Kinetics of Calcium-Induced Fusion of Cell-Size Liposomes with Monolayers in Solutions of Different Osmolarity

Abstract

The effects of osmolarity, calcium concentration and cell-size liposomes in the subphase on the surface tension of phospholipid monolayers were investigated. The monolayers were spread from chloroform solutions of phosphatidic acid at air/water solution interface. The liposomes (of average diameter 3 μm) were formed from phosphatidic acid/egg lecithin (1:2) mixtures in water or 0.1 ᴍ water solutions of sucrose. For this system there were critical concentrations of calcium ions to produce a large reduction of the monolayer surface tension. The threshold calcium concentrations depended upon the sucrose concentration in the subphase. Without sucrose the threshold calcium concentration was 8 mᴍ, while for isoosmotic sucrose solutions (0.1/0.1 ᴍ in/out of liposome) it was 14 mᴍ. It sharply increased to 28 mᴍ CaCl2 at sucrose concentration difference across the liposome membrane 0.02 ᴍ and decreased to 26 mᴍ, 19 mᴍ, and 18 mᴍ with further increase of that difference to 0.04 ᴍ, 0.06 ᴍ, and 0.08 ᴍ, respectively. The rate of monolayer surface tension decrease was measured as a function of time at 30 mᴍ CaCl2 and different sucrose concentrations in the subphase solution. The initial rates at first decreased with increasing the osmotic pressure and after that they increased. The minimum occurred at sucrose concentration gradient across the liposome membrane 0.02 ᴍ, i.e., at the point of maximum threshold calcium concentration required for large decrease of the monolayer surface tension. These facts may be explained by recent theories of dynamics of adhesion, instability and fusion of membranes modeled as thin films.