Bicellar dispersions of chain perdeuterated 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC-d54), 1,2-dimyristoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DMPG), and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC), with molar ratios of 3:1:1, were studied using variable-pressure (2)H NMR spectroscopy at hydrostatic pressures up to 125 MPa. Upon warming of the dispersions, spectra at ambient pressure indicated a progressive coalescence from small bilayered disks undergoing isotropic reorientation to more extended micellar structures in which spectra indicated anisotropic reorientation and, under some conditions, magnetic orientation and finally to randomly oriented lamellae or multilamellar vesicles. Temperatures for the onsets of anisotropic reorientation and random lamellar orientation increased with pressure at rates of 0.22 and 0.15 °C/MPa, respectively. In the 3.5-T magnetic field used for this work, magnetic orientation within the intermediate phase was not observed at 83 MPa or higher pressures. Comparison of spectra obtained at fixed pressure showed significant asymmetry between behaviors upon warming and cooling. For samples of DMPC-d54/DMPG/DHPC (3:1:1), but not DMPC-d54/DHPC (4:1), a persistent interdigitated phase was formed after repeated cooling from high temperature at 83 MPa. This is likely a metastable phase and might reflect kinetic trapping of the short-chain lipid component, DHPC, in a nonequilibrium spatial distribution as temperature is lowered at high pressure. Bicellar dispersions typically behave differently upon warming and cooling, and these observations could provide some insight into the observed behaviors in such systems. This work also suggests the possibility of trapping bicellar dispersions in persistent nonequilibrium morphologies.

Bicellar dispersions of chain perdeuterated 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC-d54) and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) were prepared with the molar fraction of DHPC held fixed at 20% and varying amounts of DMPC replaced by the anionic lipid 1,2-dimyristoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DMPG). (2)H NMR spectra were examined to assess the effect of added DMPG on mixture phase behavior and morphology. Quadrupole echo decay and quadrupole-Carr-Purcell-Mieboom-Gill echo train measurements provided information about slow motions contributing to echo decay in the high temperature phases. The spectra and quadrupole echo decay properties of DMPC-d54/DHPC (4:1) and DMPC-d54/DMPG/DHPC (3:1:1) were qualitatively similar. With increasing DMPG concentration, the transition between the magnetically orientable phase and the higher temperature phase became increasingly distinct, and the spectral shape and echo decay characteristics of the high temperature bicellar phase became increasingly similar to those of DMPC-d54 in the liquid crystalline phase. The observation that DMPG changes spectra in the orientable phase incrementally while increasing the distinction between the orientable and high temperature bicellar phases provides new insights into how DMPG influences bicellar mixture morphology.

Dispersions of lipid mixtures comprising long- and short-chain phospholipids (bicellar mixtures) can form small isotropically reorienting particles (bilayered micelles), magnetically orientable stuctures, or unorientable lamellar structures. Application of hydrostatic pressure can also induce interdigitation of the long-chain lipid components. In this work, variable-pressure H NMR was used to study the effect of head group charge on the barotropic behavior of bicellar mixtures. Observations at pressures up to 152 MPa and temperatures up to 64 °C were combined with earlier observations at lower pressure and lower temperature to obtain a pressure-temperature phase diagram for DMPC-d/DMPG/DHPC (3:1:1). In this phase diagram, a region corresponding to small, isotropically reorienting particles at lower temperature and higher pressure is separated from a region corresponding to unorientable lamellar organization, at higher temperature and lower pressure, by a band in which the magnetically orientable phase is stable below ∼100 MPa and in which an interdigitated gel phase is stable above ∼120 MPa. From ∼46 to ∼52 °C, the dispersion transforms directly from the unorientable lamellar to isotropically reorienting particle phases upon isothermal pressurization. The extent to which this behavior reflects the presence of anionic lipid in the long-chain fraction of this mixture is illustrated by comparison with spectral series obtained during isothermal pressurization of DMPC-d/DHPC (4:1) and DMPC-d/DMPG/DHPC (2.7:1.3:1) at selected temperatures. These observations show how electrostatic interactions at a bilayer surface can affect the balance between hydrophobic and hydrophilic interactions that is reflected by a dispersion's barotropic phase behavior.