Calcium modulates the mechanical properties of anionic phospholipid membranes

Using micropipette aspiration and fluorescence techniques, we have studied the material properties of charged lipid vesicles in calcium solutions. Vesicles were composed of phosphatidylglycerol (PG)/phosphatidylcholine (PC) or phosphatidic acid (PA)/PC mixtures. For the case of PG/PC membranes, we measure no effect of anionic lipid fraction on elasticity but a monotonic decrease up to 20% for tension required to induce membrane failure. Both of these observations are rationalized by a model we have developed to describe membrane electrostatic interactions in a two-component salt solution and the resulting changes in membrane properties. Critical tensions measured for PA/PC membranes, on the other hand, did not depend on anionic lipid fraction and were uniformly approximately 35% lower than PG/PC vesicles. This is likely due to a lateral phase separation in the membrane. By combining mechanical properties with fluorescence observations we propose that the PA-rich phase separates into small unconnected domains.