Phosphatidylethanolamine levels and regulation of phosphatidylethanolamine N-methyltransferase.
The activity of phosphatidylethanolamine (PE) N-methyltransferase in liver microsomes, measured using endogenous microsomal PE as a substrate, was elevated 2-fold in the choline-deficient state. However, methyltransferase activity assayed in the presence of a saturating concentration of phosphatidyl-N-mono-methylethanolamine or microsomal PE was unchanged by choline deficiency. Accompanying the increase in methyltransferase activity in liver homogenates and microsomes were increased PE concentrations and an increased PE to phosphatidylcholine ratio. The concentration of other phospholipids was unchanged. Immunoblot analysis of choline-deficient and choline-supplemented rat liver microsomes using a rabbit polyclonal anti-PE N-methyltransferase antibody revealed that the amount of enzyme protein was unaltered. The regulation of methyltransferase by PE levels was also investigated in cultured hepatocytes obtained from choline-deficient rat livers. Supplementation of deficient hepatocytes with 200 microM methionine resulted in a 50% reduction in cellular PE levels over a 12-h period. PE N-methyltransferase activity assayed with endogenous PE was also reduced by 50%, but phosphatidyl-N-monomethylethanolamine-dependent activity was unchanged. A 4-h supplementation with choline did not affect PE levels or methyltransferase activity. Either methionine or choline supplementation resulted in net synthesis of cellular phosphatidylcholine. Immunoblotting of membranes from methionine-supplemented hepatocytes revealed no change in enzyme protein, a further indication that enzyme mass was constitutive, and activity was regulated by the concentration of PE.
Ridgway
N D
ND
Lipoprotein Research Group, University of Alberta, Edmonton, Canada.
Yao
Z
Z
Vance
D E
DE
D000818:Animals; D002478:Cells, Cultured; D002794:Choline / Q000494:pharmacology; D002796:Choline Deficiency / Q000201:enzymology*; D004721:Endoplasmic Reticulum / Q000201:enzymology; D006706:Homeostasis; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D008715:Methionine / Q000172:deficiency* / Q000494:pharmacology; D008780:Methyltransferases / Q000378:metabolism*; D008862:Microsomes, Liver / Q000201:enzymology*; D050918:Phosphatidylethanolamine N-Methyltransferase; D010714:Phosphatidylethanolamines / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values
D010714:Phosphatidylethanolamines; D008715:Methionine; D008780:Methyltransferases; D050918:Phosphatidylethanolamine N-Methyltransferase; D002794:Choline