Phase structures of binary lipid bilayers as revealed by permeability of small molecules.

The effects of changes in bilayer phase structure on the permeability of acetic acid and trimethylacetic acid were studied in large unilamellar vesicles (LUVs) composed of dipalmitoylphosphatidylcholine (DPPC)/cholesterol (CHOL), dihexadecylphosphatidylcholine (DHPC)/CHOL, or DPPC/dimyristoylphosphatidylcholine (DMPC) using an NMR line-broadening method. Phase transitions were induced by changes in temperature and lipid composition (i.e., XCHOL was varied from 0.0 to 0.5 and XDMPC from 0.0 to 1.0). In DPPC/CHOL and DHPC/CHOL bilayers, the addition of CHOL induces only a modest change in the permeability coefficient (Pm) of acetic acid in the gel-phase (Pbeta') but significantly reduces Pm in ordered and disordered liquid-crystalline phases (Lo and Lalpha). Abrupt changes in slopes in semi-logarithmic plots of Pm vs. XCHOL occur at specific values of XCHOL and temperature corresponding to the boundaries between Pbeta' and Lo or between Lalpha and Lo phases. In most respects, phase diagrams generated from the break points in plots of Pm vs. XCHOL obtained at various temperatures in DHPC/CHOL and DPPC/CHOL bilayers closely resemble those constructed previously for DPPC/CHOL bilayers using NMR and DSC methods. Above Tm, the phase diagrams generated from permeability data reveal the presence of both the disordered (Lalpha) and the ordered (Lo) liquid-crystalline phases, as well as the two-phase coexistence region. In DPPC/DMPC bilayers, the addition of DMPC increases Pm dramatically in the gel phase but only slightly in the liquid-crystalline phase. Abrupt changes in slopes in semi-logarithmic plots of Pm vs. XDMPC also occur at specific values of XDMPC and temperature, from which a phase diagram can be constructed which closely resembles diagrams obtained previously by other methods. These correlations indicate that trans-bilayer permeability measurements can be used to construct lipid bilayer phase diagrams. Positive deviations of Pm from predicted values based on the phase lever rule are observed in the two-phase coexistence regions with the degree of the deviation depending on bilayer chemical composition and temperature. These results may reflect a specific contribution of the interfacial region between two phases to higher solute permeability or may be due to the higher lateral compressibility of lipid bilayers in the two-phase coexistence region. Copyright 1998 Elsevier Science B.V.