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Biophys J, October 2002, p. 2053-2063, Vol. 83, No. 4

Properties of Mixtures of Cholesterol with Phosphatidylcholine or with Phosphatidylserine Studied by 13C Magic Angle Spinning Nuclear Magnetic Resonance

Richard M. Epand,* Alex D. Bain,dagger Brian G. Sayer,dagger Diana Bach,Dagger and Ellen Wachtel§

 *Department of Biochemistry and  dagger Department of Chemistry, McMaster University, Hamilton, Ontario L8S 4M1, Canada; and  Dagger Department of Biological Chemistry and  §Chemical Services Unit, Weizmann Institute of Science, Rehovot, Israel

The behavior of cholesterol is different in mixtures with phosphatidylcholine as compared with phosphatidylserine. In 13C cross polarization/magic angle spinning nuclear magnetic resonance spectra, resonance peaks of the vinylic carbons of cholesterol are a doublet in samples containing 0.3 or 0.5 mol fraction cholesterol with 1-palmitoyl-2-oleoyl phosphatidylserine (POPS) or in cholesterol monohydrate crystals, but a singlet with mixtures of cholesterol and 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC). At these molar fractions of cholesterol with POPS, resonances of the C-18 of cholesterol appear at the same chemical shifts as in pure cholesterol monohydrate crystals. These resonances do not appear in samples of POPS with 0.2 mol fraction cholesterol or with POPC up to 0.5 mol fraction cholesterol. In addition, there is another resonance from the cholesterol C18 that appears in all of the mixtures of phospholipid and cholesterol but not in pure cholesterol monohydrate crystals. Using direct polarization, the fraction of cholesterol present as crystallites in POPS with 0.5 mol fraction cholesterol is found to be 80%, whereas with the same mol fraction of cholesterol and POPC none of the cholesterol is crystalline. After many hours of incubation, cholesterol monohydrate crystals in POPS undergo a change that results in an increase in the intensity of certain resonances of cholesterol monohydrate in 13C cross polarization/magic angle spinning nuclear magnetic resonance, indicating a rigidification of the C and D rings of cholesterol but not other regions of the molecule.

Biophys J, October 2002, p. 2053-2063, Vol. 83, No. 4
© 2002 by the Biophysical Society   0006-3495/02/10/2053/11  $2.00


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