title: Synthetic amphipathic helical peptides that mimic apolipoprotein A-I in clearing cellular cholesterol.
abstract: Clearance of excess cholesterol from cells by HDL is facilitated by the interaction of HDL apolipoproteins with cell-surface binding sites or receptors, a process that may be important in preventing atherosclerosis. In this study, synthetic peptides containing 18-mer amphipathic helices of the class found in HDL apolipoproteins (class A) were tested for their abilities to remove cholesterol and phospholipid from cultured sterol-laden fibroblasts and macrophages and to interact with cell-surface HDL binding sites. Lipid-free peptides containing two identical tandem repeats of class A amphipathic helices promoted cholesterol and phospholipid efflux from cells and depleted cellular cholesterol accessible for esterification by acyl CoA/cholesterol acyltransferase, similar to what was observed for purified apolipoprotein A-I. Peptide-mediated removal of plasma membrane cholesterol and depletion of acyl CoA/cholesterol acyltransferase-accessible cholesterol appeared to occur by separate mechanisms, as the latter process was less dependent on extracellular phospholipid. The dimeric amphipathic helical peptides also competed for high-affinity HDL binding sites on cholesterol-loaded fibroblasts and displayed saturable high-affinity binding to the cell surface. In contrast, peptides with a single helix had little or no ability to remove cellular cholesterol and phospholipid, or to interact with HDL binding sites, suggesting that cooperativity between two or more helical repeats is required for these activities. Thus, synthetic peptides comprising dimers of a structural motif common to exchangeable apolipoproteins can mimic apolipoprotein A-I in both binding to putative cell-surface receptors and clearing cholesterol from cells.
authors.affiliation: Department of Medicine RG-26, University of Washington, Seattle 98195.
authors.forename: A J
authors.initials: AJ
authors.lastname: Mendez
authors.forename: G M
authors.initials: GM
authors.lastname: Anantharamaiah
authors.forename: J P
authors.initials: JP
authors.lastname: Segrest
authors.forename: J F
authors.initials: JF
authors.lastname: Oram
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016632:Apolipoprotein A-I / Q000378:metabolism*; D001667:Binding, Competitive; D002352:Carrier Proteins; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism*; D015256:Coenzyme A-Transferases / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008075:Lipoproteins, HDL / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D017433:Protein Structure, Secondary; D016601:RNA-Binding Proteins; D018110:Receptors, Lipoprotein / Q000378:metabolism; D012867:Skin / Q000166:cytology; D002785:Sterol O-Acyltransferase / Q000378:metabolism
chemical_list: D016632:Apolipoprotein A-I; D002352:Carrier Proteins; D008075:Lipoproteins, HDL; D010455:Peptides; D010743:Phospholipids; D016601:RNA-Binding Proteins; D018110:Receptors, Lipoprotein; C031616:high density lipoprotein receptors; C075197:high density lipoprotein binding protein; D002784:Cholesterol; D002785:Sterol O-Acyltransferase; D015256:Coenzyme A-Transferases; C031212:acyl-CoA transferase