HPLC method for the determination of Fuc to Asn-linked GlcNAc fucosyltransferases
HPLC method for the determination of Fuc to Asn-linked GlcNAc fucosyltransferases
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HPLC 法测定 Fuc 至 Asn 连接的 GlcNAc 岩藻糖基转移酶
DOI：
10.1023/a:1006951802623
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发表时间：
2004
影响因子：
3
通讯作者：
F. Altmann
中科院分区：
文献类型：
--
作者：
Andreas Roitinger;H. Leiter;E. Staudacher;F. Altmann
哺乳动物细胞通常含有一种酶，可将岩藻糖转移到具有α 1，6-键的N-聚糖的还原末端GlcNAc（GlcNAc-1）上。另一方面，在植物中，岩藻糖通过α 1，3-键转移到GlcNAc-1。昆虫细胞可以表现出两种酶活性。目前，这些岩藻糖基转移酶的活性已经通过将放射性标记的岩藻糖掺入受体糖肽中来确定。然而，该测定不能区分这两种活性。在这里，我们报告使用丹磺酰化糖天冬酰胺的具体测定1，3-和1，6-岩藻糖基转移酶。两种可能的产物和底物在反相柱上分离并通过荧光检测。
Mammalian cells often contain an enzyme which transfers fucose onto the reducing terminal GlcNAc (GlcNAc-1) of N-glycans with an α1,6-linkage. In plants, on the other hand, the fucose is transferred to GlcNAc-1 with an α1,3-linkage. Insect cells can exhibit both enzymatic activities. Hitherto, the activity of these fucosyltransferases has been determined by the incorporation of radioactively labelled fucose into an acceptor glycopeptide. This assay, however, cannot discriminate these two activities. Here we report on the use of dansylated glycoasparagine for the specific determination of 1,3- and 1,6-fucosyltransferases. The two possible products and the substrate are separated on a reversed phase column and detected by fluorescence.