Analysis of glycosphingolipids by fluorophore-assisted carbohydrate electrophoresis using ceramide glycanase from Mercenaria mercenaria.
Analysis of glycosphingolipids by fluorophore-assisted carbohydrate electrophoresis using ceramide glycanase from Mercenaria mercenaria.
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DOI：
10.1006/abio.1994.1484
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发表时间：
1994-10
影响因子：
2.9
通讯作者：
S. Basu;S. Dastgheib-Hosseini;G. Hoover;Z. Li;S. Basu
中科院分区：
文献类型：
--
作者：
S. Basu;S. Dastgheib-Hosseini;G. Hoover;Z. Li;S. Basu
聚丙烯酰胺凝胶荧光标记电泳是一种简便、灵敏的聚糖分析方法。该方法需要糖上的还原碳，以便与荧光团的胺基（8-氨基萘-1，3，6-三磺酸或2-氨基吖啶酮）反应。我们已经使用来自硬壳蛤的不含外切聚糖酶的神经酰胺聚糖酶，通过切割聚糖和神经酰胺之间的连接来暴露鞘糖脂中聚糖的还原末端。将释放的聚糖用于荧光团辅助的碳水化合物电泳，以便使用连接特异性糖苷酶（包括来自蛤的β 1- 3，4半乳糖苷酶）定性确定其单糖组成、链长和糖苷键。
Polyacrylamide gel electrophoresis of fluorophore-labeled 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-aminonaphthalene-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.