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

Hiroko Inoue; Tohru Yoshioka; Yoshiki Hotta

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

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

Hiroko Inoue^*, Tohru Yoshioka, Yoshiki Hotta

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

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-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 K_m 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⁺.

Original language	English
Pages (from-to)	219-224
Number of pages	6
Journal	Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume	1122
Issue number	2
DOIs	https://doi.org/10.1016/0167-4838(92)90327-A
Publication status	Published - 1992 Jul 31

Keywords

(Drosophila)
Diacylglycerol kinase
Enzyme characterization
Enzyme purification
Membrane-associated enzyme

ASJC Scopus subject areas

Biochemistry
Biophysics
Molecular Biology
Structural Biology

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10.1016/0167-4838(92)90327-A

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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-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+.",

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author = "Hiroko Inoue and Tohru Yoshioka and Yoshiki Hotta",

year = "1992",

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T1 - Partial purification and characterization of membrane-associated diacylglycerol kinase of Drosophila heads

AU - Inoue, Hiroko

AU - Yoshioka, Tohru

AU - Hotta, Yoshiki

PY - 1992/7/31

Y1 - 1992/7/31

N2 - 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+.

AB - 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+.

KW - (Drosophila)

KW - Diacylglycerol kinase

KW - Enzyme characterization

KW - Enzyme purification

KW - Membrane-associated enzyme

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

Abstract

Keywords

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