Properties of acyl-CoA:cholesterol O-acyltransferase in aortic microsomes from atherosclerotic rabbits.

Acyl-CoA:cholesterol O-acyltransferase (EC 2.3.1.26) was in microsomal fractions from atherosclerotic rabbit aortic tissue. Activity was increased over 70-fold following 8-11 weeks of cholesterol feeding. Comparison of the lipid composition of aortic microsomal fractions from control and cholesterol-fed animals showed a 2-fold increase in the molar ratio of unesterified cholesterol to phospholipid in the cholesterol-fed group, although no change in phospholipid content or composition was found. Aortic microsomes were fractionated by sucrose density gradient centrifugation. Acyl-CoA:cholesterol O-acyltransferase was localized in fractions containing neutral alpha-glucoside activity but was clearly separated from 5'-nucleotidase activity. The cholesteryl ester formed during in vitro incubation of incubations of microsomes with either [1-14C]-oleoyl CoA or [7-3H]cholesterol was localized in the same region of the density gradient as acyl-CoA:cholesterol O-acyltransferase. The studies indicate that the increased acyl-CoA:cholesterol O-acyltransferase activity found in cells from aortic atherosclerotic lesions is due to both an increased amount of enzyme and to an expanded pool of microsomal cholesterol which is available for esterification.