Preparation of high-density lipoproteins labelled exclusively at the cholesteryl ester moiety.
The in vitro preparation of HDL radiolabelled at the cholesteryl ester moiety was determined by incubating different serum fractions with [3H]cholesteryl linoleate or cholesteryl [14C]oleate dissolved in dimethylsulfoxide (DMSO) and subsequent re-isolation of HDL from the incubation mixture by ultracentrifugation. Using the supernatant obtained by precipitation of apolipoprotein B-containing lipoproteins with phosphotungstate/MgCl2 or using the greater than 1.075 kg/l fraction of sera, the bulk of the added radioactivity could be recovered in the re-isolated HDL fraction, as demonstrated by density gradient ultracentrifugation or by gel chromatography. Incubation experiments using whole sera or the greater than 1.063 kg/l fraction of sera resulted in the preferential association of the labelled cholesteryl linoleate with apolipoprotein B-containing lipoproteins.
D001053:Apolipoproteins; D001055:Apolipoproteins B; D002250:Carbon Radioisotopes; D002499:Centrifugation, Density Gradient; D011232:Chemical Precipitation; D002788:Cholesterol Esters; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D008075:Lipoproteins, HDL / Q000302:isolation & purification; D008297:Male; D019301:Oleic Acid; D009829:Oleic Acids; D014316:Tritium
D001053:Apolipoproteins; D001055:Apolipoproteins B; D002250:Carbon Radioisotopes; D002788:Cholesterol Esters; D008075:Lipoproteins, HDL; D009829:Oleic Acids; D014316:Tritium; D019301:Oleic Acid; C021976:cholesteryl linoleate