Stack R J, Sullivan M T
Glycomed, Incorporated, Alameda, CA 94501.
Glycobiology. 1992 Feb;2(1):85-92. doi: 10.1093/glycob/2.1.85.
The relative mobilities of various N-linked oligosaccharides reductively aminated to the charged fluorophore 8-amino-naphthalene-1,3,6-trisulphonic acid (ANTS) were determined by electrophoresis on high-density polyacrylamide slab gels. Each ANTS-derivatized oligosaccharide was assigned a relative migration index (RMI) expressed in terms of glucose equivalents, which was conveniently estimated by reference to a homologous series of ANTS--maltooligosaccharides run on each gel as oligosaccharide size standards. High-mannose-, complex- and hybrid-type structures were generally well resolved and easily visualized at picomole levels by simple UV light excitation. Application of these methods for the qualitative analysis of the oligosaccharides released from bovine fetuin and bovine asialofetuin by peptide-N-glycosidase F illustrates the usefulness of these techniques as fast, simple, and inexpensive tools for the characterization of N-linked oligosaccharides attached to glycoproteins.
通过在高密度聚丙烯酰胺平板凝胶上进行电泳，测定了还原胺化到带电荷荧光团8-氨基萘-1,3,6-三磺酸（ANTS）上的各种N-连接寡糖的相对迁移率。每个ANTS衍生化的寡糖都被赋予一个以葡萄糖当量表示的相对迁移指数（RMI），通过参考在每个凝胶上作为寡糖大小标准运行的一系列ANTS-麦芽寡糖同源物，可以方便地估算出该指数。高甘露糖型、复合型和杂合型结构通常能得到很好的分离，并且通过简单的紫外光激发，在皮摩尔水平就能轻松观察到。将这些方法应用于肽-N-糖苷酶F从牛胎球蛋白和牛去唾液酸胎球蛋白释放的寡糖的定性分析，说明了这些技术作为快速、简单且廉价的工具用于表征糖蛋白上连接的N-连接寡糖的实用性。
