In vitro and in vivo comparative and competitive activity-based protein profiling of GH29 α-L-fucosidases
In vitro and in vivo comparative and competitive activity-based protein profiling of GH29 α-L-fucosidases
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DOI：
10.1039/c4sc03739a
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发表时间：
2015-01-01
期刊：
影响因子：
8.4
通讯作者：
Overkleeft, Herman S.
中科院分区：
文献类型：
--
作者：
Jiang, Jianbing;Kallemeijn, Wouter W.;Overkleeft, Herman S.
GH29 α-L-岩藻糖苷酶催化 α-L-岩藻糖苷键的水解。人类溶酶体 α-L-岩藻糖苷酶 (FUCA1) 缺乏会导致隐性遗传性疾病——岩藻糖苷中毒。在此，我们描述了吡喃岩藻糖配置的环己醇氮丙啶作为基于活性的探针（ABP）的开发，用于选择性体外和体内标记来自细菌、小鼠和人类的GH29 α-L-岩藻糖苷酶。对细菌 α-L-岩藻糖苷酶的晶体分析证实，ABP 通过活性位点亲核试剂的共价修饰发挥作用。基于竞争活性的蛋白质分析将 L-岩藻糖野尻霉素确定为八种构型异构体中的单一 GH29 α-L-岩藻糖苷酶抑制剂。
GH29 alpha-L-fucosidases catalyze the hydrolysis of alpha-L-fucosidic linkages. Deficiency in human lysosomal alpha-L-fucosidase (FUCA1) leads to the recessively inherited disorder, fucosidosis. Herein we describe the development of fucopyranose-configured cyclophellitol aziridines as activity-based probes (ABPs) for selective in vitro and in vivo labeling of GH29 alpha-L-fucosidases from bacteria, mice and man. Crystallographic analysis on bacterial alpha-L-fucosidase confirms that the ABPs act by covalent modification of the active site nucleophile. Competitive activity-based protein profiling identified L-fuconojirimycin as the single GH29 alpha-L-fucosidase inhibitor from eight configurational isomers.