Applications for activity-based probes in biomedical research on glycosidases

Kuo, C.L. (2019)

Doctoral Thesis

Lysosomal glycosidases are acid hydrolases that fragment glycoconjugates in lysosomes. Their inherited deficiency in human is the cause of a number of lysosomal storage disorders (LSDs), showing characteristic lysosomal accumulation of undegraded glycoconjugates. In the past, activity-based probes (ABPs) based on cyclophellitol or cyclophellitol aziridine scaffold have emerged as powerful tools enabling sensitive quantification of a number of lysosomal glycosidases in extracts of cells and tissue, as well as in intact cells. This thesis describes the characterization of several novel ABP classes targeting α-glucosidase, β-glucuronidase, α-L-iduronidase, α-mannosidase, β-mannosidase, and β-galactosidase, as well as a broad scale of applications for ABPs in LSD research. Novel glucocerebrosidase inhibitors based on the cyclophellitol scaffold are also described, which are brain-permeable, selective, and potently inactivate the enzyme in adult zebrafish. Additionally, a protocol for gel-based and microscopy-based detection of glucocerebrosidase is described.

• Activity-based probe
• Activity-based protein profiling
• Lysosomal storage disorders
• Cyclophellitol
• Glucocerebrosidase
• Alpha-glucosidase
• Mannosidase
• Beta-galactosidase
• Iduronidase
• Beta-glucuronidase

• All authors: Kuo, C.L.
• Supervisor: Aerts, J.M.F.G.; Overkleeft, H.S.
• Committee: Brouwer, J.; Stelt, M. van der; Meijer, A.H.; Saftig, P.; Wachten, D.
• Qualification: Doctor (dr.)
• Awarding Institution: Institute of Chemistry (LIC), Faculty of Science, Leiden University
• Date: 2019-09-10
• ISBN (print): 9789463754453