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

Hiroko Inoue, Tohru Yoshioka, Yoshiki Hotta

    Research output: Contribution to journalArticle

    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 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 languageEnglish
    Pages (from-to)219-224
    Number of pages6
    JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
    Volume1122
    Issue number2
    DOIs
    Publication statusPublished - 1992 Jul 31

    Fingerprint

    Diacylglycerol Kinase
    Drosophila
    Purification
    Phosphotransferases
    Head
    Membranes
    Agarose Chromatography
    Column chromatography
    Deoxycholic Acid
    Ammonium Sulfate
    Enzyme activity
    Durapatite
    Fractionation
    Chromatography
    Electrophoresis
    Sodium Dodecyl Sulfate
    Sepharose
    Adenosine Triphosphate
    Gels
    Enzymes

    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 journalArticle

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