We report a straightforward enzymatic synthesis of the 4-methylumbelliferyl glycoside of a complex-type oligosaccharide substrate for core fucosylation. We demonstrate the use of this synthetic glycoconjugate in a newly developed enzyme assay to probe the activity and inhibition of fucosyltransferase VIII, which catalyzes the core fucosylation of N-glycans on eukaryotic glycoproteins. In this fucosyltransferase assay, we use the fluorogenic probe and a specific glycosidase in a sequentially coupled enzyme reaction to distinguish an unmodified 4-methylumbelliferyl oligosaccharide probe from a fucosylated probe. Our findings show that this strategy is very sensitive and specific in its detection of enzyme activity and can even be used for analyzing impure tissue lysate samples.

中文翻译：

---

荧光报告基因底物的酶促合成和岩藻糖基转移酶VIII的高通量检测方法的发展提供了探测和抑制核心岩藻糖基化的工具包。

我们报告的复杂岩藻糖基化复杂类型寡糖底物的4-甲基伞形糖苷的直接酶法合成。我们证明了这种合成的糖缀合物在新开发的酶检测方法中的应用，以探测岩藻糖基转移酶VIII的活性和抑制，它催化了真核糖蛋白上N-聚糖的核心岩藻糖基化。在此岩藻糖基转移酶测定中，我们在顺序偶联的酶反应中使用了荧光探针和特异性糖苷酶，以将未修饰的4-甲基伞形寡糖寡糖探针与岩藻糖基化探针区分开。我们的发现表明，该策略在检测酶活性方面非常灵敏且特异，甚至可以用于分析不纯的组织裂解液样品。