Cholesterol efflux from macrophages mediated by high-density lipoprotein subfractions, which differ principally in apolipoprotein A-I and apolipoprotein A-II ratios.
High-density lipoprotein (HDL) was fractionated by preparative isoelectric focussing into six distinct subpopulations. The major difference between the subfractions was in the molar ratio of apolipoprotein A-I to apolipoprotein A-II, ranging from 2.1 to 0.5. The least acidic particles had little apolipoprotein A-II, were larger and contained the most lipid. The efflux capacity of the HDL subfractions was tested with mouse peritoneal macrophages and a mouse macrophage cell line (P388D1), either fed with acetylated low-density lipoprotein or free cholesterol. All the HDL subfractions were equally able to efflux cholesterol. The efflux was concentration dependant and linear for the first 6 h. The HDL subfractions bound with high affinity (Kd = 6.7-7.9 micrograms/ml) at 4 degrees C to the cell surface of P388D1 cells (211,000-359,000 sites/cell). Ligand blotting showed that all the HDL subfractions bound to membrane polypeptides at 60, 100, and 210 kDa. These HDL binding proteins may represent HDL receptors. In summary HDL particles, which differed principally in ratio of apolipoprotein A-I to apolipoprotein A-II behaved in a similar manner for both cholesterol efflux and cell surface binding.
University Hospital, Department of Cardiology, University of Heidelberg, Germany.
E
E
von Hodenberg
S
S
Heinen
K E
KE
Howell
C
C
Luley
W
W
Kübler
H M
HM
Bond
D000818:Animals; D016632:Apolipoprotein A-I / Q000032:analysis* / Q000378:metabolism / Q000494:pharmacology; D016633:Apolipoprotein A-II / Q000032:analysis* / Q000378:metabolism / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism*; D002788:Cholesterol Esters / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D006801:Humans; D007525:Isoelectric Focusing; D007700:Kinetics; D008075:Lipoproteins, HDL / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D053440:Lipoproteins, HDL3; D008077:Lipoproteins, LDL / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D010529:Peritoneal Cavity / Q000166:cytology
D016632:Apolipoprotein A-I; D016633:Apolipoprotein A-II; D002788:Cholesterol Esters; D008075:Lipoproteins, HDL; D053440:Lipoproteins, HDL3; D008077:Lipoproteins, LDL; C032629:acetyl-LDL; D002784:Cholesterol; D004229:Dithiothreitol