Activity-Based Probes for Glycosidases: Profiling and Other ApplicationsKuo, 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).
Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review
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.
|Title of host publication||CHEMICAL GLYCOBIOLOGY, PT B: MONITORING GLYCANS AND THEIR INTERACTIONS|
|Number of pages||19|
|Publication status||Published - 2018|
|Name||Methods in Enzymology|
|Publisher||ELSEVIER ACADEMIC PRESS INC|
- GAUCHER-DISEASE, IN-SITU, GLUCOCEREBROSIDASE, ENZYME, CYCLOPHELLITOL, INHIBITORS, SPECTRUM, GLUCOSIDASES, MECHANISMS, PHENOTYPE