Substrate-induced membrane association of phosphatidylserine synthase from Escherichia coli.
To better establish the intracellular location of the phosphatidylserine synthase of Escherichia coli and hence better understand how it is regulated in the cell, we compared the size, function, and binding properties of the enzyme made in vitro with the enzyme found in cell lysates and with the purified enzyme. The enzyme made either in vivo or in an active form in vitro was found primarily associated with the ribosomal fraction of the cell and had the same apparent molecular mass as the purified enzyme. These results were unaffected by the presence of protease inhibitors. Addition of unsupplemented E. coli membranes or membranes supplemented with phosphatidylethanolamine did not affect the subcellular distribution of the enzyme in these experiments. However, addition of membranes supplemented with either the lipid substrate, CDP-diacylglycerol, or the lipid product, phosphatidylserine, resulted in membrane association by the enzyme rather than ribosomal association. Addition of membranes supplemented with acidic lipids also brought about membrane association, but this association was primarily ionic since it was disrupted by high salt concentrations. These results strongly suggest that the ribosomal location of this enzyme is not the result of some modification event occurring after cell lysis and that the normal functioning of the enzyme involves membrane association which is primarily induced by the presence of a membrane-associated substrate.
D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D003567:Cytidine Diphosphate Diglycerides / Q000032:analysis / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D008970:Molecular Weight; D010718:Phosphatidylserines / Q000032:analysis / Q000378:metabolism; D010770:Phosphotransferases / Q000378:metabolism*; D012270:Ribosomes / Q000201:enzymology
D003567:Cytidine Diphosphate Diglycerides; D010718:Phosphatidylserines; D010770:Phosphotransferases; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase