The biosynthesis of gangliosides. The incorporation of galactose, N-acetylgalactosamine and N-acetylneuraminic acid into endogenous acceptors of subcellular particles from rat brain in vitro

Abstract

Gangliosides bound to subcellular particles from rat brain were labelled by incubation of the particles (i) with CMP-N[3H]-acetylneuraminic acid and (ii) simultaneously, with CMP-N[3H]-acetylneuraminic acid and UDP-N-acetyl-[14C1]galactosamine or with CMP-N[3H]-acetylneuraminic acid and UDP-[U-14C]-galactose. Analysis of the labelled gangliosides showed that in (i), (a) the labelling was mostly in the neuraminidase-labile sialyl groups, (b) rigid relationships exist between the enzymes and the sialyl acceptors; the enzymes are not free to interact with all the specific substrates present in the preparation and (c) the precursor of the trisialoganglioside was the major disialoganglioside with a sialyl 2→8 sialyl group. In (ii), (a) precursor–product relationships between the main pools of each ganglioside apparently do not exist, (b) for the labelling of Tay–Sachs ganglioside the amount formed from hematoside was at least 2.5 times that from aminoglycolipid and (c) the major monosialoganglioside was the precursor for the major disialoganglioside with a sialyl 2→8 sialyl group.