By Elizabeth F. Hounsell
Glycoprotein research in Biomedicine brings jointly a complete diversity of protocols regarding carbohydrate biochemistry. the 1st 1/2 the guide outlines physiochemical (MS and NMR), chemical, and chromatographic ideas for outlining the buildings and variety of oligosaccharide sequences. the second one part describes extra organic and immunological methods used to notice alterations in glycosylation styles in disorder.
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Additional resources for Glycoprotein Analysis in Biomedicine (Methods in Molecular Biology)
Release and Isolation of O-Linked Oligosaccharides from Glycoproteins 1. lM) containing 2MNaBH4: Add NaBH* (store dry) to a stock solution of KOH. Make a new solution every time. 2. OM). 3. Ion-exchange resin, AG50Wx8 (Bio-Rad, Richmond, CA) 100-200 mesh, hydrogen form: Wash in distilled water and methanol on a sintered funnel. Store in methanol, and wash with water before use. 4. 0 L; adjust pH with pyridine or acetic acid. 5. DEAE-Sephadex A-25 (Pharmacia, Uppsala, Sweden): Swell the gel for at least 3 h at room temperature.
Interpretation of Mass Spectra (see Note 8) 1. A guide for interpretating mass spectra of periodate degraded N-and Olinked glycoprotein oligosaccharides is shown in Tables 1 and 2. FAB-mass spectra of permethylated Nacetylhexosamine containing oligosaccharides are characterized by intense ions formed by cleavage of the 2-acetamido-2deoxyhexosyl linkages. Binding positions between monosaccharide residues are determined from the primary and secondary sequence ions. The monosaccharide sequence from the nonreducing terminal is determined from the primary ions formed after cleavage of the glycosidic bonds.
Chem. 254,9795-9799. 14. , and Osawa, T. (1981) Structure of a complex-type sugar chain of human glycophorin A. Bzochemestry 20,566-566. 15. , Goldstem, I. , Van Damme, E. J. , and Peumans, W. J. (1988) Binding properties of a mannose-specific lectin from the snowdrop (Galanthus nivales) bulbJ. Beol. Chem. 263,728-734. 16. , and Osawa, T. (1981) Structural requirements for the binding of ohgosaccharides and glycopeptides to immobihzed wheat germ agglutinin Biochemistry 20,5894-5899. 17. , and Kobata, A.
Glycoprotein Analysis in Biomedicine (Methods in Molecular Biology) by Elizabeth F. Hounsell