Structural analysis of oligosaccharide-alditols released by reductive beta-elimination from the jelly coats of the anuran Bufo arenarum

Abstract

Amphibian egg jelly coats are formed by components secreted along the oviduct. These secretion products overlay the oocytes as they are transported toward the cloaca. Mucin type glycoproteins are the major constituents of the egg jelly coats. In this study, the O-linked carbohydrate chains of the jelly coat surrounding the eggs of Bufo arenarum were released by alkaline borohydride treatment. Fractionation of the mixture of O-linked oligosaccharide-alditols was achieved by a combination of chromatographic techniques comprising gel-permeation chromatography, ion-exchange chromatography and high-performance liquid chromatography using an amino-bonded silica column, afforded 11 fractions. The primary structures of these O-glycans were determined by one-dimensional and two-dimensional 1H-NMR spectroscopy in conjunction with matrix-assisted laser-desorption-ionization--time-of-flight mass spectrometry. 11 oligosaccharide structures, possessing a core consisting of Galbeta1-->3GalNAc-ol with or without branching through a GlcNAc residue linked beta1-->6 to the GalNAc residue (core type 2 or core type 1, respectively) are described. These oligosaccharide-alditols with these types of cores have been identified previously in mammalian mucins or in oviducal amphibian jellies. These glycans contain blood group determinants such as H, A or Cad antigens.