Structures of asparagine-linked oligosaccharides from hen egg-yolk antibody (IgY). Occurrence of unusual glucosylated oligo-mannose type oligosaccharides in a mature glycoprotein.
Asparagine-linked oligosaccharides present on hen egg-yolk immunoglobulin, termed IgY, were liberated from the protein by hydrazinolysis. After N-acetylation, the oligosaccharides were labelled with a UV-absorbing compound, p-aminobenzoic acid ethyl ester (ABEE). The ABEE-derivatized oligosaccharides were fractionated by anion exchange, normal phase and reversed phase HPLC, and their structures were determined by a combination of sugar composition analysis, methylation analysis, negative ion FAB-MS, 500 MHz 1H-NMR and sequential exoglycosidase digestions. IgY contained monoglucosylated oligomannose type oligosaccharides with structures of Glc alpha 1-3Man7-9-GlcNAc-GlcNAc, oligomannose type oligosaccharides with the size range of Man5-9GlcNAc-GlcNAc, and biantennary complex type oligosaccharides with core region structure of Man alpha 1-6(+/- GlcNAc beta 1-4)(Man alpha 1-3)Man beta 1-4GlcNAc beta 1-4(+/- Fuc alpha 1-6)GlcNAc. The glucosylated oligosaccharides, Glc1Man8GlcNAc2 and Glc1Man7GlcNAc2, have not previously been reported in mature glycoproteins from any source.
Ohta
M
M
Department of Biotechnology, Faculty of Engineering, Fukuyama University, Hiroshima, Japan.
Hamako
J
J
Yamamoto
S
S
Hatta
H
H
Kim
M
M
Yamamoto
T
T
Oka
S
S
Mizuochi
T
T
Matsuura
F
F
D010129:4-Aminobenzoic Acid; D000107:Acetylation; D000818:Animals; D001216:Asparagine / Q000378:metabolism*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006023:Glycoproteins / Q000737:chemistry*; D006031:Glycosylation; D007136:Immunoglobulins / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008358:Mannose / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism; D016339:Spectrometry, Mass, Fast Atom Bombardment
D006023:Glycoproteins; C016512:IgY; D007136:Immunoglobulins; D009844:Oligosaccharides; D001216:Asparagine; D008358:Mannose; D010129:4-Aminobenzoic Acid