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.
Medical Biochemistr