title: Specific phospholipid association with apolipoprotein A-I stimulates cholesterol efflux from human fibroblasts. Studies with reconstituted sonicated lipoproteins.
abstract: To understand how the lipid composition of high density lipoprotein mediates the efflux of cellular cholesterol, we have characterized the effects of variations in the lipid composition of well defined model sonicated apolipoprotein A-I (apoA-I)-containing lipoprotein (LpA-I) particle on cholesterol efflux from cultured human skin fibroblasts. LpA-I particles with varying content of phosphatidylcholine (POPC), phosphatidylinositol, sphingomyelin, cholesterol ester, and triolein were prepared by co-sonication. Association of as little as 5 mol of phosphatidylcholine with apoA-I is sufficient to transform lipid-free apoA-I into a distinct lipoprotein-like particle that is a significantly better acceptor of cellular cholesterol. Increasing the ratio of POPC/apoA-I from 5/1 to 35.5/1 in the sonicated LpA-I is associated with a significant increase in the release of cellular cholesterol. At low POPC/apoA-I ratios, native gradient gel electrophoresis of the LpA-I shows these lipoproteins to be small complexes (around 5-6 nm), with only 1 molecule of apoA-I (Lp1A-I). At a POPC/apoA-I ratio above 11/1, LpA-I form well defined complexes that contain 2 molecules of apoA-I (Lp2A-I) and range in size from 7.6 to 7.7 nm. Inclusion of sphingomyelin into an Lp1A-I further stimulates cholesterol efflux significantly. In contrast, inclusion of either sphingomyelin or phosphatidylinositol into a sonicated Lp2A-I has no effect on cholesterol efflux. Incorporation of cholesterol ester and/or triolein into an Lp2A-I particle is associated with a small reduction in cholesterol efflux to these lipoproteins. Therefore, cholesterol efflux from human fibroblasts is directly proportional to the amount and type of phospholipid in a sonicated LpA-I particle. Changes in the conformation and charge of apoA-I that result from changes in the lipid composition of a sonicated LpA-I particle appear to directly affect the ability of the lipoprotein to bind and retain cholesterol molecules. These data therefore suggest that the adsorption/desorption of cholesterol molecules to/from a sonicated LpA-I complex may be less sensitive to interfacial lipid-lipid interactions, but may depend on a conformation-dependent ability of apoA-I to bind cholesterol.
authors.affiliation: Lipoprotein and Atherosclerosis Group, University of Ottawa Heart Institute, University of Ottawa, Ottawa, Ontario K1Y 4E9, Canada.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhao
authors.forename: D L
authors.initials: DL
authors.lastname: Sparks
authors.forename: Y L
authors.initials: YL
authors.lastname: Marcel
mesh_terms: D016632:Apolipoprotein A-I / Q000097:blood / Q000302:isolation & purification / Q000494:pharmacology*; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism*; D002788:Cholesterol Esters / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D010713:Phosphatidylcholines / Q000494:pharmacology; D010716:Phosphatidylinositols / Q000494:pharmacology; D010743:Phospholipids / Q000494:pharmacology*; D012867:Skin / Q000378:metabolism; D013109:Sphingomyelins / Q000494:pharmacology; D013329:Structure-Activity Relationship; D014304:Triolein / Q000494:pharmacology; D014465:Ultrasonics
chemical_list: D016632:Apolipoprotein A-I; D002788:Cholesterol Esters; D010713:Phosphatidylcholines; D010716:Phosphatidylinositols; D010743:Phospholipids; D013109:Sphingomyelins; D014304:Triolein; D002784:Cholesterol; C028694:1-palmitoyl-2-oleoylphosphatidylcholine