Increased cholesterol esterification in rat liver microsomes in purified non-specific phospholipid transfer protein.
The effect of the non-specific phospholipid transfer protein purified from rat liver on the activity of acyl-CoA:cholesterol acyltransferase (EC 2.3.1.26) in rat liver microsomes was studied. The activity of cholesterol acyltransferase was measured from the rate of incorporation of [1-14C] oleoyl-CoA into cholesteryl oleate. Activity was stimulated by preincubation by the microsomes with the non-specific phospholipid transfer protein alone, but most effectively when vesicles consisting of phosphatidylcholine/cholesterol (molar ratio 1:1) also were present in the preincubation mixture. Preincubation with vesicles consisting of only phosphatidylcholine or phosphatidylcholine/phosphatidylethanolamine (molar ratio 1:1) had no effect. The stimulating effect is dependent on transfer protein and vesicle concentration and on the length of preincubation. Treatment of the transfer protein with N-ethylmaleimide abolished its effect on cholesterol ester formation. Preincubation of the microsomes with transfer protein and phosphatidylcholine/cholesterol vesicles containing radioactively labeled cholesterol shows that exogenous cholesterol is converted readily to cholesterol ester. The data are explained by the ability of non-specific phospholipid transfer protein to effect net transfer of cholesterol to those microsomes that contain cholesterol acyltransferase. Enlargement of the cholesterol substrate pool would then give rise to stimulation of the cholesterol acyltransferase activity. This study suggests a role for the transfer protein in modulating cholesterol metabolism by its ability to transport cholesterol between membranes.
D000214:Acyl Coenzyme A / Q000378:metabolism; D000818:Animals; D002250:Carbon Radioisotopes; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D002788:Cholesterol Esters / Q000096:biosynthesis*; D007700:Kinetics; D008297:Male; D008565:Membrane Proteins; D008862:Microsomes, Liver / Q000378:metabolism*; D049294:Phospholipid Transfer Proteins; D051381:Rats; D011919:Rats, Inbred Strains; D014316:Tritium
D000214:Acyl Coenzyme A; D002250:Carbon Radioisotopes; D002352:Carrier Proteins; D002788:Cholesterol Esters; D008565:Membrane Proteins; D049294:Phospholipid Transfer Proteins; D014316:Tritium; C017585:oleoyl-coenzyme A; D002784:Cholesterol