Phospholipase-induced modulation of dolichyl-phosphomannose synthase activity.
Rat liver dolichyl-phosphomannose synthase is optimally active when the enzyme is reconstituted with lipids that prefer a nonlamellar macroscopic organization in isolation, such as phosphatidylethanolamine (PE), but the enzyme is only negligibly active in the presence of lipids that normally form stable bilayers, such as phosphatidylcholine (PC) [Jensen, J.W., & Schutzbach, J.S. (1985) Eur. J. Biochem. 153, 41-48]. We now report that the activity of the synthase can be modulated by incorporating diacylglycerol and lysophosphatidylcholine into the lipid matrix. Enzyme activity in PC bilayers was stimulated by the presence of diacylglycerol, a lipid that has a conical dynamic molecular shape and disrupts bilayer stability. In PC/diacylglycerol mixtures the apparent Km for dolichyl-P was 30-fold lower than the apparent Km for the polyprenol acceptor in PC membranes. Enzyme activity was also stimulated when diacylglycerol was generated in situ by incubation of PC vesicles with phospholipase C. In contrast, the activity of enzyme reconstituted in PE dispersions, or in PE/PC bilayers, was markedly inhibited by the presence of lysophospholipids. Enzyme activity was also reduced by the in situ generation of lysophospholipids in PE/PC vesicles by incubation with phospholipase A2. Since lysophospholipids and diacylglycerols arise in vivo as products of phospholipid metabolism, modulation of enzyme activity by these compounds may represent a potential regulatory mechanism for the synthesis of oligosaccharide lipids.
Jensen
J W
JW
Department of Microbiology, University of Alabama, Birmingham 35294.
Schutzbach
J S
JS
D000818:Animals; D006602:Hexosyltransferases / Q000378:metabolism*; D007700:Kinetics; D008081:Liposomes; D008244:Lysophosphatidylcholines; D008364:Mannosyltransferases / Q000378:metabolism*; D008862:Microsomes, Liver / Q000201:enzymology*; D010743:Phospholipids / Q000494:pharmacology*; D051381:Rats; D013329:Structure-Activity Relationship; D010738:Type C Phospholipases / Q000378:metabolism*
D008081:Liposomes; D008244:Lysophosphatidylcholines; D010743:Phospholipids; D006602:Hexosyltransferases; D008364:Mannosyltransferases; C022755:dolichyl-phosphate beta-D-mannosyltransferase; D010738:Type C Phospholipases