A phospholipid requirement for dolichol pyrophosphate N-acetylglucosamine synthesis in phospholipase A2-treated rat lung microsomes.
The role of phospholipids in the activity of UDP-Glc-NAc:dolichol phosphate GlcNAc-1-phosphate transferase of rat lung microsomes has been investigated. Treatment of microsomes with phospholipase A2 in the presence of delipidated bovine serum albumin resulted in a time-dependent loss of 65 to 75% of the enzyme activity and approximately 30% of the phospholipids. Addition of phosphatidylglycerol to the enzyme assay system containing phospholipase A2-treated microsomes restored activity to that obtained with native microsomes and phosphatidylglycerol. Addition of phosphatidylinositol, phosphatidylcholine, or cardiolipin resulted in only partial restoration of activity, whereas phosphatidylserine and phosphatidylethanolamine were without effect. Triton X-100 was not by itself capable of restoring activity, but was required for the phospholipid effect. Measurements of the phospholipase A2 hydrolysis products released from the microsomes during digestion, and other control experiments of adding fatty acids and lysophospholipids to the enzyme assay system, indicated that the loss of UDP-GlcNAc:dolichol phosphate GlcNAc-1-phosphate transferase activity was not due to product inhibition.
D000818:Animals; D002308:Cardiolipins / Q000494:pharmacology; D008168:Lung / Q000201:enzymology*; D008297:Male; D008861:Microsomes / Q000201:enzymology; D010714:Phosphatidylethanolamines / Q000494:pharmacology; D010718:Phosphatidylserines / Q000494:pharmacology; D010740:Phospholipases / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D010743:Phospholipids / Q000494:pharmacology*; D010770:Phosphotransferases / Q000378:metabolism*; D011103:Polyisoprenyl Phosphate Monosaccharides / Q000096:biosynthesis*; D011105:Polyisoprenyl Phosphate Sugars / Q000096:biosynthesis*; D051381:Rats; D011919:Rats, Inbred Strains; D013997:Time Factors; D017855:Transferases (Other Substituted Phosphate Groups)
D002308:Cardiolipins; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D010743:Phospholipids; D011103:Polyisoprenyl Phosphate Monosaccharides; D011105:Polyisoprenyl Phosphate Sugars; C018439:N-acetylglucosaminylpyrophosphoryldolichol; D010770:Phosphotransferases; D017855:Transferases (Other Substituted Phosphate Groups); C023222:UDPacetylglucosamine-dolichyl-phosphate acetylglucosamine-1-phosphate transferase; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2