title: A Specific Activity-Based Probe to Monitor Family GH59 Galactosylceramidase, the Enzyme Deficient in Krabbe Disease.
abstract: 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.
authors.affiliation: Department of Biochemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands.|Present address: Institute of Biochemistry, Christian-Albrechts-University of Kiel, Otto-Hahn-Platz 9, 24098, Kiel, Germany.
authors.forename: André R A
authors.initials: AR
authors.lastname: Marques
authors.affiliation: Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, Einsteeinweg 55, 2300 RA, Leiden, The Netherlands.|Present address: Department of Chemistry, Simon Fraser University, 8888 University Drive, Burnaby, V5A 1S6, BC, Canada.
authors.forename: Lianne I
authors.initials: LI
authors.lastname: Willems
authors.affiliation: Department of Biochemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands.
authors.forename: Daniela
authors.initials: D
authors.lastname: Herrera Moro
authors.affiliation: Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, Einsteeinweg 55, 2300 RA, Leiden, The Netherlands.
authors.forename: Bogdan I
authors.initials: BI
authors.lastname: Florea
authors.affiliation: Department of Biochemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands.
authors.forename: Saskia
authors.initials: S
authors.lastname: Scheij
authors.affiliation: Department of Biochemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands.
authors.forename: Roelof
authors.initials: R
authors.lastname: Ottenhoff
authors.affiliation: Department of Biochemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands.
authors.forename: Cindy P A A
authors.initials: CP
authors.lastname: van Roomen
authors.affiliation: Department of Biochemistry, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA, Leiden, The Netherlands.
authors.forename: Marri
authors.initials: M
authors.lastname: Verhoek
authors.affiliation: Department of Biochemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands.
authors.forename: Jessica K
authors.initials: JK
authors.lastname: Nelson
authors.affiliation: Department of Biochemistry, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA, Leiden, The Netherlands.
authors.forename: Wouter W
authors.initials: WW
authors.lastname: Kallemeijn
authors.affiliation: Institute of Organic and Analytical Chemistry, Université D'Orléans, Rue de Chartres, B. P. 6759, 45100, Orléans, France.
authors.forename: Anna
authors.initials: A
authors.lastname: Biela-Banas
authors.affiliation: Institute of Organic and Analytical Chemistry, Université D'Orléans, Rue de Chartres, B. P. 6759, 45100, Orléans, France.
authors.forename: Olivier R
authors.initials: OR
authors.lastname: Martin
authors.affiliation: Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2QQ, UK.
authors.forename: M Begoña
authors.initials: MB
authors.lastname: Cachón-González
authors.affiliation: Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2QQ, UK.
authors.forename: Nee Na
authors.initials: NN
authors.lastname: Kim
authors.affiliation: Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2QQ, UK.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Cox
authors.affiliation: Department of Biochemistry, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA, Leiden, The Netherlands.
authors.forename: Rolf G
authors.initials: RG
authors.lastname: Boot
authors.affiliation: Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, Einsteeinweg 55, 2300 RA, Leiden, The Netherlands.
authors.forename: Herman S
authors.initials: HS
authors.lastname: Overkleeft
authors.affiliation: Department of Biochemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands.|Department of Biochemistry, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA, Leiden, The Netherlands.
authors.forename: Johannes M F G
authors.initials: JM
authors.lastname: Aerts
mesh_terms: D003677:Deficiency Diseases / Q000201:enzymology / Q000235:genetics; D005698:Galactosylceramidase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*; D007965:Leukodystrophy, Globoid Cell / Q000175:diagnosis / Q000201:enzymology* / Q000235:genetics; D016464:Lysosomal Storage Diseases / Q000201:enzymology / Q000235:genetics; D015335:Molecular Probes; D015394:Molecular Structure; D009154:Mutation
chemical_list: D015335:Molecular Probes; D005698:Galactosylceramidase
keywords: Krabbe disease; beta-galactosidase; fluorescent probes; galactosylceramidase; hydrolase