Structural characterization of the N-linked carbohydrate chains from mouse zona pellucida glycoproteins ZP2 and ZP3.
N-linked oligosaccharides of mouse zona pellucida glycoproteins ZP2 and ZP3 were prepared as pyridylaminated derivatives. Anion-exchange HPLC revealed that 95% or more of them are acidic. About 80% of these acidic chains were neutralized by digestion with Arthrobacter ureafaciens sialidase and a small amount of sulfate was found in the residual acidic chains. The neutralized fractions (SN fractions) from ZP2 and ZP3 were similarly fractionated by size-fractionation HPLC into many peaks having different numbers of LacNAc in the non-reducing regions. The SN fractions from ZP2 were cleaved by endo-beta-galactosidase into the core and non-reducing regions. The tri- and tetra-antennary complex-type chains with a Fuc residue were predominant in the core region. On the other hand, four kinds of fragment containing the LacNAc sequence were obtained from the non-reducing region. Sialic acids were shown to be linked to the core fragments, as well as the non-reducing fragments. The N-linked oligosaccharides of ZP3 were suggested to have essentially the same structures as those of ZP2.
Noguchi
S
S
Graduate School of Science and Technology, Chiba University, Japan.
Nakano
M
M
D000818:Animals; D002240:Carbohydrate Sequence; D005591:Chemical Fractionation; D004527:Egg Proteins; D005260:Female; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification; D051379:Mice; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis* / Q000302:isolation & purification; D010053:Ovary / Q000737:chemistry*; D011956:Receptors, Cell Surface; D015044:Zona Pellucida / Q000737:chemistry*; D000072216:Zona Pellucida Glycoproteins; D001616:beta-Galactosidase
D004527:Egg Proteins; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins; C000606520:Zp2 protein, mouse; C000606521:Zp3 protein, mouse; D001616:beta-Galactosidase