Partial purification and characterization of membrane-associated diacylglycerol kinase of Drosophila heads

H Inoue; T Yoshioka; Y Hotta

doi:10.1016/0167-4838(92)90327-a

Partial purification and characterization of membrane-associated diacylglycerol kinase of Drosophila heads

Biochim Biophys Acta. 1992 Jul 31;1122(2):219-24. doi: 10.1016/0167-4838(92)90327-a.

Authors

H Inoue¹, T Yoshioka, Y Hotta

Affiliation

¹ Department of Basic Sciences, School of Human Sciences, Waseda University, Tokorozawa, Japan.

PMID: 1322704
DOI: 10.1016/0167-4838(92)90327-a

Abstract

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-polyacrylamide 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 microM for ATP and 540 microM for diolein, respectively. The activity of the DG kinase was inhibited by deoxycholate and was not activated by Ca2+.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Membrane / enzymology
Deoxycholic Acid / pharmacology
Diacylglycerol Kinase
Drosophila / enzymology*
Hydrogen-Ion Concentration
Kinetics
Phosphotransferases / antagonists & inhibitors
Phosphotransferases / chemistry
Phosphotransferases / isolation & purification*

Substances

Deoxycholic Acid
Phosphotransferases
Diacylglycerol Kinase