Quantitative Analysis of Molecular Transport across Liposomal Bilayer by J-Mediated 13C Overhauser Dynamic Nuclear Polarization

We introduce a new NMR technique to dramatically enhance the solution-state 13C NMR sensitivity and contrast at 0.35 T and at room temperature by actively transferring the spin polarization from Overhauser dynamic nuclear polarization (ODNP)-enhanced 1H to 13C nuclei through scalar (J) coupling, a method that we term J-mediated 13C ODNP. We demonstrate the capability of this technique by quantifying the permeability of glycine across negatively charged liposomal bilayers composed of dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG). The permeability coefficient of glycine across this DPPC/DPPG bilayer is measured to be (1.8 ± 0.1) × 10–11m/s, in agreement with the literature value. We further observed that the presence of 20 mol % cholesterol within the DPPC/DPPG lipid membrane significantly retards the permeability of glycine by a factor of 4. These findings demonstrate that the high sensitivity and contrast of J-mediated 13C ODNP affords the measurement of the permeation k...