Journal article
Reducing-end modification of N-linked oligosaccharides with tyrosine
Analytical biochemistry, Vol.216(2), pp.335-344
1994
DOI: 10.1006/abio.1994.1050
PMID: 7513972
Abstract
The N-linked oligosaccharides from bovine fetuin were purified using newly developed preparative purification methodology. N-linked oligosaccharides were released from tryptic glycopeptides utilizing N-glycosidase F on the 5-g scale. Selective desialylation with neuraminidase from Clostridium perfringens resulted in the formation of a mono-sialyl-oligosaccharide and asialo-oligosaccharides. The reducing ends of the oligosaccharides were converted to the glycosylamine and reacted with the N-hydroxysuccinimide ester of Boctyrosine. The tyrosinated oligosaccharides were resolved into individual peaks on RP-HPLC and then characterized by proton NMR and FAB-MS. A single asialo-triantennary, an asialo-biantennary, and a mono-sialyl-triantennary oligosaccharide were recovered in good yield. Each product contained a single Boc-Tyr residue attached to the reducing-end GlcNAc residue through a beta-glycosylamide linkage. The procedure was utilized to isolate multi-micromole quantities of oligosaccharides from gram quantities of glycoprotein, thus providing a new route to purify large quantities of N-linked oligosaccharides which contain a terminal tyrosine residue. The tyrosinated oligosaccharides are valuable glycoconjugate ligands which contain a chromophore that absorbs at 280 nm and has sufficient hydrophobicity to facilitate RP-HPLC separations. Furthermore, this group can be deprotected by removal of Boc to reveal a primary amine suitable for further derivatization and can also be radioiodinated for tracking during biological experiments.
Details
- Title: Subtitle
- Reducing-end modification of N-linked oligosaccharides with tyrosine
- Creators
- T TAMURA - Ohio State univ., coll. pharmacy, div. pharmaceutics pharmaceutical chemistry, Columbus OH 43210, United StatesM. S WADHWA - Ohio State univ., coll. pharmacy, div. pharmaceutics pharmaceutical chemistry, Columbus OH 43210, United StatesK. G RICE - Ohio State univ., coll. pharmacy, div. pharmaceutics pharmaceutical chemistry, Columbus OH 43210, United States
- Resource Type
- Journal article
- Publication Details
- Analytical biochemistry, Vol.216(2), pp.335-344
- DOI
- 10.1006/abio.1994.1050
- PMID
- 7513972
- NLM abbreviation
- Anal Biochem
- ISSN
- 0003-2697
- eISSN
- 1096-0309
- Publisher
- Elsevier; San Diego, CA
- Language
- English
- Date published
- 1994
- Academic Unit
- Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984065692002771
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