Isolation and specificity of rat lecithin : Cholesterol acyltransferase: Comparison with the human enzyme using reassembled high-density lipoproteins containing ether analogs of phosphatidylcholine

Abstract

Rat plasma lecithin:cholesterol acyltransferase, a 68 kDa glycoprotein, has been purified 14000-fold by a modification of a procedure used for the human enzyme. The activity of lecithin : cholesteryl acyltransferase in human and rat plasma are the same, although activation of both enzymes by human apolipoprotein A-I is greater than that produced by rat apolipoprotein A-I. Using reassembled high-density lipoproteins composed of human apolipoprotein A-I, phosphatidylcholine ethers and a series of different phosphatidylcholines, the separate effects of molecular species specificity and microenvironment on the rate of cholesteryl ester formation was determined. Substitution of a fluid lipid, 1-palmityl-2-oleyl- sn-glvcero-3-phosphonlcholine. for a solid lipid, 1,2-dipalmityl- sn-glycero-3-phosphorylcholine, produced an 8-fold increase in the activity of all molecular species of phosphatidylcholine. With either solid or fluid lipid environments, the activity decreased as a function of increasing chain length of saturated acyl groups. Addition of one or more double bonds greatly increased the activity of a given saturated homologue. One major difference between the molecular specificity of rat and human lecithin:cholesteryl acyltransferase was that the latter had a two-fold preference for phosphatidylcholines containing arachidonate at the sn-2-position.