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Biophys J, May 2000, p. 2441-2451, Vol. 78, No. 5

A 2H NMR Study of Macroscopically Aligned Bilayer Membranes Containing Interfacial Hydroxyl Residues

Volker Kurze, Bernhard Steinbauer, Thomas Huber, and Klaus Beyer

Adolf-Butenandt-Institut, Lehrstuhl für Stoffwechselbiochemie der Universität München, 80336 München, Germany

The polar interface of membranes containing phosphatidylglycerol or cholesterol was studied by 2H nuclear magnetic resonance (NMR) as a function of membrane hydration. The membranes were macroscopically aligned and hydrated with deuterium oxide. Water uptake and membrane annealing was achieved under NMR control, using a novel hydration technique. Well-resolved 2H quadrupolar doublets were obtained from individual hydroxyl residues and from the interlamellar water. The response of the phosphatidylglycerol headgroup and of the cholesterol molecule to the spontaneous evaporation of interlamellar water could be thus monitored continuously. It is shown that the phosphatidylglycerol headgroup undergoes changes of conformation and average orientation with respect to the membrane surface and that the off-axis motion of the cholesterol molecule decreases. The deuteron exchange between hydroxyl residues and surface-associated D2O was determined by an inversion transfer technique. The exchange rates of the hydroxyl residues in the phosphatidylglycerol headgroup were different and depended strongly on the total hydration of the membrane. Significantly lower and almost hydration-independent rates were obtained for cholesterol. These results will be discussed with reference to earlier reports on the headgroup dynamics of phosphatidylglycerol and on the interaction of cholesterol with the membrane-water interface.

Biophys J, May 2000, p. 2441-2451, Vol. 78, No. 5
© 2000 by the Biophysical Society   0006-3495/00/05/2441/11  $2.00


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