Abstract
Gangliosides GM2, GM1 and GD1b were radiolabelled at C-6 of the terminal galactose orN-acetylgalactosamine by the galactose oxidase/[3H]NaBH4 method; gangliosides GM2, GM1, Fuc-GM1 and GD1a were radiolabelled at C-3 of the long chain base by the 2,3-dichloro-5,6-dicyanobenzoquinone/[3H]NaBH4 method.
By application of an original HPLC procedure, eight different molecular species were prepared from each labelled ganglioside. Each of these species was characterized by the presence of one of the following long chain bases:erythro C18 sphingosine,threo C18 sphingosine,erythro C18 sphinganine,threo C18 sphinganine,erythro C20 sphingosine,threo C20 sphingosine,erythro C20 sphinganine andthreo C20 sphinganine.
From GD1b only the species containing theerythro forms of long chain bases were obtained.
The individual molecular species were more than 99% homogeneous and had a radiopurity better than 99%. The molecular species of the same ganglioside, radiolabelled at C-3 of the long chain base, had identical specific radioactivity, namely 1.17, 1.25, 0.85 and 1.28 Ci/mmol for GM2, GM1, Fuc-GM1 and GD1a respectively. The molecular species of the same ganglioside, radiolabelled at C-6 of terminal galactose orN-acetylgalactosamine, had similar specific radioactivity, namely 1.34–1.40, 1.44–1.51, 1.37–1.44 Ci/mmol for GM2, GM1 and GD1b respectively.
Similar content being viewed by others
References
Ledeen RW, Yu RK (1978) in Research Methods in Neurochemistry, eds Marks N, Rodnight R, Plenum, New York, p 371–410
Wiegandt H (1982) Adv Neurochem 4:149–223.
Svennerholm L, Fredman P, Elwing H, Holmgren J, Strannegard O (1980) ACS Symp Ser 128:373–90.
Fishman PH (1982) J Membr Biol 69:85–98.
Obata K, Olde M, Handa S (1977) Nature 266:359–71.
Morgan JI, Seifert W (1979) J Supramol Struct 10:111–24.
Roisen FG, Bartfeld H, Nagele R, Yorke G (1981) Science 214:577–78.
Leon A, Facci L, Toffano G, Sonnino S, Tettamanti G (1981) J Neurochem 37:350–57.
Facci L, Leon A, Toffano G, Sonnino S, Ghidoni R, Tettamanti G (1984) J Neurochem 42:299–305.
Ghidoni R, Sonnino S, Masserini M, Orlando P, Tettamanti G (1981) J Lipid Res 22:1286–95.
Suzuki K, Suzuki Y (1972) J Lipid Res 13:687–90.
Ghidoni R, Tettamanti G, Zambotti V (1974) Ital J Biochem 23:320–28.
Sonnino S, Ghidoni R, Gazzotti G, Kirschner G, Galli G, Tettamanti G (1984) J Lipid Res 25:620–29.
Svennerholm L (1977) Lipids 12:455–68.
Tettamanti G, Bonali F, Marchesini S, Zambotti V (1973) Biochim Biophys Acta 296:160–70.
Ghidoni R, Sonnino S, Tettamanti G, Baumann N, Reuter G, Schauer R (1980) J Biol Chem 255:6990–95.
Chigorno V, Sonnino S, Ghidoni R, Tettamanti G (1982) Neurochem Int 4:397–403.
Warren L (1959) J Biol Chem, 234:1971–75.
Svennerholm L (1957) Biochim Biophys Acta 24:604–11.
Novak A, Lowden JA, Gravel YL, Wolfe LS (1979) J Lipid Res 20:678–81.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gazzotti, G., Sonnino, S., Ghidoni, R. et al. Preparation of the tritiated molecular forms of gangliosides with homogeneous long chain base composition. Glycoconjugate J 1, 111–121 (1984). https://doi.org/10.1007/BF01213725
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01213725