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Plant Physiology 85:1151-1155 (1987)
© 1987 American Society of Plant Biologists

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Development and Growth Regulation

Phosphatidylinositol Specific Phospholipase C of Plant Stems 1

Membrane Associated Activity Concentrated in Plasma Membranes

Helmut Pfaffmann, Elmar Hartmann, Andrew O. Brightman and D. James Morré

Institute of General Botany, University of Mainz, D-6500 Mainz, Federal Republic of Germany, Department of Medicinal Chemistry and Pharmacognosy, Purdue University, West Lafayette, Indiana 47907

A phosphatidylinositol-specific phospholipase C of plant stems (EC 3.1.4.10) assayed at pH 6.6 and at 30°C cleaved phosphatidylinositol such that more than 85% of the product was inositol-1-phosphate. Other phospholipids were cleaved 5 to 10% or less under these conditions. The phospholipase had both a soluble and a membrane-associated form. The soluble activity accounted for approximately 85 to 90% of the activity and 15% was associated with membranes. The membrane-associated activity was most concentrated in the plasma membranes of hypocotyl segments of both soybean (Glycine max) and bushbean (Phaseolus vulgaris). The plasma membrane location was verified by analysis of highly purified plasma membranes prepared both by aqueous two-phase partitioning and by preparative free-flow electrophoresis and from the quantitation of the activity in all major cell fractions. Internal membranes also contained phospholipase C activity but at specific activity levels of about 0.1 those present in plasma membranes. Golgi apparatus-enriched fractions from which plasma membrane contaminants were removed by two-phase partition contained the activity at specific activity levels 0.2 those of plasma membrane. Both the soluble and the membrane-associated activity was stimulated by calcium but not by calmodulin, either alone or in the presence of calcium.


1 Supported in part by a grant 8206222 from the National Science Foundation (to D.J.M.).







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Copyright © 1987 by the American Society of Plant Biologists