In vitro mass: activity distribution of lecithin--cholesterol acyltransferase among human plasma lipoproteins.
In order to compare the mass-activity distribution of lecithin-cholesterol acyltransferase (LCAT) among plasma lipoproteins separated by various ultracentrifugal or chromatographic procedures, we have quantified the enzyme by an electroimmunoassay technique using a specific antibody raised in the rabbit. This antibody, when added to whole serum, inhibited all of the enzyme activity present in it. The percent mass distribution of the enzyme among the lipoproteins isolated by rate-zonal ultracentrifugation (d 1.00-1.36 g/ml, SW 40 rotor, 37 000 rpm, 16 h) was as follows: very low density lipoproteins (VLDL), 0; low density lipoproteins (LDL), 6.2; HDL2, 6.5; HDL3, 12 and d greater than 1.21 g/ml fraction, 75. Measurement of LCAT activity of each lipoprotein fraction against mixed single bilayer lecithin-cholesterol vesicles (molar ratio, 4:1) containing apo A-I, indicated that VLDL, LDL and HDL2 were inactive or minimally active under the experimental conditions used, whereas HDL3 and the d greater than 1.21 g/ml fraction contained 17.5 and 79.9% of the total enzyme activity. Prolonged ultracentrifugation of the LCAT-containing lipoproteins resulted in the recovery of activity in the lipoprotein-free infranatant. In studies with lipoproteins linked to Sepharose 4B, LCAT was found to bind LDL, HDL2, and HDL3. It is concluded that LCAT is present in all the major lipoproteins except for VLDL. The activity appears to be dependent, at least in part, on the type of lipoproteins to which the enzyme is associated with.
D002500:Centrifugation, Isopycnic; D006801:Humans; D007106:Immune Sera; D008074:Lipoproteins / Q000097:blood*; D008077:Lipoproteins, LDL / Q000097:blood; D008079:Lipoproteins, VLDL / Q000097:blood; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000097:blood*
D007106:Immune Sera; D008074:Lipoproteins; D008077:Lipoproteins, LDL; D008079:Lipoproteins, VLDL; D007862:Phosphatidylcholine-Sterol O-Acyltransferase