Hydrolysis of di- and trisialo gangliosides in micellar and liposomal dispersion by bacterial neuraminidases

Abstract

The hydrolysis of di- and trisialo gangliosides by bacterial [Vibrio cholerae and Clostridium perfringens] neuraminidases was investigated. Slow rates of hydrolysis were obtained with micellar dispersions of the pure gangliosides; the rates increased considerably with mixtures of ganglioside and phospholipids, such as phosphatidylcholine or sphingomyelin. The greatest rates of hydrolysis were obtained with mixtures containing 5-10 mol % ganglioside and 90-95% phospholipid. With the aid of the nonpenetrating reagent trinitrobenzenesulfonic acid, it was ascertained that this mixture consisted of sealed, unilamellar vesciles in which the ganglioside was distributed symmetrically between the 2 layers of the liposome. When the relative proportion of the ganglioside was increased, the dispersions contained liposomes admixed with micelles of ganglioside and phospholipid. The rates of hydrolysis of the ganglioside could be correlated with the percentage of sealed vesicles in each mixture. Experiments in which another ganglioside (GM1) or cholesterol was incorporated into the mixed dispersions further supported this conclusion. The rate of hydrolysis is affected predominantly by interactions between the carbohydrate chains of ganglioside molecules. Ganglioside metabolism can be best studied when the latter are part of biological or model membranes.