Structural characterization of neutral oligosaccharides of human midcycle cervical mucin.
It was previously shown that alkaline borohydride treatment of human midcycle cervical mucin releases a heterogeneous population of reduced neutral, sialylated, and sulfated oligosaccharides (Yurewicz, E. C., and Moghissi, K. S. (1981) J. Biol. Chem. 256, 11895-11905). Three major neutral oligosaccharides were isolated with approximate compositions of Fuc:Gal:GlcNAc:N-acetylgalactosaminitol (GalNAcol) = 0:2:1:1 (A1), 1:2:1:1 (A2), and 2:2:1:1 (A3). They comprised roughly 21%, 13%, and 8% of human cervical mucin oligosaccharide chains, respectively. In the present report, each was analyzed by periodate oxidation, methylation, and sequential degradation with glycosidases. A1 was shown to contain more than one component, but structural analyses clearly demonstrated the presence of one predominant (75%) tetrasaccharide. The proposed structure, Gal beta 1-4GlcNAc beta 1-6(Gal beta 1-3)GalNAcol, has previously been found in human gastric, submaxillary, and ovarian cyst mucins in their carbohydrate-to-protein linkage regions. beta-Galactosidase from Aspergillus niger selectively cleaved the Gal beta 1-4GlcNAc linkage in the intact tetrasaccharide. Enzymatic hydrolysis of the Gal beta 1-3GalNAcol linkage required prior removal of the Gal beta 1-4GlcNAc beta 1-unit attached to 0-6 of GalNAcol. The data for A2 indicated a mixture of two oligosaccharides, Gal beta 1-4,3(Fuc alpha 1-3,4)GlcNAc beta 1-6(Gal beta 1-3)GalNacol and Fuc alpha 1-2Gal beta 1-4GlcNac beta 1-6(Gal beta 1-3)-GalNacol, in an approximate molar ratio of 3 to 4:1, respectively. Two structures are consistent with the data obtained for A3: Fuc alpha 1-2Gal beta 1-4,3(Fuc alpha 1-3,4)GlcNAc beta 1-6(Gal beta 1-3)GalNAcol and/or Gal beta 1-4,3(Fuc alpha 1-3,4)GlcNac beta 1-6(Fuc alpha 1-2Gal beta 1-3)GalNacol. The results indicate that A1 represents the "core" tetrasaccharide of the larger human cervical mucin oligosaccharides A2 and A3.
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002584:Cervix Uteri / Q000032:analysis*; D005260:Female; D006801:Humans; D008598:Menstruation; D008745:Methylation; D009077:Mucins / Q000302:isolation & purification*; D009092:Mucous Membrane / Q000032:analysis*; D009844:Oligosaccharides / Q000032:analysis*; D010084:Oxidation-Reduction; D010504:Periodic Acid
D009077:Mucins; D009844:Oligosaccharides; D010504:Periodic Acid