Permeability of liposomal membranes to water: Results from the magnetic field dependence of T1 of solvent protons in suspensions of vesicles with entrapped paramagnetic ions

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Abstract
The diffusive permeability to water molecules, Pd, of lipid vesicles with entrapped paramagnetic solute ions can be determined rapidly from analysis of the magnetic field dependence (nuclear magnetic relaxation dispersion, or NMRD profile) of T1 of exterior solvent water protons. Such data yield τ the mean lifetime of solvent molecules inside the vesicles, from τ = (fTlpara) – T1ves, where f is the volume fraction of entrapped water, T1Para is the observed T1 corrected for buffer background, and T1Ves is the relaxation time of water protons in the entrapped solution. For small spherical unilamellar vesicles of inner radius R, Pd = R/3τ. f can be obtained accurately from knowledge of both the concentration of Gd(DTPA)2− in the solution in which the vesicles were formed and the average concentration of ions in the final sample. At low temperatures, in the limit of slow exchange, T1Para becomes independent of field and τ = f T1Para; the observation of a field independent profile is a control that confirms that no paramagnetic material is external to the vesicles. We have measured T1Para, using a field‐cycling relaxometer, for suspensions of POPC (1‐palmitoyl‐2‐oleoyl‐lecithin) vesicles with 100–500 mM entrapped Gd(DTPA)2− and membrane concentrations of cholesterol ranging from 0 to 40 mol %. These profiles, which span the field range 0.01–50 MHz proton Larmor frequency, were taken at 5, 15, 25, and 35°C. Concentrations of Gd(DTPA)2− were determined independently by both ICP analyses and NMRD methods. Values for Pd for vesicles with 100 mM Gd(DTPA)2− and outer diameters 100 nm ± 20%, as determined by quasielastic light scattering, are 63, 47, 24, 16, and 8.7 × 10−4cm s−1, at 25°C, for cholesterol concentrations of 0, 10, 20, 30, and 40%, respectively. The corresponding activation enthalpies are 14, 14, 14, 17, and 17 kcal/M. Comparison with 2H NMR studies of deuterated POPC vesicles with no cholesterol at 20°C, and with 10% at 40°C, which yielded the same order parameter for the palmitoyl acyl chains, gives no indication of a correlation between order parameter and permeability.