1H NMR characterization of a hen ovalbumin tyrosinamide N-linked oligosaccharide library

M L Corradi Da Silva; Hilary J Stubbs; Toshiaki Tamura; Kevin G Rice

doi:10.1006/abbi.1995.1255

Back

1H NMR characterization of a hen ovalbumin tyrosinamide N-linked oligosaccharide library

Journal article

Peer reviewed

1H NMR characterization of a hen ovalbumin tyrosinamide N-linked oligosaccharide library

M L Corradi Da Silva, Hilary J Stubbs, Toshiaki Tamura and Kevin G Rice

Archives of biochemistry and biophysics, Vol.318(2), pp.465-475

04/20/1995

DOI: 10.1006/abbi.1995.1255

PMID: 7733678

View Online

Abstract

A library of 15 N-linked oligosaccharide structures was prepared from ovalbumin and characterized using high-field NMR and mass spectrometry. The oligosaccharides were enzymatically released from ovalbumin glycopeptides, and the reducing ends were reacted with ammonium bicarbonate to form oligosaccharide-glycosylamines. These reacted with Boc-tyrosine-N-hydroxysuccinimide ester, resulting in a mixture of tyrosinamide-oligosaccharides. The Boc group was removed to expose an amine terminus which enhanced the resolution of tyrosinamide-oligosaccharides when chromatographed on reverse-phase HPLC. Ten major and five minor oligosaccharides were purified on a micromole scale and characterized using 1H NMR and FAB-MS. The structures include high-mannose, hybrid, and complex oligosaccharides possessing from two to five antenna, providing the most complete definition of ovalbumin N-linked oligosaccharides to date. The resulting library is well suited to biological studies due to the presence of a single terminal tyrosine residue on each oligosaccharide that allows radioiodination or the attachment of additional probes to these glycoconjugates prior to biological studies.

Carbohydrate Sequence

Magnetic Resonance Spectroscopy - methods

Hydrogen

Glycopeptides - chemistry

Glycopeptides - isolation & purification

Molecular Sequence Data

Chromatography, High Pressure Liquid

Tyrosine - analogs & derivatives

Oligosaccharides - isolation & purification

Trypsin

Animals

Chickens

Oligosaccharides - chemistry

Spectrometry, Mass, Fast Atom Bombardment

Female

Carbohydrate Conformation

Ovalbumin - chemistry

Databases, Factual

Details

Title: Subtitle: 1H NMR characterization of a hen ovalbumin tyrosinamide N-linked oligosaccharide library
Creators: M L Corradi Da Silva - Division of Pharmaceutics and Pharmaceutical Chemistry, College of Pharmacy, Ohio State University, Columbus 43210, USA
Hilary J Stubbs
Toshiaki Tamura
Kevin G Rice
Resource Type: Journal article
Publication Details: Archives of biochemistry and biophysics, Vol.318(2), pp.465-475
DOI: 10.1006/abbi.1995.1255
PMID: 7733678
NLM abbreviation: Arch Biochem Biophys
ISSN: 0003-9861
eISSN: 1096-0384
Publisher: United States
Grant note: GM48048 / NIGMS NIH HHS
Language: English
Date published: 04/20/1995
Academic Unit: Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Medicinal and Natural Products Chemistry
Record Identifier: 9984065501202771

Metrics

17 Record Views

61 Times Cited - Web of Science