Specific radioactive labeling of terminal n-acetylgalactosamine of glycosphingolipids by the galactose oxidase-sodium borohydride method.
The galactose oxidase-sodium borohydride method was used to specifically label the terminal N-acetylgalactosamine of three glycosphingolipids, Gm2-ganglioside, asialo-Gm2-ganglioside, and globoside. All of the compounds showed a minimum of 95% radiopurity, and generally more than 90% of the total radioactivity was located in the terminal galactosamine moiety. Globoside and asialo-Gm2-ganglioside were labeled to high specific activities comparable with those of the sphingolipids with a terminal galactose moiety, labeled with the same procedure. These labeled compounds were well suited as substrates for the study of specific sphingolipid N-acetylgalactosaminidase. Gm2-ganglioside, however, was a poor substrate for galactose oxidase, and its specific activity was only a small percentage of the others. Furthermore, because of the low specific activity of the galactosamine moiety, it was necessary to pretreat Gm2-ganglioside with unlabeled sodium borohydride to reduce the nonspecific labeling of other portions of the molecule. The use of labeled sodium borohydride of a very high specific activity may yield specifically labeled Gm2-ganglioside suitable for metabolic studies. Thus, the method is useful for labeling not only terminal galactose but also terminal N-acetylgalactosamine of glycosphingolipids.
D000429:Alcohol Oxidoreductases; D001894:Borohydrides; D001923:Brain Chemistry; D005688:Galactosamine; D005732:Gangliosides; D006017:Glycolipids; D006801:Humans; D007553:Isotope Labeling; D008722:Methods; D012964:Sodium; D013107:Sphingolipids; D014316:Tritium
D001894:Borohydrides; D005732:Gangliosides; D006017:Glycolipids; D013107:Sphingolipids; D014316:Tritium; D005688:Galactosamine; D012964:Sodium; D000429:Alcohol Oxidoreductases