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+.
| 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 | |
| Publication status | Published - 1992 Jul 31 |
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Keywords
- (Drosophila)
- Diacylglycerol kinase
- Enzyme characterization
- Enzyme purification
- Membrane-associated enzyme
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
- Structural Biology
Cite this
Partial purification and characterization of membrane-associated diacylglycerol kinase of Drosophila heads. / Inoue, Hiroko; Yoshioka, Tohru; Hotta, Yoshiki.
In: Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular, Vol. 1122, No. 2, 31.07.1992, p. 219-224.Research output: Contribution to journal › Article
}
TY - JOUR
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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UR - http://www.scopus.com/inward/citedby.url?scp=0026664532&partnerID=8YFLogxK
U2 - 10.1016/0167-4838(92)90327-A
DO - 10.1016/0167-4838(92)90327-A
M3 - Article
C2 - 1322704
AN - SCOPUS:0026664532
VL - 1122
SP - 219
EP - 224
JO - Biochimica et Biophysica Acta - Proteins and Proteomics
JF - Biochimica et Biophysica Acta - Proteins and Proteomics
SN - 1570-9639
IS - 2
ER -