The ability of multilamellar vesicles of phosphatidylcholine and phosphatidylethanolamine in aqueous phase to prevent access to cholesterol by a nonpolar solvent was examined. Phosphatidylethanolamine vesicles. In mixed vesicles, cholesterol was retained in proportion to the amount of phosphatidylcholine. To alter the charge and hydration of head groups, pH was adjusted from 1.2 to 12.5. Above pH 8, both phosphatidylethanolamine and phosphatidylcholine retained sterol in a 1:1 molar ratio of phospholipid to cholesterol, regardless of acyl side chain composition. Between pH 2.0 and pH 8.0, sterol retention varied with type of head group and side chain. Lipids with 16-carbon saturated side chains retained more sterol than 18-carbon unsaturated or 12-carbon saturated side chains. Between pH 1.1 and 2.0, none of the phosphatidylethanolamines retained sterol, but long chain phosphatidylcholines, saturated or unsaturated, retained sterol in a 1:1 molar ratio of phospholipid to sterol. Short chain phosphatidylethanolamines and phosphatidylcholines retained 0 to 20% at the low- to mid-pH range. Size of multilamellar vesicles, measured by Doppler effect light scattering analysis, had no bearing on sterol retention. Sonication of vesicles, which increases surface curvature, increases the retention of sterol. Fluorescence polarization indicated that cholesterol does not interact with DPPC or DLPC side chains. The observations can be interpreted in terms of space requirements of head groups, including charge repulsion and hydration. Other factors, such as monovalent cation replacement by protons, juxtaposition of charged groups on vesicle surfaces and length and unsaturation of acyl side chains affect the affinity of phospholipids for cholesterol.
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