biostudies-literatureJiang JEuropean Research CouncilBiotechnology and Biological Sciences Research Council2782-falsehttps://www.ebi.ac.uk/biostudies/studies/S-EPMC5654414biostudies-literatureUnknown6(5)GH29 α-l-fucosidases catalyze the hydrolysis of α-l-fucosidic linkages. Deficiency in human lysosomal α-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 <i>in vitro</i> and <i>in vivo</i> labeling of GH29 α-l-fucosidases from bacteria, mice and man. Crystallographic analysis on bacterial α-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 α-l-fucosidase inhibitor from eight configurational isomers.Chemical science&lt;i&gt;In vitro&lt;/i&gt; and &lt;i&gt;in vivo&lt;/i&gt; comparative and competitive activity-based protein profiling of GH29 α-l-fucosidases.PMC56544143229421094267van den Nieuwendijk AMCHJiang JDonker-Koopman WEFolch ECScheij SSchurmann MVerhoek MWright DWvan den Elst HDavies GJKallemeijn WWFlorea BILi Nvan der Marel GACodee JDCOverkleeft HSBoot RGAerts JMFGRohde VCMink Dfalse&lt;i&gt;In vitro&lt;/i&gt; and &lt;i&gt;in vivo&lt;/i&gt; comparative and competitive activity-based protein profiling of GH29 α-l-fucosidases.GH29 α-l-fucosidases catalyze the hydrolysis of α-l-fucosidic linkages. Deficiency in human lysosomal α-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 <i>in vitro</i> and <i>in vivo</i> labeling of GH29 α-l-fucosidases from bacteria, mice and man. Crystallographic analysis on bacterial α-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 α-l-fucosidase inhibitor from eight configurational isomers.2015-01-01T00:00:00Z2015 May2026-04-30T09:19:19.083Z2019-03-26T23:02:45ZS-EPMC56544142914268110.1039/c4sc03739a