TY - JOUR
T1 - Identification of the sialosyl-sialosyl linkage in biosynthesized gangliosides
AU - Yohe, Herbert C.
AU - Yu, Robert K.
N1 - Funding Information:
The authors thank Dr. Susumu Ando, Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan, for the g.l.c.-m-s. analysis of the sialic acid standards_ This work was supported by a grant (NS-11853) from the National Institutes of Health, and, in part, by grants from the National Multiple Sclerosis Society (RG 1289-A-2), the Esther A. and Joseph Klingestein Fund, and the C. G. Swebilius Fund.
PY - 1981/6/16
Y1 - 1981/6/16
N2 - A technique was developed to examine the sialosyl-sialosyl linkage in the minute quantities of complex gangliosides biosynthesized in vitro. The disialoganglioside, GD31 1 The ganglioside nomenclature used here is that described by Svennerholm. The gangliosides are designated as follows: GM3: II3NeuAc-LacCer; GM1: II3NeuAc-GgOse4Cer; GD3: II3(NeuAc)2LacCer; GD1a: IV3NeuAc, II3NeuAc-GgOse4Cer; and GT1a: IV3(NeuAc)2, II3NeuAc-GgOse4Cer., and the trisialoganglioside, GT1a, were biosynthesized as secondary products by the reaction of CMP-(4-14C)sialic acid with the glycolipid substrates, lactosyl ceramide and GM1 ganglioside, respectively. Following periodate oxidation-borohydride reduction, the intact sialic acid residue, and the susceptible sialic acid residue converted into an N-acetylheptulosaminic acid residue, were released by hydrolysis, esterified, separated, and analyzed for radioactivity. Analysis of the biosynthesized GD3 and GT1a gangliosides indicated radioactivity in both the susceptible and the nonsusceptible sialic acid residues, demonstrating that a sialosyl-(2→8)-sialosyl linkage had been synthesized. When the primary products (GM3 and GD1a, respectively) of the synthesis just described were examined, radioactivity was detected, as expected, only in the susceptible sialic acid residue. Because the N-acetylneuraminic acid and N-acetylheptulosaminic acid derivatives formed in the reaction were identified by chromatography, the standards used for comparison were verified by g.l.c.-m.s.
AB - A technique was developed to examine the sialosyl-sialosyl linkage in the minute quantities of complex gangliosides biosynthesized in vitro. The disialoganglioside, GD31 1 The ganglioside nomenclature used here is that described by Svennerholm. The gangliosides are designated as follows: GM3: II3NeuAc-LacCer; GM1: II3NeuAc-GgOse4Cer; GD3: II3(NeuAc)2LacCer; GD1a: IV3NeuAc, II3NeuAc-GgOse4Cer; and GT1a: IV3(NeuAc)2, II3NeuAc-GgOse4Cer., and the trisialoganglioside, GT1a, were biosynthesized as secondary products by the reaction of CMP-(4-14C)sialic acid with the glycolipid substrates, lactosyl ceramide and GM1 ganglioside, respectively. Following periodate oxidation-borohydride reduction, the intact sialic acid residue, and the susceptible sialic acid residue converted into an N-acetylheptulosaminic acid residue, were released by hydrolysis, esterified, separated, and analyzed for radioactivity. Analysis of the biosynthesized GD3 and GT1a gangliosides indicated radioactivity in both the susceptible and the nonsusceptible sialic acid residues, demonstrating that a sialosyl-(2→8)-sialosyl linkage had been synthesized. When the primary products (GM3 and GD1a, respectively) of the synthesis just described were examined, radioactivity was detected, as expected, only in the susceptible sialic acid residue. Because the N-acetylneuraminic acid and N-acetylheptulosaminic acid derivatives formed in the reaction were identified by chromatography, the standards used for comparison were verified by g.l.c.-m.s.
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U2 - 10.1016/S0008-6215(00)80747-8
DO - 10.1016/S0008-6215(00)80747-8
M3 - Article
AN - SCOPUS:2642699893
SN - 0008-6215
VL - 93
SP - 1
EP - 9
JO - Carbohydrate Research
JF - Carbohydrate Research
IS - 1
ER -