases↗