A membrane-associated diacylglycerol kinase of Drosophila heads was purified to near homogeneity from the KCl extract of Drosophila heads. The purification procedure involved chromatography on Q-Sepharose, ammonium sulfate fractionation, Superose 12, hydroxyapatite and ATP-agarose. Sodium dodecyl sulfate-polyacrylamode gel electrophoresis of fractions after the ATP-agarose column chromatography showed that only a 115 kDa protein correlated well with the enzyme activity. The apparent Km values of partially purified DG kinase were 220 μM for ATP and 540 μM for diolein, respectively. The activity of the DG kinase was inhibited by deoxycholate and was not activated by Ca2+.
|Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
|Published - 1992 7月 31
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