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PARTIAL PURIFICATION OF DROSOPHILA GLUTAMATE DECARBOXYLASE 1
Author(s) -
Chude Obi,
Roberts Eugene,
Wu JangYen
Publication year - 1979
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1979.tb11078.x
Subject(s) - chromatography , sephadex , chemistry , hydroxylapatite , enzyme , potassium phosphate , dithiothreitol , homogenization (climate) , specific activity , phosphate , phosphate buffered saline , biochemistry , biology , biodiversity , ecology
— A 600‐fold purification of l ‐glutamatc decarboxylasc from Drosophila melanogaster Oregon R Wild Type has been achieved. The purification procedures involve the initial homogenization of whole flies in dilute potassium phosphate buffer containing dithiothreitol and phenylmethyl sulfonyl fluoride as protectors, followed by a series of column chromatography with hydroxylapatite, Sephadex G‐150 and DEAE‐Sephadex. The purified enzyme has an apparent K m of 11 m m for l ‐glutamate and requires a 40m m ‐K + for maximum activity. The purified enzyme shows only 1 pH optimum around pH 7.5, while crude preparations of the inset display 2 pH optima, pH 4.8–5.2. and 7.5. The significance and possible application of this study are also discussed.