Fractionation of large glycopeptides of human teratocarcinoma-derived cells by concanavalin A-Sepharose chromatography.

Human teratocarcinoma derived cells, line PA 1, were labeled with radioactive monosaccharides and subsequently digested with pronase. Large sized glycopeptides (fraction A) were isolated by gel filtration on Bio-Gel P-10. Their chromatography on concanavalin A-Sepharose gave three subfractions, two of which were eluted with a sugar-free buffer and the third with 10 mM alpha-methyl mannoside. The first subfraction (fraction A-Con A Ia) incorporated label from [3H]galactose and [3H]glucosamine and contained the largest components of fraction A. The second and the third subfractions (fractions A-Con A Ib and A-Con A II) were glycopeptides which incorporated label from tritiated fucose, mannose, galactose, and glucosamine. Even these molecules were of large size eluting partially at the void volume from Bio-Gel P-60. The glycopeptides of fraction A-Con A Ib contained mannose, fucose, galactose, N-acetylglucosamine, and N-acetylgalactosamine. Fucose and galactose residues occupied ultimate or penultimate positions at the nonreducing termini of the oligosaccharides. N-Acetylneuraminic acid, too, was present in the glycopeptides of fraction A.