title: Depletion with Cyclodextrin Reveals Two Populations of Cholesterol in Model Lipid Membranes.
abstract: Recent results provide evidence that cholesterol is highly accessible for removal from both cell and model membranes above a threshold concentration that varies with membrane composition. Here we measured the rate at which methyl-β-cyclodextrin depletes cholesterol from a supported lipid bilayer as a function of cholesterol mole fraction. We formed supported bilayers from two-component mixtures of cholesterol and a PC (phosphatidylcholine) lipid, and we directly visualized the rate of decrease in area of the bilayers with fluorescence microscopy. Our technique yields the accessibility of cholesterol over a wide range of concentrations (30-66 mol %) for many individual bilayers, enabling fast acquisition of replicate data. We found that the bilayers contain two populations of cholesterol, one with low surface accessibility and the other with high accessibility. A larger fraction of the total membrane cholesterol appears in the more accessible population when the acyl chains of the PC-lipid tails are more unsaturated. Our findings are most consistent with the predictions of the condensed-complex and cholesterol bilayer domain models of cholesterol-phospholipid interactions in lipid membranes.
authors: Department of Chemistry, University of Washington, Seattle, Washington.
authors: Jonathan P
authors: JP
authors: Litz
authors: Department of Chemistry, University of Washington, Seattle, Washington; Department of Applied Mathematics, University of Washington, Seattle, Washington.
authors: Niket
authors: N
authors: Thakkar
authors: Department of Chemistry, University of Washington, Seattle, Washington.
authors: Thomas
authors: T
authors: Portet
authors: Department of Chemistry, University of Washington, Seattle, Washington; Department of Physics, University of Washington, Seattle, Washington. Electronic address: slkeller@chem.washington.edu.
authors: Sarah L
authors: SL
authors: Keller
mesh_terms: D002784:Cholesterol / Q000737:chemistry*; D003505:Cyclodextrins / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry
chemical_list: D003505:Cyclodextrins; D008051:Lipid Bilayers; D008081:Liposomes; D002784:Cholesterol