Journal article
Tyrosine Derivatization and Preparative Purification of the Sialyl and Asialyl N-Linked Oligosaccharides from Porcine Fibrinogen
Archives of biochemistry and biophysics, Vol.312(1), pp.151-157
1994
DOI: 10.1006/abbi.1994.1293
PMID: 8031122
Abstract
The N-linked oligosaccharides from porcine fibrinogen were purified following their release from glycopeptides using N-glycosidase F. In separate experiments, both sialyl and asialyl oligosaccharides were prepared from 5 g of fibrinogen. The reducing oligosaccharides were reacted with ammonium bicarbonate to form oligosaccharide-glycosylamines and then derivatized with
tert-butoxycarbonyl-L-tyrosine
N-hydroxysuccinimidyl ester. Tyrosinamide-oligosaccharides were purified first by gel filtration chromatography and then by reverse-phase HPLC and the products were characterized by proton NMR and fast atom bombardment-MS. Porcine fibrinogen was found to have predominantly a single asialyl biantennary oligosaccharide containing a fucose linked α1-6 to GlcNAc 1. The oligosaccharide possesses two sialylation patterns with a major form (70%) having a single
N-acetyl neuraminic acid (NeuAc) residue linked α2-6 to galactose on only one antenna and a minor form (30%) possessing two NeuAc residues linked α2-6 to both terminal galactose residues. In addition to developing an isolation procedure and establishing the structures of porcine fibrinogen oligosaccharides, this study improves on the tyrosine derivatization technique as a general approach to isolate structurally diverse N-linked oligosaccharides from glycoproteins.
Details
- Title: Subtitle
- Tyrosine Derivatization and Preparative Purification of the Sialyl and Asialyl N-Linked Oligosaccharides from Porcine Fibrinogen
- Creators
- M.L.C DasilvaT TamuraT McbroomK.G Rice
- Resource Type
- Journal article
- Publication Details
- Archives of biochemistry and biophysics, Vol.312(1), pp.151-157
- DOI
- 10.1006/abbi.1994.1293
- PMID
- 8031122
- NLM abbreviation
- Arch Biochem Biophys
- ISSN
- 0003-9861
- eISSN
- 1096-0384
- Publisher
- Elsevier Inc
- Language
- English
- Date published
- 1994
- Academic Unit
- Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984065501702771
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