Effect of cholesteryl ester on the distribution of fluorescent cholesterol analogues in triacylglycerol-rich emulsions.

Lipid emulsions consisting of a surface monolayer of phospholipid enclosing a core of neutral lipid (cholesteryl ester and/or triacylglycerol) are useful models of the lipid phase of lipoproteins. The physical state of the emulsion surface may determine the extent and nature of interaction of enzymes and lipid transfer proteins (e.g., lipoprotein lipase, cholesteryl ester transfer protein) with the particle. Unesterified cholesterol, which is a major determinant of the physical state of the surface phase, is able to partition between surface and core compartments. This report describes a fluorescence quenching method which determines the equilibrium distribution of a fluorescent cholesterol analogue (dehydroergosterol) between the surface and core compartments of triacylglycerol-rich emulsions. Quenching by iodide is used to distinguish a pool of unesterified cholesterol readily accessible to the aqueous phase. Quenching by 5-nitroxystearate identifies a pool of unesterified cholesterol in the phospholipid monolayer and the pool of unesterified cholesterol in the core compartment is found by difference. It is shown that the substitution of cholesteryl oleate for triolein in the core of the emulsion substantially increases the partition of unesterified cholesterol into the core compartment with a consequent depletion of unesterified cholesterol in the surface monolayer. The distribution of unesterified cholesterol between surface and core compartments is largely enthalpically driven.