The interaction of calcium with gangliosides in bilayer membranes

We studied the binding of calcium to bilayer membranes formed from mixtures of phosphatidylcholine and mono-, di-, or trisialoganglioside by measuring its effect on the electrophoretic mobility of multilamellar vesicles and the conductance of planar bilayers. In 0.001 M monovalent salt solutions the surface potential of the membranes is large and micromolar concentrations of calcium have a significant effect on the mobility and conductance. In 0.1 M monovalent salt solutions the surface potential is small and millimolar concentrations of calcium are required to affect these parameters. The strong apparent binding of calcium we observed at low ionic strength could be due to the nonspecific accumulation of calcium in the electrical diffuse double layer. To distinguish between this nonspecific effect and binding of calcium to the membrane, we substituted dimethonium for calcium. Dimethonium is a divalent cation that screens negative charges but does not bind to lipids. We also examined the effect of replacing phosphatidylcholine by monoolein: calcium binds to phosphatidylcholine but not to monoolein. We describe our electrophoretic mobility results by combining the Poisson-Boltzmann and Navier-Stokes equations with the Langmuir adsorption isotherm. We conclude that calcium binds weakly to gangliosides with an intrinsic association constant of less than 100 M-1, which is similar to the association constant of calcium with phospholipids.