The purification of nucleoside diphosphate kinase from rat brain.
Nucleoside diphosphate kinase from the cytosol of rat brain was purified to electrophoretic homogeneity through a series of chromatography columns, including DE-52, AcA-34 Ultrogel, phenyl-Sepharose and Q-Sepharose. The molecular weight of the enzyme was 64,000 as estimated by gel filtration, and it consisted of 3 identical 19,000 subunits. [35S]-GTP gamma S binding and autophosphorylation of the enzyme, and inhibition of the GTPase activity of G-protein were used to screen for the enzyme. These methods were shown to be useful for purifying and assaying the enzyme.
Zhang
D
D
Institute of Basic Medical Sciences, CAMS, Beijing.
Chang
K J
KJ
D000818:Animals; D001921:Brain / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology; D020558:GTP Phosphohydrolases / Q000378:metabolism; D008970:Molecular Weight; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry / Q000302:isolation & purification*; D010766:Phosphorylation; D051381:Rats
D009701:Nucleoside-Diphosphate Kinase; D020558:GTP Phosphohydrolases