Departamento de Química Orgánica, Facultad de Química, Universidad de Sevilla, Apartado 553, E-41012 Sevilla, Spain.
Bioorg Med Chem. 2010 Nov 1;18(21):7439-45. doi: 10.1016/j.bmc.2010.09.003. Epub 2010 Sep 7.
New fluorescently-labelled sp(2)-iminosugars based on the 5N,6S-[N'-(4-aminobutyl)iminomethylidene]-6-thionojirimycin skeleton have been synthesized as photoprobes to monitor glycosidase binding. Dansyl, dapoxyl and coumarin fluorophores were appended to the terminal amino group at the N'-substituent by either sulfonamide or amide bridging reaction. All the conjugates behaved as strong (low micromolar to nanomolar) and selective inhibitors of β-glucosidases (almonds and bovine liver) and naringinase, in agreement with the inhibition pattern previously encountered for related iso(thio)urea-type bicyclic sp(2)-iminosugars. The presence of the fluorescent probe allows real-time and continuous monitoring of β-glucosidase inhibition by fluorescence resonance energy transfer (FRET), taking advantage of the intrinsic tryptophan-associated fluorescence of the protein.
新的基于 5N,6S-[N'-(4-氨基丁基)亚氨基甲叉基]-6-硫代乔米林骨架的荧光标记 sp(2)-亚氨基糖已被合成作为光探针来监测糖苷酶结合。丹磺酰基、达泊西汀和香豆素荧光团通过磺酰胺或酰胺桥接反应连接到 N'-取代基的末端氨基上。所有的缀合物都表现出对β-葡萄糖苷酶（杏仁和牛肝）和柚皮苷酶的强（低微摩尔至纳摩尔）和选择性抑制作用，这与先前遇到的相关异（硫）脲型双环 sp(2)-亚氨基糖的抑制模式一致。荧光探针的存在允许通过荧光共振能量转移（FRET）实时和连续监测β-葡萄糖苷酶的抑制，利用蛋白质中固有色氨酸相关的荧光。
