Gangliosides were isolated from bovine adrenal medulla and separated by thin-layer chromatography into five fractions. Total yield was 223 μg of lipid-bound sialic acid/g of fresh tissue, equivalent to about 840 μg of ganglioside. This amounted to 0.68 μmole of ganglioside, which is slightly over 50% the molar concentration in gray matter of brain. In terms of micromoles of ganglioside per gram of lipid, the adrenal medulla contained 75% as much as gray matter. Approximately 92% of the mixture consisted of two hematosides, AG5 and AG6, which contained N-glycolyl- neuraminic acid (NGNA) and N-acetylneuraminic acid (NANA), respectively. Both contained 1 unit each of glucose and galactose but no hexosamine. The remainder was a mixture of slow-migrating, brain-type gangliosides. One of these, AG3, was found by gasliquid partition analysis to contain glucose, galactose, galactosamine, and NANA in molar ratios of 1:1.93: 0.96:1.98. One of the NANA units was removed by neuraminidase. Fraction AG2 gave a similar though not identical analysis, except that approximately half the sialic acid was NGNA. It was not homogeneous, but the major component of AG2 interestingly contained both NANA and NGNA in the same structure. Fraction AG1 was a mixture but gave indication of another ganglioside containing both types of sialic acid.
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