N-myristoylated phosphatidylethanolamine: Interfacial behavior and interaction with cholesterol

The interfacial packing behavior of N-myristoyldimyristoylphosphatidylethanolamine (N-14:0 DMPE) and its interaction with cholesterol were characterized and compared to the behavior of dimyristoylphosphatidylethanolamine (DMPE) using an automated Langmuir type film balance. Surface pressure and surface potential were monitored as a function of lipid cross-sectional molecular area. N-14:0 DMPE exhibited two-dimensional (2D) phase transitions of a liquid-expanded to condensed nature at many temperatures in the 15-30 °C range, but isotherms showed only condensed behavior at 15 °C. The sharp decline in the surface compressional moduli upon entering the 2D-transition region is consistent with differences in the partial molar areas of coexisting liquid-expanded (chain-disordered) and condensed (chain-ordered) phases. Including Ca²⁺ in the subphase beneath the negatively charged N-14:0 DMPE caused a downward shift in the 2D-transition onset pressure even in the presence of 100 mM NaCl. The average dipole moments perpendicular to the lipid-water interface for N-14:0 DMPE's liquid-expanded and condensed phases were higher than those of DMPE. At surface pressures sufficiently low (<10 mN/m) to produce liquid-expanded phase behavior in pure N-14:0 DMPE, mixing with cholesterol resulted in a classic "condensing effect". Maximal area condensation was observed near equimolar N-14:0 DMPE/ cholesterol. Insights into mixing behavior at high surface pressures that mimic the lipid cross-sectional areas of biomembranes were provided by analyzing the surface compressional moduli as a function of cholesterol mole fraction. Complex mixing patterns were observed that deviated significantly from theoretical ideal mixing behavior suggesting the presence of lipid "complexes" and/or a liquid-ordered phase at high sterol mole fractions (>0.35) and low to intermediate surface pressures (<20 mN/m) as well as the possible coexistence of relatively immiscible solid phases at higher surface pressures (e.g., 35 mN/m).