Analysis of Glycosphingolipids by Fluorophore-Assisted Carbohydrate Electrophoresis Using Ceramide Glycanase from Mercenaria mercenaria
✍ Scribed by S.S. Basu; S. Dastgheibhosseini; G. Hoover; Z.X. Li; S. Basu
- Publisher
- Elsevier Science
- Year
- 1994
- Tongue
- English
- Weight
- 509 KB
- Volume
- 222
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
Polyacrylamide gel electrophoresis of fluorophorelabeled saccharides is a simple and sensitive separation method for glycan analysis. 'This method requires an aldehydic reducing carbon on the saccharide in order to react with the amine group of the fluorophore ((8-\mathrm{ami}) nonaphthalene-1,3,6-trisulfonic acid or 2 -aminoacridone). We have used exoglycosidase-free ceramide glycanase from hard-shelled clam to expose the reducing terminal of the glycan in glycosphingolipid by cleaving the linkage between the glycan and the ceramide. The released glycan was used for fluorophore-assisted carbohydrate electrophoresis, in order to qualitatively determine its monosaccharide composition, chain length, and glycosidic linkages using linkage-specific glycosidases, including a (\beta 1-3,4) galactosidase from clam. c. 1994 Academic Press, Inc.