Biophysical Journal 61: 1025-1035 (1992)
© 1992 the Biophysical Society

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Changes of the physical properties of the liquid-ordered phase with temperature in binary mixtures of DPPC with cholesterol

A ²H-NMR, FT-IR, DSC, and neutron scattering study

Technische Universität München, Physik Department E22, D-W-8046 Garching, Germany

ABSTRACT

The structure of the so-called liquid-ordered (lo) phase of binary mixtures of DPPC-d₆₂ with cholesterol was studied between 20-50 mol% cholesterol using ²H-NMR, FT-IR, DSC, and neutron specular reflection. Different model systems such as multilamellar vesicles, spherical supported vesicles, and oriented multilayers were used. We observed significant changes of the lo phase structure in the physiological relevant temperature region between 30-45°C. ²H-NMR in combination with lineshape simulations provides evidence for a drastic effect of cholesterol on the shape of multilamellar vesicles due to magnetic field orientation. Moreover, the data indicates a significant change of the extent of this partial orientation for DPPC-d₆₂ multilamellar vesicles containing 25 mol% cholesterol between 36-42°C. The semiaxes ratio of the (due to magnetic field orientation) ellipsoidal multilamellar vesicles changes over this temperature range by 25%. ²H-NMR and FT-IR measurements indicate changes of the average orientational order at the bilayer center in the same temperature range and a significant increase of the number of end-gauche conformers while the majority of the methylene groups remain in an all-trans conformation. Additionally, specular reflection of neutrons shows a concomitant reduction of the bilayer thickness by 4 Å. Based on a model of the arrangement of DPPC and cholesterol in the lo phase, a molecular mechanism is proposed in which increasing the temperature between 30 and 45°C has the effect of driving cholesterol from the bilayer center towards the head group region.

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