Mannose containing glycopeptides of cells derived from human teratocarcinoma (line PA 1)

Abstract

Human teratocarcinoma-derived cells, line PA 1, maintained in the undifferentiated state, yielded upon exhaustive pronase digestion unusually large glycopeptides (fraction A), which showed on gel filtration an apparent MW > 7400. These glycopeptides derived from whole cell proteins carried large-sized oligosaccharides as evidenced by repeated pronase treatments, hydrazinolysis and .beta.-elimination experiments. The oligosaccharides consisted of mannose, fucose, galactose and N-acetylglucosamine. The PA 1 cells contained also oligomannosyl type glycans, presumably linked to asparagine (fraction C glycopeptides). These glycopeptides were strongly bound to Con [concanavallin] A-sepharose and their oligosaccharides were released by endo-.beta.-N-acetylglucosaminidase H. The liberated glycans ranged from Man5GlcNAc to Man9 GlcNAc as analyzed by paper chromatography. Pulse-chase experiments suggest that there is a precursor-product relationship between the mannose label in the fraction C (oligomannosyl type) glycopeptides and the fraction A glycopeptides.