The phosphorylcholine acceptor in the phosphatidylcholine:ceramide cholinephosphotransferase reaction. Is the enzyme a transferase or a hydrolase?
The cholinephosphotransferase reaction is shown to be catalyzed by an enzyme which has no hydrolytic activity and which is different from a phospholipase C type activity also present in these plasma membrane preparations. Diacylglycerols and sphingosine, at a concentration above 0.4 mM, are effective inhibitors of sphingomyelin formation in the presence of 0.3 mM free ceramide, the true acceptor in this reaction. Free sphingosine is not an acceptor for the cholinephosphate group, as the anticipated reaction product, sphingosylphosphocholine , could not be detected. Sphingosine inhibition may result from its structural similarity to the natural substrates of the reaction, ceramide and diacylglycerols. From the data obtained with cholesterol, triacylglycerols, acetylated ( triacetyl ) sphingosine and acetylated ceramides used as potential inhibitors of the reaction it is concluded that the free hydroxyl group at C1 of the sphingosine backbone or of the glycerol moiety of diacylglycerols and a non-polar residue consisting of an aliphatic chain were prerequisites for inhibitory activity. These results are discussed in terms of substrate specificity of the enzyme catalyzing the transfer reaction. Some of the factors influencing the regulation of the phosphatidylcholine/sphingomyelin ratio in the plasma membrane were related to the topography of sphingomyelin in the outer half-layer of the plasma membrane.
D000818:Animals; D002518:Ceramides / Q000378:metabolism; D002798:Diacylglycerol Cholinephosphotransferase / Q000378:metabolism*; D004075:Diglycerides / Q000378:metabolism; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008345:Manganese / Q000378:metabolism; D051379:Mice; D010770:Phosphotransferases / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013109:Sphingomyelins / Q000096:biosynthesis; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000378:metabolism
D002518:Ceramides; D004075:Diglycerides; D013109:Sphingomyelins; D012710:Serum Albumin, Bovine; D008345:Manganese; D010770:Phosphotransferases; D002798:Diacylglycerol Cholinephosphotransferase; D010738:Type C Phospholipases