Cholesterol-induced fluid-phase immiscibility in membranes.

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Abstract
The fluid-phase behavior of binary mixtures of cholesterol with phosphatidylcholines is investigated using magnetic resonance methods. Phospholipid biradicals provide the electron spin resonance spectroscopic resolution of two immiscible fluid phases in the dipalmitoylphosphatidylcholine-cholesterol system. Isotropic chemical shifts of the phospholipid carbonyl carbons in binary mixtures with cholesterol measured using solid-state high-resolution nuclear magnetic resonance methods furnish evidence for a putative hydrogen bond between the 3 beta-hydroxyl of cholesterol and the sn-2 carbonyl of the phospholipid. The location in the bilayer of cholesterol in the two fluid phases is determined by measuring spin label-enhanced spin-lattice relaxation rates of the 13C nuclei of both the phospholipid and cholesterol molecules. These results suggest, in a time-averaged sense, that in the cholesterol-poor fluid phase the cholesterol molecule essentially spans the bilayer, whereas in the cholesterol-rich fluid phase the molecule is present in both monolayers of the bilayer.