A Specific Activity-Based Probe to Monitor Family GH59 Galactosylceramidase, the Enzyme Deficient in Krabbe Disease

André R. A. Marques, Lianne I. Willems, Daniela Herrera Moro, Bogdan I. Florea, Saskia Scheij, Roelof Ottenhoff, Cindy P.A.A. van Roomen, Marri Verhoek, Jessica K. Nelson, Wouter W. Kallemeijn, Anna Biela-Banas, Olivier R. Martin, M. Begoña Cachón‐González , Nee Na Kim, Timothy M. Cox, Rolf G. Boot, Herman S. Overkleeft*, Johannes M F G Aerts

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish
Pages (from-to)402-412
Number of pages11
Issue number4
Early online date20 Dec 2016
Publication statusPublished - 31 Jan 2017

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  • beta-galactosidase
  • fluorescent probes
  • galactosylceramidase
  • hydrolase
  • Krabbe disease

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