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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >SEPARATION OF 1-AMINOPYRENE-3,6,8-TRISULFONATE-LABELED ASPARAGINE-LINKED FETUIN GLYCANS BY CAPILLARY GEL ELECTROPHORESIS
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SEPARATION OF 1-AMINOPYRENE-3,6,8-TRISULFONATE-LABELED ASPARAGINE-LINKED FETUIN GLYCANS BY CAPILLARY GEL ELECTROPHORESIS

机译：毛细管凝胶电泳分离1-氨基A啶-3,6,8-三磺酸酯连接的天冬酰胺连接的牛磺酸甘油

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Asparagine-linked glycans of bovine fetuin were separated by capillary gel electrophoresis after enzymatic release (peptide-N-glycosidase F) and labeling via reductive amination by a fluorescent dye, 1-aminopyrene-3,6-8-trisulfonate (APTS). At low separation pH (2.5) only two dominant peaks were observed. Increasing the separation buffer pH to 4.75 resulted in complete separation of two primary doublets and several minor peaks from the fetuin N-linked glycan pool. Two of the four major peaks were spiked with purified individual standards and were identified as trisialylated triantennary structures with different sialylation linkages. The other two larger peaks were postulated to be tetrasialylated triantennary structures, based on calculations considering their corresponding glucose unit (GU) values. Effects of the electrophoretic separation parameters, such as gel concentration, electric field strength and temperature on the migration behavior of the two major doublets of the fetuin glycan pool were also thoroughly examined. Our data suggest that the capillary gel electrophoresis separation of the multisialylated branched oligosaccharides with different linkage isomers, released from bovine fetuin, is fundamentally based on their degree of sialylation and hydrodynamic volumes. [References: 30]
机译：酶促释放（肽-N-糖苷酶F）后，通过毛细管凝胶电泳分离牛胎蛋白的天冬酰胺连接的聚糖，并通过荧光染料1-氨基py-3,6-8-三磺酸盐（APTS）的还原胺化进行标记。在低分离pH（2.5）下，仅观察到两个主要峰。将分离缓冲液的pH值提高到4.75，可从胎球蛋白N-连接的聚糖池中完全分离出两个主要的双峰和几个次要峰。四个主要峰中的两个峰被纯化的单个标准物加标，并被鉴定为具有不同唾液酸化键的三唾液酸化三触角结构。根据考虑到它们相应的葡萄糖单位（GU）值的计算，假定其他两个较大的峰为四唾液酸化的三触角结构。还彻底检查了电泳分离参数（如凝胶浓度，电场强度和温度）对胎球蛋白聚糖池的两个主要双峰的迁移行为的影响。我们的数据表明，从牛胎球蛋白释放的具有不同键合异构体的多唾液酸化分支寡糖的毛细管电泳分离基本上是基于它们的唾液酸化程度和流体动力学体积。 [参考：30]

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