Publication

Activity-Based Probes for Glycosidases: Profiling and Other Applications

Kuo, C-L., van Meel, E., Kytidou, K., Kallemeijn, W. W., Witte, M., Overkleeft, H. S., Artola, M. E. & Aerts, J. M., 2018, CHEMICAL GLYCOBIOLOGY, PT B: MONITORING GLYCANS AND THEIR INTERACTIONS. Imperiali, B. (ed.). Academic Press, p. 217-235 19 p. (Methods in Enzymology; vol. 598).

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DOI

  • Chi-Lin Kuo
  • Eline van Meel
  • Kassiani Kytidou
  • Wouter Willem Kallemeijn
  • Martin Witte
  • Herman Stephen Overkleeft
  • Marta Elena Artola
  • Johannes Maria Aerts

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 beta-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 beta-glucosidases and Gaucher disease, the described concept has meanwhile been extended to other retaining glycosidases and related disease conditions as well.

Original languageEnglish
Title of host publicationCHEMICAL GLYCOBIOLOGY, PT B: MONITORING GLYCANS AND THEIR INTERACTIONS
EditorsB Imperiali
PublisherAcademic Press
Pages217-235
Number of pages19
Publication statusPublished - 2018
Externally publishedYes

Publication series

NameMethods in Enzymology
PublisherELSEVIER ACADEMIC PRESS INC
Volume598
ISSN (Print)0076-6879

    Keywords

  • GAUCHER-DISEASE, IN-SITU, GLUCOCEREBROSIDASE, ENZYME, CYCLOPHELLITOL, INHIBITORS, SPECTRUM, GLUCOSIDASES, MECHANISMS, PHENOTYPE

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