title: Specificity of lysophospholipase D.
abstract: The specificity of lysophospholipase D (1-alkyl-sn-glycero-3-phosphoethanolamine ethanolaminehydrolase, EC 3.1.4.39; also works on choline analogs) for 1-alkyl- and 1-acyl-linked substrates was examined using rat liver microsomes. The microsomes were treated with diisopropylphosphorofluoridate to inhibit the hydrolysis of acyl chains from the acyl-linked compounds (1-palmitoyl-sn-glycero-3-phosphocholine and 1-palmitoyl-sn-glycero-3-phosphoethanolamine) and were treated with p-bromophenacyl bromide to block acylation of the compounds tested. In the presence of the inhibitors, 1-alkyl-sn-glycero-3-phosphocholine and 1-alkyl-sn-glycero-3-phosphoethanolamine were hydrolyzed extensively by lysophospholipase D but the corresponding 1-acyl-linked analogs were only negligibly hydrolyzed. Lysophospholipase D therefore appears to be specific for the ether-linked compounds. 1-Alk-1-'-enyl-sn-glycero-3-phosphoethanolamine (lyso plasmalogen) was also tested as a substrate, but a plasmalogenase in the rat liver microsomes rapidly hydrolyzed the compound and we were unable to determine whether it is a substrate for lysophospholipase D. Alkyl-linked substrates containing long-chain acyl groups at the 2-position are not hydrolyzed by the enzymes. We tested 1-alkyl-2-acetoyl-sn-glycero-3-phosphocholine and 1-alkyl-2-acetoyl-sn-glycero-3-phosphoethanolamine to determine if the less bulky, more hydrophilic acetate group would permit hydrolysis by lysophospholipase D; the derivatives did not appear to be attacked, except after hydrolysis of the acetate group. However, in the absence of inhibitors, the acetate groups were rapidly hydrolyzed by microsomal preparations.
mesh_terms: D000818:Animals; D004791:Enzyme Inhibitors; D008245:Lysophospholipase / Q000378:metabolism*; D008862:Microsomes, Liver / Q000201:enzymology* / Q000378:metabolism; D010740:Phospholipases / Q000378:metabolism*; D010955:Plasmalogens / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity
chemical_list: D004791:Enzyme Inhibitors; D010955:Plasmalogens; D010740:Phospholipases; D008245:Lysophospholipase