Isolation and characterization of alkali-labile oligosaccharide units from porcine erythrocyte glycophorin.

The oligosaccharide units of glycophorin isolated from porcine erythrocyte membranes were released by alkaline borohydride treatment and purified by gel filtration and ion-exchange chromatography. Structures of the O-glycosidic oligosaccharides were determined by methylation analysis, the methylated sugar being identified by gas-liquid chromatography-mass spectrometry and nitrous acid deamination after hydrazinolysis. The major oligosaccharide was a trisaccharide, Gal(1 leads to 3)[Neu-NGly(2 leads to 6)]GalNAc. The other oligosaccharides were larger and contained Glc-NAc. One was a pentasaccharide, Gal(1 leads to 3)Gal(1 leads to 4)GlcNAc(1 leads to 3)Gal(1 leads to 3)GalNAc. The structure of the trisaccharide was also analyzed by direct-probe mass spectrometry of the permethylated derivative, and the result obtained was consistent with the proposed structure.