Activity-Based Probes for Glycosidases: Profiling and Other Applications.

Autor:	Kuo CL; Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands., van Meel E; Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands., Kytidou K; Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands., Kallemeijn WW; Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands., Witte M; Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands., Overkleeft HS; Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands., Artola ME; Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands., Aerts JM; Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands. Electronic address: j.m.f.g.aerts@lic.leidenuniv.nl.
Jazyk:	angličtina
Zdroj:	Methods in enzymology [Methods Enzymol] 2018; Vol. 598, pp. 217-235. Date of Electronic Publication: 2017 Dec 08.
DOI:	10.1016/bs.mie.2017.06.039
Abstrakt:	Glycosidases mediate the fragmentation of glycoconjugates in the body, including the vital recycling of endogenous molecules. Several inherited diseases in man concern deficiencies in lysosomal glycosidases degrading glycosphingolipids. Prominent is Gaucher disease caused by an impaired lysosomal β-glucosidase (glucocerebrosidase, GBA) and resulting in pathological lysosomal storage of glucosylceramide (glucocerebroside) in tissue macrophages. GBA is a retaining glucosidase with a characteristic glycosyl-enzyme intermediate formed during catalysis. Using the natural suicide inhibitor cyclophellitol as a lead, we developed mechanism-based irreversible inhibitors of GBA equipped with a fluorescent reporter. These reagents covalently link to the catalytic nucleophile residue of GBA and permit specific and sensitive visualization of active enzyme molecules. The amphiphilic activity-based probes (ABPs) allow in situ detection of active GBA in cells and organisms. Furthermore, they may be used to biochemically confirm the diagnosis of Gaucher disease and they might assist in screening for small compounds interacting with the catalytic pocket. While the focus of this chapter is ABPs for β-glucosidases and Gaucher disease, the described concept has meanwhile been extended to other retaining glycosidases and related disease conditions as well. (© 2018 Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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