title: Role of lecithin:cholesterol acyltransferase and apolipoprotein A-I in cholesterol esterification in lipoprotein-X in vitro.
abstract: Lipoprotein-X (Lp-X) is an abnormal particle present in the plasma of patients with familial lecithin:cholesterol acyltransferase (LCAT) deficiency syndromes or cholestatic liver disease. Compared to other lipoproteins, Lp-X contains a high content of unesterified cholesterol (30%, w/w) to phosphatidylcholine (60%, w/w). The objective of this study was to evaluate the role of LCAT and apolipoprotein A-I (apoA-I) in Lp-X metabolism in vitro and to elucidate the regulation of cholesterol esterification in this unique lipoprotein. Lp-X isolated from sera of patients with obstructive jaundice had a high content of unesterified cholesterol and phosphatidylcholine and contained apolipoprotein E, apoCs, and albumin. Although human recombinant LCAT used as an enzyme source did bind to isolated Lp-X, no cholesterol esterification was detected. However, addition of human apoA-I in the presence of albumin resulted in significant cholesterol esterification in Lp-X (Vmax 0.25 +/- 0.04 nmol/h per microgram LCAT protein). Exogenous apoA-I did not change the size of Lp-X particle as determined by quasi-elastic light scattering analysis. A reduction in Lp-X size was observed when both apoA-I and LCAT were included in the reaction mixture (from 47 nm to 42 nm). Furthermore, addition of apoA-I (but not HDL) dramatically changed the electrophoretic mobility of Lp-X from cathodic to anodic migration. Such changes are not due to displacement of apoC or apoE proteins from Lp-X by apoA-I. While increasing apoA-I concentration (up to 35 micrograms/ml) in the reaction mixture stimulated cholesterol esterification in Lp-X, addition of apoA-I at the concentration of 8 micrograms/ml inhibited cholesterol esterification in VLDL, LDL, and HDL by more than 50%. Albumin was required for the LCAT reaction to Lp-X. Our results suggest that while LCAT binds to isolated Lp-X, apoA-I is needed for the LCAT reaction to proceed. The presence of apoA-I does not result in the displacement of apoCs and apoE from Lp-X and addition of apoA-I changes the electrophoretic mobility but not the size of Lp-X.
authors.affiliation: Department of Pathology, University of British Columbia, Vancouver, Canada.
authors.forename: K
authors.initials: K
authors.lastname: O
authors.forename: J
authors.initials: J
authors.lastname: Frohlich
mesh_terms: D000818:Animals; D016632:Apolipoprotein A-I / Q000502:physiology*; D002460:Cell Line; D002780:Cholestasis, Intrahepatic / Q000097:blood* / Q000201:enzymology; D002788:Cholesterol Esters / Q000097:blood*; D006224:Cricetinae; D004951:Esterification; D006801:Humans; D007863:Lecithin Cholesterol Acyltransferase Deficiency / Q000097:blood* / Q000201:enzymology; D008073:Lipoprotein-X / Q000097:blood* / Q000302:isolation & purification; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000502:physiology*
chemical_list: D016632:Apolipoprotein A-I; D002788:Cholesterol Esters; D008073:Lipoprotein-X; D007862:Phosphatidylcholine-Sterol O-Acyltransferase