Partial structural analysis of concanavalin A binding glycopeptides of large size from human teratocarcinoma-derived cells: cleavage by hydrazinolysis and alkaline borohydride.
Pronase digests of cultured teratocarcinoma-derived cells (PA 1) of human origin have been previously shown to contain large-sized glycopeptides (relative mass (Mr) greater then 7400), of which 15-23% are retained by columns of concanavalin A (Con A) - Sepharose and can be eluted with 10 mM methyl alpha-D-mannopyranoside. The present data show that this fraction (A - Con A II) contains a family of glycopeptides that are degradable with anhydrous hydrazine as well as with 0.05 M NaOH - 1 M NaBH4. The cleavage products representing individual oligosaccharide chains, presumably as oligosaccharides and glycopeptides, consisted mostly of medium- (Mr 1400-6000) and small-sized (Mr less than 1400) molecules. This implies that glycopeptides bearing several oligosaccharide chains were present in A - Con A II. Most of the individual oligosaccharide chains were not bound to Con A - Sepharose, but some were retained by the lectin column in the same way as the original glycopeptides. Some of the oligosaccharides were degraded partially with endo-beta-galactosidase from Escherichia freundii suggesting the presence of GalbetaGlcNAcbeta repeats. The present findings show that A - Con A II may be different from the "embryonic" glycopeptides of mouse teratocarcinoma cells that are reportedly not cleaved by mild alkaline borohydride treatment. Instead, A - Con A II is reminiscent of the T-1 glycopeptide of glycophorin.
D001894:Borohydrides; D002460:Cell Line; D055598:Chemical Phenomena; D002621:Chemistry; D005260:Female; D006801:Humans; D006834:Hydrazines; D006863:Hydrogen-Ion Concentration; D009844:Oligosaccharides; D010051:Ovarian Neoplasms / Q000032:analysis*; D011952:Receptors, Concanavalin A / Q000302:isolation & purification; D013724:Teratoma / Q000032:analysis*
D001894:Borohydrides; D006834:Hydrazines; D009844:Oligosaccharides; D011952:Receptors, Concanavalin A; C027774:concanavalin A-binding glycoproteins; C029424:hydrazine; C025364:sodium borohydride