[Purification and properties of a catalytically active fragment of soluble nucleoside triphosphatase from bovine kidney]
| Autor: | Makarchikov Af |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
GTP'
Cations Divalent Size-exclusion chromatography Uridine Triphosphate Kidney Biochemistry Catalysis Substrate Specificity Column chromatography Animals Inosine Triphosphate chemistry.chemical_classification Nucleoside-triphosphatase Chromatography Extraction (chemistry) Hydrogen-Ion Concentration Ribonucleotides Nucleoside-Triphosphatase Peptide Fragments Kinetics Protein Subunits Enzyme chemistry Solubility Biocatalysis Chromatography Gel Specific activity Cattle Electrophoresis Polyacrylamide Gel Guanosine Triphosphate |
| Zdroj: | Ukrains'kyi biokhimichnyi zhurnal (1999 ). 85(3) |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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