Journal article
Large-scale preparation and characterization of N-linked glycopeptides from bovine fetuin
Analytical biochemistry, Vol.184(2), pp.249-258
1990
DOI: 10.1016/0003-2697(90)90676-Z
PMID: 1691598
Abstract
A preparative scheme has been developed to purify asialo-glycopeptides from each of the three N-linkage sites of bovine fetuin, allowing the isolation of 100-μmol quantities of asialo-glycopeptides from 20 g of fetuin. The procedure yields seven asialo-glycopeptides which were determined to be 95% homogenous in peptide and oligosaccharide structure. The isolation scheme uses two high-capacity reverse-phase eluant systems. The primary RP-HPLC purification performed with boric acid buffered to pH 7 with triethylamine resolved sialylated tryptic glycopeptides simultaneously on the basis of glycosylation site and degree of sialylation. A second RP-HPLC purification was performed eluting isocratically with dilute phosphoric acid which resolved residual peptide and oligosaccharide heterogeneity from asialo-glycopeptides containing short peptides. Structural characterization of the products was performed utilizing 400-MHz proton NMR spectroscopy and amino acid and monosaccharide analysis. The glycopeptides contain two previously identified variant triantennary oligosaccharides which possess either Galβ(1 → 4) or Galβ(1 → 3) linkages to
N-acetylglucosamine at one terminal branch or a biantennary oligosaccharide. These compounds should prove to be invaluable in studying carbohydrate-protein interactions, such as binding by the
Gal
GalNAc
lectin of mammalian hepatocytes, in the detailed three-dimensional structural analysis of complex oligosaccharides, and as purified substrates for the study of the action of glycoconjugate-modifying enzymes.
Details
- Title: Subtitle
- Large-scale preparation and characterization of N-linked glycopeptides from bovine fetuin
- Creators
- Kevin G RiceNarasinga B.N RaoYuan C Lee
- Resource Type
- Journal article
- Publication Details
- Analytical biochemistry, Vol.184(2), pp.249-258
- DOI
- 10.1016/0003-2697(90)90676-Z
- PMID
- 1691598
- NLM abbreviation
- Anal Biochem
- ISSN
- 0003-2697
- eISSN
- 1096-0309
- Publisher
- Elsevier Inc
- Language
- English
- Date published
- 1990
- Academic Unit
- Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984065319502771
Metrics
15 Record Views