title: Purification and Characterization of Membrane-Bound Inositol Phospholipid-Specific Phospholipase C from Suspension-Cultured Rice (Oryza sativa L.) Cells (Identification of a Regulatory Factor).
abstract: A membrane-bound inositol phospholipid-specific phospholipase C was solubilized from rice (Oryza sativa L.) microsomal membranes and purified to apparent homogeneity using a series of chromatographic separations. The apparent molecular mass of the enzyme was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 42,000 D, and the isoelectric point was 5.1. The optimum pH for the enzyme activity was approximately 6.5, and the enzyme was activated by both Ca2+ and Sr2+. The chemical and catalytic properties of the purified membrane-bound phospholipase C differed from those of the soluble enzyme reported previously (K. Yotsushima, K. Nakamura, T. Mitsui, I. Igaue [1992] Biosci Biotech Biochem 56: 1247-1251). In addition, we found a regulatory factor for the phosphatidylinositol-4,5-bisphosphate (PIP2) hydrolyzing activity of phospholipase C from rice cells. The regulatory factor was dissociated from the catalytic subunit of phospholipase C during the purification. The regulatory factor was necessary to induce PIP2-hydrolyzing activity of both membrane-bound and -soluble phospholipase C; these purified enzymes had no activity alone. Because the plasma membranes isolated from rice cells could also act as a regulatory factor, the regulatory factor seems to be localized in the plasma membranes. Regulation of inositol phospholipid turnover in rice cells is discussed.
authors.affiliation: Department of Biosystem Science, Graduate School of Science and Technology (K.Y.), and Department of Applied Biological Chemistry, Faculty of Agriculture (T.M., T.T., T.H., I.I.), Niigata University, 2-Ikarashi, Niigata 950-21, Japan.
authors.forename: K.
authors.initials: K
authors.lastname: Yotsushima
authors.forename: T.
authors.initials: T
authors.lastname: Mitsui
authors.forename: T.
authors.initials: T
authors.lastname: Takaoka
authors.forename: T.
authors.initials: T
authors.lastname: Hayakawa
authors.forename: I.
authors.initials: I
authors.lastname: Igaue