Purification and Characterization of Phosphatidylinositol-specific Phospholipase C in Suspension-cultured Cells of Rice (<i>Oryza sativa</i>L.)

Yotsushima, Kenji; Nakamura, Kouji; Mitsui, Toshiaki; Igaue, Ikuo

doi:10.1271/bbb.56.1247

Cited by 17 publications

(14 citation statements)

References 13 publications

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“…This suggests that the membrane-bound PLC is a key enzyme of the signal transduction system in higher plants. Previously, we reported the characterization of the soluble inositol phospholipid-specific PLC enzyme purified from suspension-cultured rice (Oryza sativa L.) cells (Yotsushima et al, 1992). In the present study, we have purified and characterized a membrane-bound PLC from rice cells and found a regulatory factor for the PIPz-hydrolyzing activity of PLC.…”

mentioning

confidence: 62%

“…PI, ~-a-[myoinositol-2-~H(N)] (37-185 GBq/mmol), PIP[myoinosit01-2-~H(N)] (37-185 GBq/mmol), and PIPz [myoinositol-2-3H(N)] (37-1 85 GBq/mmol) were obtained from American Radiolabeled Chemicals (St. Louis, MO). The suspensioncultured cells of rice (Oryza sativa L. cv Nipponkai) were prepared as described previously (Yotsushima et al, 1992 Plant Physiol. Vol.…”

Section: Methodsmentioning

confidence: 99%

“…Vol. 102, 1993 Enzyme and Protein Assays A standard assay of PLC was essentially identical with the procedure described before (Yotsushima et al, 1992). The reaction mixture for the soluble enzyme consisted of 50 mM acetate buffer (pH 5.2), 0.1 mM CaC12, 0.08% (w/v) sodium deoxycholate, 1 mM PI, and enzyme in a final volume of 75 pL.…”

Section: Methodsmentioning

confidence: 99%

“…This suggests that the inositol phospholipid turnover is involved in signal transduction of higher plants. Two different types of inositol phospholipid-specific PLC, soluble and membrane-bound enzymes, have been identified in severa1 plant cells (McMurry and Irvine, 1987;Melin et al, 1987;Pfaffmann et al, 1987;Tate et al, 1989;Kamada and Muto, 1991;Yotsushima et al, 1992). The membrane-bound PLC preferentially hydrolyzes PIPl and PIP over PI, whereas the soluble PLC is not able to hydrolyze PIP2 (McMurry and Irvine, 1987;Melin et al, 1987;Yotsushima et al, 1992).…”

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confidence: 99%

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Purification and Characterization of Membrane-Bound Inositol Phospholipid-Specific Phospholipase C from Suspension-Cultured Rice (Oryza sativa L.) Cells (Identification of a Regulatory Factor)

et al. 1993

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mentioning

confidence: 62%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Purification and Characterization of Membrane-Bound Inositol Phospholipid-Specific Phospholipase C from Suspension-Cultured Rice (Oryza sativa L.) Cells (Identification of a Regulatory Factor)

et al. 1993

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“…As seen in Fig. 4, NPC4 showed very low activity for PIP 2 (Ͻ10% of PC-hydrolyzing activity), suggesting that NPC4 has different substrate selectivity from PI-PLC (32,33). Next, we analyzed the effect of divalent cations on the enzyme activity, since many plant phospholipases are characterized as Ca 2ϩ -dependent enzymes.…”

Section: Npc4 Encoded a Functional Pc-plc That Prefers Pc For The Submentioning

confidence: 99%

A Novel Phosphatidylcholine-hydrolyzing Phospholipase C Induced by Phosphate Starvation in Arabidopsis

Nakamura

Awai

Masuda

et al. 2005

Journal of Biological Chemistry

231

294

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During phosphate starvation, it is known that phospholipids are degraded, and conversely, a nonphosphorus galactolipid digalactosyldiacylglycerol accumulates in the root plasma membrane of plants. We report a novel phospholipase C that hydrolyzes phosphatidylcholine and is greatly induced in response to phosphate deprivation in Arabidopsis. Since phosphatidylcholinehydrolyzing activity by phospholipase C was highly upregulated in phosphate-deprived plants, gene expression of some phospholipase C was expected to be induced during phosphate starvation. Based on amino acid sequence similarity to a bacterial phosphatidylcholine-hydrolyzing phospholipase C, six putative phospholipase Cs were identified in the Arabidopsis genome, one of which, NPC4, showed significant transcriptional activation upon phosphate limitation. Molecular cloning and functional expression of NPC4 confirmed that the NPC4 gene encoded a functional phosphatidylcholinehydrolyzing phospholipase C that did not require Ca 2؉ for its activity. Subcellular localization analysis showed that NPC4 protein was highly enriched in the plasma membrane. Analyses of transferred DNA-tagged npc4 mutants revealed that disruption of NPC4 severely reduces the phosphatidylcholine-hydrolyzing phospholipase C activity in response to phosphate starvation. These results suggest that NPC4 plays an important role in the supply of both inorganic phosphate and diacylglycerol from membrane-localized phospholipids that would be used for phosphate supplementation and the replacement of polar lipids in the root plasma membrane during phosphate deprivation.Phosphorus is an essential element for plant growth, development, and reproduction. It plays decisive roles not only in regulation of various enzymes but also in constitutive components such as membrane phospholipids and nucleic acids. In most soils, despite its abundance, phosphorus is not freely available for assimilation by roots (1). Therefore, plants have developed distinct systems to cope with phosphate deficiency.When plants suffer from phosphate limitation, highly integrated systems are activated both for assimilation of P i and supplementation of P i from innermost phosphorus storage. The former action is represented by a morphological modification of root architecture upon P i starvation, which presumably facilitates P i uptake by enlargement of absorptive root surface areas (2). On the other hand, the latter has been described by the dynamic evolution of metabolism that is altered toward the supply of free P i . During P i starvation, overall phospholipid content that corresponds to 30% of total P i storage is decreased, and conversely, contents of nonphosphorus galactolipid, digalactosyldiacylglycerol (DGDG), 1 increase significantly (3). Galactolipids such as monogalactosyldiacylglycerol (MGDG) and DGDG are ubiquitous in plants, but are typically found only in plastids, especially in photosynthetic membranes (4). These galactolipids are synthesized by the galactosylation of diacylglycerol (DAG) by MGDG synthase an...

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Determination of Phospholipase C Activity In Vitro

Hernández‐Sotomayor

Muñoz-Sánchez

2013

Methods in Molecular Biology

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Measurement of phospholipase C (PLC) activity in vitro is a valuable biochemistry technique easily applicable in samples from different organisms. It quantifies the enzymatic activity of a key protein involved in critical developmental functions in organisms such as plants, animals, and bacteria. A protocol is described which assays the formation of two main products of the PLC hydrolysis reaction on radioactively labeled phospholipid substrates.

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Purification and Characterization of Phosphatidylinositol-specific Phospholipase C in Suspension-cultured Cells of Rice (Oryza sativaL.)

Cited by 17 publications

References 13 publications

Purification and Characterization of Membrane-Bound Inositol Phospholipid-Specific Phospholipase C from Suspension-Cultured Rice (Oryza sativa L.) Cells (Identification of a Regulatory Factor)

Purification and Characterization of Membrane-Bound Inositol Phospholipid-Specific Phospholipase C from Suspension-Cultured Rice (Oryza sativa L.) Cells (Identification of a Regulatory Factor)

A Novel Phosphatidylcholine-hydrolyzing Phospholipase C Induced by Phosphate Starvation in Arabidopsis

Determination of Phospholipase C Activity In Vitro

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