Cholera toxin interactions with lipid bilayers

Abstract

The purpose of the experiments described in this paper was to assess the binding of cholera toxin to bilayers containing its receptor, the monosialoganglioside, GMl. The assay was based on the fact that GMl confers on the bilayer a negative surface charge. The magnitude of this surface charge was estimated by measuring the electrical conductance (G) of the bilayers exposed to nonactin-K+ under conditions where G is directly proportional to the potassium concentration in the aqueous solutions immediatey adjacent to the membrane surface. When bilayers were formed from mixtures of GMl and glycerolmonooleate (GMO), it was found that the molar ratio of the lipids in the bilayer was the same as that in the membrane forming solution. It was further found that cholera toxin or the binding subunit of the toxin (choleragenoid) bind to GMO bilayers containing GMl (but not to GMO bilayers containing phosphatidyl serine or disialoganglioside GDla). The value of the apparent dissociation constant for the binding of choleragen to its receptor was found to be 10(-11) M, comparable to values found in intact cells.