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

^*この研究の対応する著者

研究成果: Article › 査読

5 被引用数 (Scopus)

抄録

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⁺.

本文言語	English
ページ（範囲）	219-224
ページ数	6
ジャーナル	Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
巻	1122
号	2
DOI	https://doi.org/10.1016/0167-4838(92)90327-A
出版ステータス	Published - 1992 7月 31

ASJC Scopus subject areas

生化学
生物理学
分子生物学
構造生物学

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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+.",

keywords = "(Drosophila), Diacylglycerol kinase, Enzyme characterization, Enzyme purification, Membrane-associated enzyme",

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

year = "1992",

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doi = "10.1016/0167-4838(92)90327-A",

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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

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