In vitro and in vivo comparative and competitive activity-based protein profiling of GH29 alpha-L-fucosidases

摘要

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