1. [Purification and properties of a catalytically active fragment of soluble nucleoside triphosphatase from bovine kidney].
- Author
-
Makarchikov AF
- Subjects
- Animals, Biocatalysis, Cations, Divalent chemistry, Cattle, Chromatography, Gel, Electrophoresis, Polyacrylamide Gel, Guanosine Triphosphate chemistry, Hydrogen-Ion Concentration, Inosine Triphosphate chemistry, Kidney enzymology, Kinetics, Nucleoside-Triphosphatase isolation & purification, Peptide Fragments isolation & purification, Protein Subunits isolation & purification, Ribonucleotides chemistry, Solubility, Substrate Specificity, Uridine Triphosphate chemistry, Kidney chemistry, Nucleoside-Triphosphatase chemistry, Peptide Fragments chemistry, Protein Subunits chemistry
- Abstract
A catalytic fragment of soluble NTPase has been isolated from bovine kidneys.The 236-fold purification was carried out to obtain the preparation with a specific activity of 37.7 U/mg of protein. The purification scheme included the enzyme extraction followed by four column chromatography steps. The catalytic fragment was activated with divalent metal ions, had a pH optimum of 7.0, and possessed specificity for ITP, GTP, UTP and XTP. The apparent K(m) for Mg-ITP, Mg-GTP and Mg-UTP complexes were calculated from Hanes plots to be 1.70 mM, 0.93 mM and 0.48 mM, respectively. As estimated by gel filtration and SDS-PAAGE, the catalytic fragment has Mw 54.7 kDa being composed of two identical polypeptide chains. Our results suppose soluble NTPase from bovine kidney to consist of regulatory and catalytic structural units.
- Published
- 2013
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