title: The substrate specificities of sunflower and soybean phospholipases D using transphosphatidylation reaction.
abstract: BACKGROUND
  Phospholipase D (PLD) belongs to a lipolytic enzyme subclass which catalyzes the hydrolysis and transesterification of glycerophospholipids at the terminal phosphodiester bond.
  
  
  RESULTS
  In this work, we have studied the substrate specificity of PLDs from germinating sunflower seeds and cultured-soybean cells, using their capacity of transphosphatidylation. In the presence of a nucleophilic acceptor, such as [¹⁴C]ethanol, PLD catalyzes the production of phosphatidyl-[¹⁴C]-ethanol. The resulting product is easily identified since it is well separated from the other lipids by thin-layer chromatography. The main advantage of this assay is that the phospholipid used as substrate does not need to be radiolabelled and thus allow us a large choice of polar heads and fatty acids. In vitro, we observed that sunflower and soybean cell PLD show the following decreasing order of specificity: phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol; while phosphatidylserine and phosphatidylinositol are utilized much less efficiently.
  
  
  CONCLUSIONS
  The substrate specificity is modulated by the fatty acid composition of the phosphatidylcholine used as well as by the presence of other charged phospholipids.
authors: Organization and Dynamics of Biological Membranes, UMR 5246 ICBMS, CNRS-Université Claude Bernard Lyon 1, Bâtiment Raulin, 43, boulevard du 11 novembre 1918, 69622 Villeurbanne, Cedex, France.
authors: Slim
authors: S
authors: Abdelkafi
authors: Abdelkarim
authors: A
authors: Abousalham
mesh_terms: D018929:Cell Culture Techniques; D002852:Chromatography, Ion Exchange; D057075:Enzyme Assays; D020404:Glycerophospholipids / Q000737:chemistry*; D006368:Helianthus / Q000201:enzymology*; D006868:Hydrolysis; D007700:Kinetics; D010739:Phospholipase D / Q000737:chemistry* / Q000302:isolation & purification; D036226:Seedlings / Q000201:enzymology*; D013025:Glycine max / Q000166:cytology / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D020404:Glycerophospholipids; D010739:Phospholipase D