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Purification and properties of exopolyphosphatase isolated from Saccharomyces cerevisiae vacuoles
Author(s) -
Andreeva Nadezhda A,
Kulakovskaya Tatiana V,
Kulaev Igor S
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00591-2
Subject(s) - saccharomyces cerevisiae , biochemistry , pyrophosphate , cytosol , divalent , vacuole , molecular mass , enzyme , yeast , activator (genetics) , inorganic pyrophosphatase , gtp' , chemistry , pyrophosphatase , biology , organic chemistry , gene , cytoplasm
An exopolyphosphatase (polyPase) with a specific activity of 60 U/mg protein has been purified from the vacuolar sap of Saccharomyces cerevisiae . The molecular mass of the intact enzyme was found to be 245 kDa. It is highly specific towards high‐molecular polyphosphates (polyP). The activity with polyP 9 is 24% of that with polyP 208 . The apparent K m for polyP 15 and polyP 208 hydrolysis is 93 and 2.4 μM, respectively. The enzyme is slightly active with polyP 3 and adenosine‐5′‐tetraphosphate, but does not hydrolyze pyrophosphate, ATP, GTP and p ‐nitrophenylphosphate. It is stimulated by divalent metal cations. Co 2+ , the best activator, stimulates it 6‐fold. Antibodies that inhibit the cell envelope and cytosol polyPases of S. cerevisiae have no effect on the vacuolar polyPase. The vacuolar polyPase differs from other yeast polyPases in molecular mass, substrate specificity and effects of activators.