Purification and Characterization of Phosphatidylinositol-specific Phospholipase C in Suspension-cultured Cells of Rice (Oryza sativa L.)
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概要
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Phosphatidylinositol-specific phospholipase C was purified from the soluble fraction of suspension-cultured rice cells. The apparent molecular weight of rice enzyme was estimated to be 50,000 by both Sephadex G-100 gel filtration and SDS-polyacrylamide gel electrophoresis, indicating that the enzyme is composed of a single polypeptide. The enzyme had an isoelectric point of 6.3. The soluble phospholipase C had a high degree of specificity toward phosphatidylinositol and a weak activity toward phosphatidyl-inositol monophosphate, while the enzyme did not hydrolyze the other phospholipids or p-nitrophenylphosphorylcholine. V_<max> and K_m values were 5.0 μmol/min/mg protein and 0.3 mM, respectively. The pH dependency of the enzyme activity was sharp with an optimum of 5.2. In addition, the phospholipase C was a Ca^<2+>-dependent enzyme. The marked activation of enzyme was observed in the presence of 10 to 250μM Ca^<2+> and higher Ca^<2+> concentrations than 1 mM had a strong inhibitory effect. A possible regulation of the phospholipase C activity by pH and Ca^<2+> concentrations in the rice cells is discussed.
- 社団法人日本農芸化学会の論文
- 1992-08-23
著者
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NAKAMURA Kouji
Department of Astronomical Science, The Graduate University for Advanced Studies
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Mitsui Toshiaki
Department Of Applied Biological Chemistry Faculty Of Agriculture Niigata University
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Mitsui Toshiaki
Department Of Agricultural Chemistry Faculty Of Agriculture Niigata University
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Igaue Ikuo
Department Of Agricultural Chemistry Faculty Of Agriculture Niigata University
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Igaue Ikuo
Department Of Applied Biological Chemistry Faculty Of Agriculture Niigata University
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Yotsushima Kenji
Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata University
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Yotsushima Kenji
Department Of Biosystem Science Graduate School Of Science And Technology
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Nakamura Kouji
Department Of Astronomical Science The Graduate University For Advanced Studies
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Nakamura Kouji
Department Of Applied Biological Chemistry Faculty Of Agriculture Niigata University
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