Structural analysis of the N-linked carbohydrate chains of the 55-kDa glycoprotein family (PZP3) from porcine zona pellucida.
The N-linked oligosaccharides, released by hydrazinolysis from the major 55-kDa family, PZP3, of porcine zona pellucida glycoproteins, were separated into neutral (28%) and acidic (72%) carbohydrate chains by anion-exchange HPLC. By competition assay, it was shown that the mixture of neutral chains has the sperm-receptor activity, while that of the acidic chains has no activity. Their carbohydrate structures were analyzed after the reducing ends were modified with 2-aminopyridine. The neutral chains were fractionated into several components by reverse-phase and normal-phase HPLC. By sequential glycosidase digestion and 500-MHz 1H-NMR spectroscopy, the structures of three major components were determined. The structures of some of the minor components were analyzed only by sequential glycosidase digestion. By these analyses, it was found that a diantennary complex-type structure with a fucose residue was predominant in the neutral chains. Furthermore, the analyses of the endo-beta-galactosidase digests of the acidic chains revealed that the partially sulfated and sialylated N-acetyllactosamines are linked to the non-reducing ends of diantennary, triantennary, and tetra-antennary complex-type neutral chains, forming heterogeneous acidic chains.
Noguchi
S
S
Graduate School of Science and Technology, Chiba University, Japan.
Hatanaka
Y
Y
Tobita
T
T
Nakano
M
M
D000818:Animals; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D004527:Egg Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry; D011956:Receptors, Cell Surface; D013329:Structure-Activity Relationship; D013552:Swine; D015044:Zona Pellucida / Q000737:chemistry* / Q000276:immunology; D000072216:Zona Pellucida Glycoproteins
D004527:Egg Proteins; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins