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

doi:10.1002/cbic.201600561

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

Chemistry

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Pages (from-to)	402-412
Number of pages	11
Journal	Chembiochem
Volume	18
Issue number	4
Early online date	20 Dec 2016
DOIs	https://doi.org/10.1002/cbic.201600561
Publication status	Published - 31 Jan 2017

Bibliographical note

This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details

Keywords

beta-galactosidase
fluorescent probes
galactosylceramidase
hydrolase
Krabbe disease

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10.1002/cbic.201600561

Marques, Willems, et al. Chembiochem 2017, Accepted manuscript
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Marques, A. R. A., Willems, L. I., Herrera Moro, D., Florea, B. I., Scheij, S., Ottenhoff, R., van Roomen, C. P. A. A., Verhoek, M., Nelson, J. K., Kallemeijn, W. W., Biela-Banas, A., Martin, O. R., Cachón‐González , M. B., Kim, N. N., Cox, T. M., Boot, R. G., Overkleeft, H. S., & Aerts, J. M. F. G. (2017). A Specific Activity-Based Probe to Monitor Family GH59 Galactosylceramidase, the Enzyme Deficient in Krabbe Disease. Chembiochem, 18(4), 402-412. https://doi.org/10.1002/cbic.201600561

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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.",

keywords = "beta-galactosidase, fluorescent probes, galactosylceramidase, hydrolase, Krabbe disease",

author = "Marques, {Andr{\'e} R. A.} and Willems, {Lianne I.} and {Herrera Moro}, Daniela and Florea, {Bogdan I.} and Saskia Scheij and Roelof Ottenhoff and {van Roomen}, {Cindy P.A.A.} and Marri Verhoek and Nelson, {Jessica K.} and Kallemeijn, {Wouter W.} and Anna Biela-Banas and Martin, {Olivier R.} and Cach{\'o}n‐Gonz{\'a}lez, {M. Bego{\~n}a} and Kim, {Nee Na} and Cox, {Timothy M.} and Boot, {Rolf G.} and Overkleeft, {Herman S.} and Aerts, {Johannes M F G}",

note = "This is an author-produced version of the published paper. Uploaded in accordance with the publisher{\textquoteright}s self-archiving policy. Further copying may not be permitted; contact the publisher for details",

year = "2017",

month = jan,

day = "31",

doi = "10.1002/cbic.201600561",

language = "English",

volume = "18",

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Marques, ARA, Willems, LI, Herrera Moro, D, Florea, BI, Scheij, S, Ottenhoff, R, van Roomen, CPAA, Verhoek, M, Nelson, JK, Kallemeijn, WW, Biela-Banas, A, Martin, OR, Cachón‐González , MB, Kim, NN, Cox, TM, Boot, RG, Overkleeft, HS & Aerts, JMFG 2017, 'A Specific Activity-Based Probe to Monitor Family GH59 Galactosylceramidase, the Enzyme Deficient in Krabbe Disease', Chembiochem, vol. 18, no. 4, pp. 402-412. https://doi.org/10.1002/cbic.201600561

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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 P.A.A.

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

N1 - This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details

PY - 2017/1/31

Y1 - 2017/1/31

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.

KW - beta-galactosidase

KW - fluorescent probes

KW - galactosylceramidase

KW - hydrolase

KW - Krabbe disease

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DO - 10.1002/cbic.201600561

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SN - 1439-4227

VL - 18

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JO - Chembiochem

JF - Chembiochem

IS - 4

ER -

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

Abstract

Bibliographical note

Keywords

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