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In vivo inactivation of glycosidases by conduritol B epoxide and cyclophellitol as revealed by activity‐based protein profiling
Author(s) -
Kuo ChiLin,
Kallemeijn Wouter W.,
Lelieveld Lindsey T.,
Mirzaian Mina,
Zoutendijk Iris,
Vardi Ayelet,
Futerman Anthony H.,
Meijer Annemarie H.,
Spaink Herman P.,
Overkleeft Herman S.,
Aerts Johannes M.F.G.,
Artola Marta
Publication year - 2019
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.14744
Subject(s) - glucocerebrosidase , in vivo , enzyme , biochemistry , glycoside hydrolase , chemistry , hydrolase , biology , genetics
Glucocerebrosidase ( GBA ) is a lysosomal β‐glucosidase‐degrading glucosylceramide. Its deficiency causes Gaucher disease ( GD ), a common lysosomal storage disorder. Carrying a genetic abnormality in GBA constitutes at present the largest genetic risk factor for Parkinson's disease ( PD ). Conduritol B epoxide ( CBE ), a mechanism‐based irreversible inhibitor of GBA , is used to generate cell and animal models for investigations on GD and PD . However, CBE may have additional glycosidase targets besides GBA . Here, we present the first in vivo target engagement study for CBE , employing a suite of activity‐based probes to visualize catalytic pocket occupancy of candidate off‐target glycosidases. Only at significantly higher CBE concentrations, nonlysosomal glucosylceramidase ( GBA 2) and lysosomal α‐glucosidase were identified as major off‐targets in cells and zebrafish larvae. A tight, but acceptable window for selective inhibition of GBA in the brain of mice was observed. On the other hand, cyclophellitol, a closer glucose mimic, was found to inactivate with equal affinity GBA and GBA 2 and therefore is not suitable to generate genuine GD ‐like models. Enzymes Glucocerebrosidase ( EC 3.2.1.45 ), nonlysosomal β‐glucocerebrosidase ( EC 3.2.1.45 ); cytosolic β‐glucosidase ( EC 3.2.1.21 ); α‐glucosidases ( EC 3.2.1.20 ); β‐glucuronidase ( EC 3.2.1.31 ).