The substrate specificities of sunflower and soybean phospholipases D using transphosphatidylation reaction

Lipids in Health and Disease

Research

Abdelkarim Abousalham¹  Slim Abdelkafi² 

[1] 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;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;Université de Sfax, Centre de Biotechnologie de Sfax, Laboratoire des Bio-Procédés Environnementaux, Sfax, Tunisia;

关键词: Acyl Chain;    Phosphatidic Acid;    Transphosphatidylation Reaction;    Transphosphatidylation Activity;    Phospholipid Polar Head Group;   

DOI  :  10.1186/1476-511X-10-196

 received in 2011-09-25, accepted in 2011-11-01,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundPhospholipase D (PLD) belongs to a lipolytic enzyme subclass which catalyzes the hydrolysis and transesterification of glycerophospholipids at the terminal phosphodiester bond.ResultsIn 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.ConclusionsThe substrate specificity is modulated by the fatty acid composition of the phosphatidylcholine used as well as by the presence of other charged phospholipids.

【 授权许可】

CC BY   
© Abdelkafi and Abousalham; licensee BioMed Central Ltd. 2011