Studies on the cofactor requirement for the acylation and hydrolysis reactions catalyzed by purified lecithin-cholesterol acyltransferase. Effect of low density lipoproteins and apolipoprotein A-i.
Because lecithin-cholesterol acyltransferase (LCAT) has been shown to carry out acylation of lysolecithin as well as hydrolysis of lecithin in addition to an esterification of cholesterol, the cofactor requirements of the three reactions catalyzed by the enzyme were studied. The purified enzyme required apolipoprotein A-I (apo A-I) for both the phospholipase A2 activity (release of free fatty acids from lecithin) and cholesterol esterification, whereas, low density lipoprotein (LDL) was required for the acylation of lysolecithin. Apo A-I and lecithin liposomes could not substitute for LDL for the activation of lysolecithin acyltransferase activity. Removal of apo A-I from the LDL preparation by affinity chromatography did not affect the activation of lysolecithin acylation, indicating that the contaminating apo A-I is not responsible for the activation. LDL facilitates cholesterol esterification in presence of labelled lecithin liposomes by providing the unesterified cholesterol. Removal of contaminating apo A-I, however, abolishes this LCAT activity which could be restored by addition of pure apo A-I. Lysolecithin inhibits both phospholipase A2 and LCAT activities, but LDL appeared to attenuate the effects of lysolecithin, in addition to stimulating the acylation of lysolecithin. These results show that apo A-I is not obligatory for all the reactions carried out by the enzyme, and that LDL plays an important role in the regulation of the hydrolysis and acylation reaction carried out by the enzyme.
D008243:1-Acylglycerophosphocholine O-Acyltransferase / Q000378:metabolism; D000215:Acylation; D016632:Apolipoprotein A-I; D001053:Apolipoproteins / Q000494:pharmacology*; D001054:Apolipoproteins A; D002384:Catalysis; D006801:Humans; D006868:Hydrolysis; D008077:Lipoproteins, LDL / Q000494:pharmacology*; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000378:metabolism / Q000494:pharmacology*; D010741:Phospholipases A; D054467:Phospholipases A2; D012709:Serum Albumin / Q000494:pharmacology
D016632:Apolipoprotein A-I; D001053:Apolipoproteins; D001054:Apolipoproteins A; D008077:Lipoproteins, LDL; D008244:Lysophosphatidylcholines; D012709:Serum Albumin; D008243:1-Acylglycerophosphocholine O-Acyltransferase; D007862:Phosphatidylcholine-Sterol O-Acyltransferase; D010741:Phospholipases A; D054467:Phospholipases A2