Isolation of a novel O-linked, sulfated polysaccharide of high molecular weight from an ovarian cyst glycoprotein having blood group "A" activity.
Treatment of a blood group A-active ovarian cyst mucin glycoprotein with alkaline borohydride under conditions expected to cleave O-glycosidic linkages between carbohydrate and peptide releases a sulfated polysaccharide of average molecular weight 20,000. Its peptide and mannose content is less than 1%, and carbohydrate analysis gives Fuc/GalNAc/Gal/GlcNAc in the ratio of 1:1:2.2:2.2. Galactosaminitol is recovered at the level of one residue per 112-residue average polysaccharide chain. The 13C- and 1H-NMR spectra show that the polysaccharide has side chains whose non-reducing terminals have the blood group A structure on a type 1 chain: (Formula: see text). Methylation analysis confirms the presence of these blood group A type 1 sidechains as well as 4-substituted GlcNAc, 3-substituted galactose and 3,6-substituted galactose branch points. Periodate oxidation removes all the fucose and GalNAc from the non-reducing terminal but leaves intact the backbone composed of beta-linked Gal and GlcNAc, as would be expected for a polylactosamine. Although the native polysaccharide is resistant to endo-beta-galactosidase digestion, the product of periodate degradation is partially digested, giving a 30% yield of a trisaccharide shown by 1H-NMR spectroscopy to be: Gal(beta 1----3)GlcNAc(beta 1----3)Gal We conclude that this is a high molecular weight sulfated polysaccharide which is related to the asparagine-linked polylactosamine chains of cell surface glycoproteins which have been implicated in cell differentiation. However, the blood group A polysaccharide from the ovarian cyst mucin is unique in several respects. It is linked to the protein by an O-glycosidic bond rather than the N-asparagine linkage of the previously known polylactosamines which have a trimannosyl core, and its blood group A side chains are on a type 1 core rather than type 2 which is found on other polylactosamines.
D000017:ABO Blood-Group System; D002241:Carbohydrates / Q000032:analysis; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry; D006023:Glycoproteins; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008745:Methylation; D008970:Molecular Weight; D010048:Ovarian Cysts / Q000378:metabolism*; D011134:Polysaccharides / Q000302:isolation & purification*; D013431:Sulfates / Q000302:isolation & purification
D000017:ABO Blood-Group System; D002241:Carbohydrates; D006023:Glycoproteins; D011134:Polysaccharides; D013431:Sulfates