PMID:11706190

Citation	Wang, C and Wang, X (2001) A novel phospholipase D of Arabidopsis that is activated by oleic acid and associated with the plasma membrane. Plant Physiol. 127:1102-12
Abstract	Oleate-dependent phospholipase D (PLD; EC 3.1.4.4) has been reported in animal systems, but its molecular nature is unkown. Multiple PLDs have been characterized in plants, but none of the previously cloned PLDs exhibits the oleate-activated activity. Here, we describe the biochemical and molecular identification and characterization of an oleate-activated PLD in Arabidopsis. This PLD, designated PLDdelta, was associated tightly with the plasma membrane, and its level of expression was higher in old leaves, stems, flowers, and roots than in young leaves and siliques. A cDNA encoding the oleate-activated PLD was identified, and catalytically active PLDdelta was expressed from its cDNA in Escherichia coli. PLDdelta was activated by free oleic acid in a dose-dependent manner, with the optimal concentration being 0.5 mM. Other unsaturated fatty acids, linoleic and linolenic acids, were less effective than oleic acid, whereas the saturated fatty acids, stearic and palmitic acids, were totally ineffective. Phosphatidylinositol 4,5-bisphosphate stimulated PLDdelta to a lesser extent than oleate. Mutation at arginine (Arg)-611 led to a differential loss of the phosphatidylinositol 4,5-bisphosphate-stimulated activity of PLDdelta, indicating that separate sites mediate the oleate regulation of PLDdelta. Oleate stimulated PLDdelta's binding to phosphatidylcholine. Mutation at Arg-399 resulted in a decrease in oleate binding by PLDdelta and a loss of PLDdelta activity. However, this mutation bound similar levels of phosphatidylcholine as wild type, suggesting that Arg-399 is not required for PC binding. These results provide the molecular information on oleate-activated PLD and also suggest a mechanism for the oleate stimulation of this enzyme.
Links	PubMed PMC129279
Keywords	Amino Acid Sequence; Arabidopsis/enzymology; Arabidopsis/genetics; Calcium/metabolism; Cell Membrane/metabolism; Cloning, Molecular; Enzyme Activation; Expressed Sequence Tags; Gene Expression Regulation, Plant; Molecular Sequence Data; Mutagenesis; Oleic Acid/metabolism; Phosphatidylcholines/metabolism; Phospholipase D/chemistry; Phospholipase D/classification; Phospholipase D/metabolism; Phylogeny; Sequence Alignment
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
ARATH:PLDD1		GO:0004630: phospholipase D activity	ECO:0000315:		F		Figure 5- Shows phospholipase activity levels in WT and mutated gene	complete CACAO 4258
ARATH:PLDD1	enables	GO:0004630: phospholipase D activity	ECO:0000315: mutant phenotype evidence used in manual assertion		F		Seeded From UniProt	complete

References

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