A specific activity-based probe to monitor family GH59 galactosylceramidase - the enzyme deficient in Krabbe disease
| Autor: | Daniela Herrera Moro, Bogdan I. Florea, Timothy M. Cox, Olivier R. Martin, Marri Verhoek, Saskia Scheij, André R. A. Marques, Nee Na Kim, Wouter W. Kallemeijn, Rolf G. Boot, Jessica K. Nelson, Herman S. Overkleeft, Lianne I. Willems, Roelof Ottenhoff, Johannes M. F. G. Aerts, Cindy P. A. A. van Roomen, M. Begoña Cachón-González, Anna Biela-Banas |
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| Přispěvatelé: | Medical Biochemistry, Graduate School, Amsterdam Gastroenterology Endocrinology Metabolism, Amsterdam Cardiovascular Sciences, ACS - Atherosclerosis & ischemic syndromes, ACS - Diabetes & metabolism |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
010402 general chemistry medicine.disease_cause 01 natural sciences Biochemistry 03 medical and health sciences Galactosylceramidase Hydrolase medicine Beta-galactosidase Molecular Biology chemistry.chemical_classification Mutation biology Molecular Structure Organic Chemistry medicine.disease 0104 chemical sciences Leukodystrophy Globoid Cell Lysosomal Storage Diseases 030104 developmental biology Enzyme chemistry Molecular Probes biology.protein Krabbe disease Molecular Medicine Specific activity Molecular probe Deficiency Diseases |
| Zdroj: | Chembiochem, 18(4), 402-412. Wiley-VCH Verlag |
| ISSN: | 1439-7633 1439-4227 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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