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Hydration and Lateral Organization in Phospholipid Bilayers Containing Sphingomyelin: A ²H-NMR Study

Lehrstuhl für Stoffwechselbiochemie der Universität München, Munich, Germany

Correspondence: Address reprint requests to Dr. Klaus Beyer, Lehrstuhl für Stoffwechselbiochemie der Universität München, Schillerstr. 44, 80336 München, Germany. Tel.: 49-89-599-6470; Fax: 49-89-599-6415; E-mail: kbeyer{at}med.uni-muenchen.de.

Interfacial properties of lipid bilayers were studied by ²H nuclear magnetic resonance spectroscopy, with emphasis on a comparison between phosphatidylcholine and sphingomyelin. Spectral resolution and sensitivity was improved by macroscopic membrane alignment. The motionally averaged quadrupolar interaction of interlamellar deuterium oxide was employed to probe the interfacial polarity of the membranes. The D₂O quadrupolar splittings indicated that the sphingomyelin lipid-water interface is less polar above the phase transition temperature T_m than below T_m. The opposite behavior was found in phosphatidylcholine bilayers. Macroscopically aligned sphingomyelin bilayers also furnished ²H-signals from the amide residue and from the hydroxyl group of the sphingosine moiety. The rate of water-hydroxyl deuteron exchange could be measured, whereas the exchange of the amide deuteron was too slow for the inversion-transfer technique employed, suggesting that the amide residue is involved in intermolecular hydrogen bonding. Order parameter profiles in mixtures of sphingomyelin and chain-perdeuterated phosphatidylcholine revealed an ordering effect as a result of the highly saturated chains of the sphingolipids. The temperature dependence of the ²H quadrupolar splittings was indicative of lateral phase separation in the mixed systems. The results are discussed with regard to interfacial structure and lateral organization in sphingomyelin-containing biomembranes.

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