Membrane Tension in Negatively Charged Lipid Bilayers in a Buffer under Osmotic Pressure.

Abstract

Osmotic pressure (Π) induces membrane tension in cell membranes and the lipid bilayers of vesicles and plays an important role in the functions and physical properties of these membranes. We recently developed a method to determine quantitatively the membrane tension of giant unilamellar vesicles (GUVs) under Π and applied it to GUVs comprising electrically neutral dioleoylphosphatidylcholine (DOPC). Here, we examined the effect of Π on GUVs composed of DOPC and negatively charged dioleoylphosphatidylglycerol (DOPG) in a buffer containing a physiological concentration of ions. First, we examined the rate constant, kr, for constant tension (σex)-induced rupture of DOPG/DOPC (4/6)-GUVs under Π and obtained the dependence of kr on σex in GUVs for various values of Π. Comparing this dependence in the absence of Π provided values for membrane tension due to Π, σosm, which agree with the theoretical values within the experimental error. The values of σosm for DOPG/DOPC-GUVs were smaller than those for DOPC-GUVs under the same Π. Two factors, that is, the solute concentration in a GUV suspension and the elastic modulus of the GUV membrane, can reasonably explain this difference based on the theory of σosm. We also examined the effect of Π on the rate constant, kFF, for the transbilayer movement of lipid molecules in single GUVs. The values of kFF increased with increasing Π, indicating that kFF increased with σosm. This result supports the existence of prepores in stretched lipid bilayers. Based on these results, we discuss the membrane tension of DOPG/DOPC-GUVs under Π.