TY - JOUR
T1 - A Specific Activity-Based Probe to Monitor Family GH59 Galactosylceramidase, the Enzyme Deficient in Krabbe Disease
AU - Marques, André R. A.
AU - Willems, Lianne I.
AU - Herrera Moro, Daniela
AU - Florea, Bogdan I.
AU - Scheij, Saskia
AU - Ottenhoff, Roelof
AU - van Roomen, Cindy
AU - Verhoek, Marri
AU - Nelson, Jessica K.
AU - Kallemeijn, Wouter W.
AU - Biela-Banas, Anna
AU - Martin, Olivier R.
AU - Cachón-González, M. Begoña
AU - Kim, Nee Na
AU - Cox, Timothy M.
AU - Boot, Rolf G.
AU - Overkleeft, Herman S.
AU - Aerts, Johannes M. F. G.
PY - 2017
Y1 - 2017
N2 - Galactosylceramidase (GALC) is the lysosomal β-galactosidase responsible for the hydrolysis of galactosylceramide. Inherited deficiency in GALC causes Krabbe disease, a devastating neurological disorder characterized by accumulation of galactosylceramide and its deacylated counterpart, the toxic sphingoid base galactosylsphingosine (psychosine). We report the design and application of a fluorescently tagged activity-based probe (ABP) for the sensitive and specific labeling of active GALC molecules from various species. The probe consists of a β-galactopyranose-configured cyclophellitol-epoxide core, conferring specificity for GALC, equipped with a BODIPY fluorophore at C6 that allows visualization of active enzyme in cells and tissues. Detection of residual GALC in patient fibroblasts holds great promise for laboratory diagnosis of Krabbe disease. We further describe a procedure for in situ imaging of active GALC in murine brain by intra-cerebroventricular infusion of the ABP. In conclusion, this GALC-specific ABP should find broad applications in diagnosis, drug development, and evaluation of therapy for Krabbe disease
AB - Galactosylceramidase (GALC) is the lysosomal β-galactosidase responsible for the hydrolysis of galactosylceramide. Inherited deficiency in GALC causes Krabbe disease, a devastating neurological disorder characterized by accumulation of galactosylceramide and its deacylated counterpart, the toxic sphingoid base galactosylsphingosine (psychosine). We report the design and application of a fluorescently tagged activity-based probe (ABP) for the sensitive and specific labeling of active GALC molecules from various species. The probe consists of a β-galactopyranose-configured cyclophellitol-epoxide core, conferring specificity for GALC, equipped with a BODIPY fluorophore at C6 that allows visualization of active enzyme in cells and tissues. Detection of residual GALC in patient fibroblasts holds great promise for laboratory diagnosis of Krabbe disease. We further describe a procedure for in situ imaging of active GALC in murine brain by intra-cerebroventricular infusion of the ABP. In conclusion, this GALC-specific ABP should find broad applications in diagnosis, drug development, and evaluation of therapy for Krabbe disease
U2 - https://doi.org/10.1002/cbic.201600561
DO - https://doi.org/10.1002/cbic.201600561
M3 - Article
C2 - 28000364
SN - 1439-4227
VL - 18
SP - 402
EP - 412
JO - Chembiochem
JF - Chembiochem
IS - 4
ER -