Ihara Hideyuki, Tsukamoto Hiroki, Taniguchi Naoyuki, Ikeda Yoshitaka
Division of Molecular Cell Biology, Department of Biomolecular Sciences, Faculty of Medicine, Saga University, Saga, Japan.
Methods Mol Biol. 2013;1022:335-48. doi: 10.1007/978-1-62703-465-4_25.
N-Glycans with an α-fucose unit linked to the 6-position of the innermost GlcNAc are widely distributed among the animal kingdom, from worms and insects to human. This α1,6-linked fucosyl residue, frequently referred to as a core fucose, is formed via the action of an α1,6-fucosyltransferase, the mammalian ortholog which is systematically called FUT8. In mammals, it is well known that the extent of core-fucosylation in cellular and secreted glycoproteins varies, e.g., according to differentiation and carcinogenesis of the cells. This chapter describes a method for the sensitive and quantitative assay of FUT8 activity using a fluorescence-labeled oligosaccharyl asparagine derivative as the glycosyl acceptor substrate.
带有与最内层N-乙酰葡糖胺6位相连的α-岩藻糖单元的N-聚糖广泛分布于动物界，从蠕虫、昆虫到人类。这种α1,6-连接的岩藻糖基残基，常被称为核心岩藻糖，是通过α1,6-岩藻糖基转移酶的作用形成的，该酶在哺乳动物中的直系同源物被系统命名为FUT8。在哺乳动物中，众所周知，细胞和分泌型糖蛋白的核心岩藻糖基化程度会有所不同，例如，根据细胞的分化和癌变情况。本章描述了一种使用荧光标记的寡糖基天冬酰胺衍生物作为糖基受体底物来灵敏且定量测定FUT8活性的方法。
