Phosphatidylethanolamine:dolichol acyltransferase. Characterization and partial purification of a novel rat liver enzyme.
Incubation of rat or human post-heparin plasma with [3H]dolichol incorporated in liposomes consisting of dioleoyl phosphatidylcholine:dioleoyl phosphatidylethanolamine (3:1) resulted in the formation of radioactive dolichyl oleate. Non-heparinized plasma did not esterify dolichol, and, hence, the enzyme involved is probably associated with the cell surface and released into the blood by heparin. The major location of this activity was the liver, and, therefore, a partial purification of the enzyme from heparinized rat liver perfusates was performed using DEAE-Sephacel and heparin-Sepharose chromatography. The dolichol acyltransferase activity copurified with hepatic lipase activity in a lipid-protein complex of 350 kDa. Optimal acylation is achieved at pH 7.5 in the presence of 5% plasma and 20 mM Ca2+. Esterification can only be obtained when dolichol is present in a phospholipid bilayer, and the reaction is strongly stimulated by unsaturated phosphatidylethanolamine or phosphatidylserine. Radiolabeling experiments demonstrated that the primary acyl donor is phosphatidylethanolamine from which the fatty acid is transferred exclusively from position 1. Neither cholesterol nor retinol are esterified by the enzyme, and the reaction is not stimulated by acyl-CoA. Both the extracellular localization and the mechanism of transacylation clearly distinguish this new enzyme from the acyl-CoA:dolichol acyltransferase described earlier in microsomes.
Sindelar
P
P
Clinical Research Centre, Huddinge Hospital, Karolinska Institutet, Stockholm, Sweden.
Chojnacki
T
T
Valtersson
C
C
D000217:Acyltransferases / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D002413:Cations, Divalent / Q000494:pharmacology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006801:Humans; D007527:Isoenzymes / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008049:Lipase / Q000378:metabolism; D008081:Liposomes; D008099:Liver / Q000201:enzymology*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology; D008970:Molecular Weight; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity
D002413:Cations, Divalent; D007527:Isoenzymes; D008081:Liposomes; D000217:Acyltransferases; C077400:phosphatidylethanolamine - dolichol acyltransferase; D008049:Lipase