Evaluation of Young's Modulus of Tethered 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine Membranes Using Atomic Force Spectroscopy

Unilamellar vesicles composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) with varying 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-poly(ethylene glycol)-2000-N-[3-(2-pyridyldithio) propionate] (DSPE-PEG-PDP) concentration between 0 mol % and 24 mol % were assembled on atomically flat template-stripped gold (TS Au) surfaces. Force spectroscopy, using an atomic force microscope (AFM), of the resulting tethered lipid bilayer membranes (tLBMs) in buffer provided information regarding mechanical response as a function of tethering molecule, DSPE-PEG-PDP, concentration. Youngs modulus was determined by fitting the forceindentation curve with a recently modified Sneddon model that corrects for contributions from the substrate underneath. At low concentrations, Youngs modulus is lower than that of a supported POPC LBM, i.e., directly sitting on a solid substrate. The decrease in modulus is attributed to increased membrane fluidity as coupling between the tLBM and solid substrate is reduced by the incorporation of DSPE-PEG-PDP tethering groups. From the determined Youngs modulus values, the PEG chain conformation is found to dominate tLBM rigidity at concentrations above 6 mol %. Analysis of AFM force spectroscopy data indicates that the poly(ethylene glycol) (PEG) mushroom to brush transition occurs near 6 mol %, and this leads to first softening and then abrupt stiffening of tLBMs at higher DSPE-PEG-PDP concentration associated with the transition. When DSPE-PEG-PDP concentration is increased to 24 mol %, AFM topography and Youngs modulus appear correlated with another phase transition; AFM topography images are consistent with a bilayer disk structure with DSPE-PEG-PDP segregated at the rim of the disk.