Electron-spin resonance study of aggregation of gramicidin in dipalmitoylphosphatidylcholine bilayers and hydrophobic mismatch.

The effect of aggregation of gramicidin A' (GA) on the phase structure of dipalmitoylphosphatidylcholine (DPPC) multilamellar vesicles was studied by cw-ESR using a chain-labeled lipid (16PC) at temperatures between 30 degrees and 45 degreesC that span the main phase transition of DPPC. Boundary lipids were observed only in dispersions with GA/DPPC molar ratios >1:15, where GA aggregates. Detailed fits by nonlinear least squares (NLLS) methods are consistent with the boundary lipid being characterized by a large negative order parameter ( approximately -0.4), indicative of a dynamic bending of the end of the acyl chain, and a substantially reduced motion, about an order of magnitude slower than that of the bulk lipid. The NLLS analysis compares favorably with a recent two-dimensional Fourier transform ESR study on DPPC/GA vesicles, which accurately discerned the bulk lipid. The detailed ESR observables are discussed in terms of the ordering effect of GA at low concentration of GA, the dissociation of the GA channel and the dynamic bending of the end chain segment of boundary lipid at high concentration of GA, and of HII phase formation induced by GA. It is suggested that these phenomena can be interpreted in terms of the combined effects of partial dehydration of the lipid headgroup by the GA and of the hydrophobic mismatch between GA and DPPC molecules. Substantial hysteresis is observed for heating versus cooling cycles, but only for a GA/DPPC molar ratio >1:15. This is consistent with the aggregation of GA molecules at high concentrations.