A Fluorescence Polarization Activity-Based Protein Profiling Assay in the Discovery of Potent, Selective Inhibitors for Human Nonlysosomal Glucosylceramidase

Daniël Lahav; Bing Liu; Richard J. B. H. N. van den Berg; Adrianus M. C. H. van den Nieuwendijk; Tom Wennekes; Amar T. Ghisaidoobe; Imogen Breen; Maria J. Ferraz; Chi-Lin Kuo; Liang Wu; Paul P. Geurink; Huib Ovaa; Gijsbert A. van der Marel; Mario van der Stelt; Rolf G. Boot; Gideon J. Davies; Johannes M. F. G. Aerts; Herman S. Overkleeft

doi:10.1021/jacs.7b07352

A Fluorescence Polarization Activity-Based Protein Profiling Assay in the Discovery of Potent, Selective Inhibitors for Human Nonlysosomal Glucosylceramidase

Daniël Lahav, Bing Liu, Richard J. B. H. N. van den Berg, Adrianus M. C. H. van den Nieuwendijk, Tom Wennekes, Amar T. Ghisaidoobe, Imogen Breen, Maria J. Ferraz, Chi-Lin Kuo, Liang Wu, Paul P. Geurink, Huib Ovaa, Gijsbert A. van der Marel, Mario van der Stelt, Rolf G. Boot, Gideon J. Davies, Johannes M. F. G. Aerts, Herman S. Overkleeft

Chemistry

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

Abstract

Human nonlysosomal glucosylceramidase (GBA2) is one of several enzymes that controls levels of glycolipids and whose activity is linked to several human disease states. There is a major need to design or discover selective GBA2 inhibitors both as chemical tools and as potential therapeutic agents. Here, we describe the development of a fluorescence polarization activity-based protein profiling (FluoPol-ABPP) assay for the rapid identification, from a 350+ library of iminosugars, of GBA2 inhibitors. A focused library is generated based on leads from the FluoPol-ABPP screen and assessed on GBA2 selectivity offset against the other glucosylceramide metabolizing enzymes, glucosylceramide synthase (GCS), lysosomal glucosylceramidase (GBA), and the cytosolic retaining β-glucosidase, GBA3. Our work, yielding potent and selective GBA2 inhibitors, also provides a roadmap for the development of high-throughput assays for identifying retaining glycosidase inhibitors by FluoPol-ABPP on cell extracts containing recombinant, overexpressed glycosidase as the easily accessible enzyme source.

Original language	English
Pages (from-to)	14192-14197
Number of pages	6
Journal	Journal of the American Chemical Society
Volume	139
Issue number	40
Early online date	22 Sept 2017
DOIs	https://doi.org/10.1021/jacs.7b07352
Publication status	Published - 11 Oct 2017

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jacs.7b07352
© 2017 American Chemical Society
Final published version, 1.82 MBLicence: CC BY-NC-ND

Gideon John Davies, FRS, FMedSci
- gideon.daviesyork.acuk
- Chemistry - Professor
Person: Academic

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Lahav, D., Liu, B., van den Berg, R. J. B. H. N., van den Nieuwendijk, A. M. C. H., Wennekes, T., Ghisaidoobe, A. T., Breen, I., Ferraz, M. J., Kuo, C.-L., Wu, L., Geurink, P. P., Ovaa, H., van der Marel, G. A., van der Stelt, M., Boot, R. G., Davies, G. J., Aerts, J. M. F. G., & Overkleeft, H. S. (2017). A Fluorescence Polarization Activity-Based Protein Profiling Assay in the Discovery of Potent, Selective Inhibitors for Human Nonlysosomal Glucosylceramidase. Journal of the American Chemical Society, 139(40), 14192-14197. https://doi.org/10.1021/jacs.7b07352

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title = "A Fluorescence Polarization Activity-Based Protein Profiling Assay in the Discovery of Potent, Selective Inhibitors for Human Nonlysosomal Glucosylceramidase",

abstract = "Human nonlysosomal glucosylceramidase (GBA2) is one of several enzymes that controls levels of glycolipids and whose activity is linked to several human disease states. There is a major need to design or discover selective GBA2 inhibitors both as chemical tools and as potential therapeutic agents. Here, we describe the development of a fluorescence polarization activity-based protein profiling (FluoPol-ABPP) assay for the rapid identification, from a 350+ library of iminosugars, of GBA2 inhibitors. A focused library is generated based on leads from the FluoPol-ABPP screen and assessed on GBA2 selectivity offset against the other glucosylceramide metabolizing enzymes, glucosylceramide synthase (GCS), lysosomal glucosylceramidase (GBA), and the cytosolic retaining β-glucosidase, GBA3. Our work, yielding potent and selective GBA2 inhibitors, also provides a roadmap for the development of high-throughput assays for identifying retaining glycosidase inhibitors by FluoPol-ABPP on cell extracts containing recombinant, overexpressed glycosidase as the easily accessible enzyme source.",

author = "Dani{\"e}l Lahav and Bing Liu and {van den Berg}, {Richard J. B. H. N.} and {van den Nieuwendijk}, {Adrianus M. C. H.} and Tom Wennekes and Ghisaidoobe, {Amar T.} and Imogen Breen and Ferraz, {Maria J.} and Chi-Lin Kuo and Liang Wu and Geurink, {Paul P.} and Huib Ovaa and {van der Marel}, {Gijsbert A.} and {van der Stelt}, Mario and Boot, {Rolf G.} and Davies, {Gideon J.} and Aerts, {Johannes M. F. G.} and Overkleeft, {Herman S.}",

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year = "2017",

month = oct,

day = "11",

doi = "10.1021/jacs.7b07352",

language = "English",

volume = "139",

pages = "14192--14197",

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Lahav, D, Liu, B, van den Berg, RJBHN, van den Nieuwendijk, AMCH, Wennekes, T, Ghisaidoobe, AT, Breen, I, Ferraz, MJ, Kuo, C-L, Wu, L, Geurink, PP, Ovaa, H, van der Marel, GA, van der Stelt, M, Boot, RG, Davies, GJ, Aerts, JMFG & Overkleeft, HS 2017, 'A Fluorescence Polarization Activity-Based Protein Profiling Assay in the Discovery of Potent, Selective Inhibitors for Human Nonlysosomal Glucosylceramidase', Journal of the American Chemical Society, vol. 139, no. 40, pp. 14192-14197. https://doi.org/10.1021/jacs.7b07352

A Fluorescence Polarization Activity-Based Protein Profiling Assay in the Discovery of Potent, Selective Inhibitors for Human Nonlysosomal Glucosylceramidase. / Lahav, Daniël; Liu, Bing; van den Berg, Richard J. B. H. N. et al.
In: Journal of the American Chemical Society, Vol. 139, No. 40, 11.10.2017, p. 14192-14197.

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

TY - JOUR

T1 - A Fluorescence Polarization Activity-Based Protein Profiling Assay in the Discovery of Potent, Selective Inhibitors for Human Nonlysosomal Glucosylceramidase

AU - Lahav, Daniël

AU - Liu, Bing

AU - van den Berg, Richard J. B. H. N.

AU - van den Nieuwendijk, Adrianus M. C. H.

AU - Wennekes, Tom

AU - Ghisaidoobe, Amar T.

AU - Breen, Imogen

AU - Ferraz, Maria J.

AU - Kuo, Chi-Lin

AU - Wu, Liang

AU - Geurink, Paul P.

AU - Ovaa, Huib

AU - van der Marel, Gijsbert A.

AU - van der Stelt, Mario

AU - Boot, Rolf G.

AU - Davies, Gideon J.

AU - Aerts, Johannes M. F. G.

AU - Overkleeft, Herman S.

PY - 2017/10/11

Y1 - 2017/10/11

N2 - Human nonlysosomal glucosylceramidase (GBA2) is one of several enzymes that controls levels of glycolipids and whose activity is linked to several human disease states. There is a major need to design or discover selective GBA2 inhibitors both as chemical tools and as potential therapeutic agents. Here, we describe the development of a fluorescence polarization activity-based protein profiling (FluoPol-ABPP) assay for the rapid identification, from a 350+ library of iminosugars, of GBA2 inhibitors. A focused library is generated based on leads from the FluoPol-ABPP screen and assessed on GBA2 selectivity offset against the other glucosylceramide metabolizing enzymes, glucosylceramide synthase (GCS), lysosomal glucosylceramidase (GBA), and the cytosolic retaining β-glucosidase, GBA3. Our work, yielding potent and selective GBA2 inhibitors, also provides a roadmap for the development of high-throughput assays for identifying retaining glycosidase inhibitors by FluoPol-ABPP on cell extracts containing recombinant, overexpressed glycosidase as the easily accessible enzyme source.

AB - Human nonlysosomal glucosylceramidase (GBA2) is one of several enzymes that controls levels of glycolipids and whose activity is linked to several human disease states. There is a major need to design or discover selective GBA2 inhibitors both as chemical tools and as potential therapeutic agents. Here, we describe the development of a fluorescence polarization activity-based protein profiling (FluoPol-ABPP) assay for the rapid identification, from a 350+ library of iminosugars, of GBA2 inhibitors. A focused library is generated based on leads from the FluoPol-ABPP screen and assessed on GBA2 selectivity offset against the other glucosylceramide metabolizing enzymes, glucosylceramide synthase (GCS), lysosomal glucosylceramidase (GBA), and the cytosolic retaining β-glucosidase, GBA3. Our work, yielding potent and selective GBA2 inhibitors, also provides a roadmap for the development of high-throughput assays for identifying retaining glycosidase inhibitors by FluoPol-ABPP on cell extracts containing recombinant, overexpressed glycosidase as the easily accessible enzyme source.

U2 - 10.1021/jacs.7b07352

DO - 10.1021/jacs.7b07352

M3 - Article

SN - 0002-7863

VL - 139

SP - 14192

EP - 14197

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

IS - 40

ER -

Lahav D, Liu B, van den Berg RJBHN, van den Nieuwendijk AMCH, Wennekes T, Ghisaidoobe AT et al. A Fluorescence Polarization Activity-Based Protein Profiling Assay in the Discovery of Potent, Selective Inhibitors for Human Nonlysosomal Glucosylceramidase. Journal of the American Chemical Society. 2017 Oct 11;139(40):14192-14197. Epub 2017 Sept 22. doi: 10.1021/jacs.7b07352

A Fluorescence Polarization Activity-Based Protein Profiling Assay in the Discovery of Potent, Selective Inhibitors for Human Nonlysosomal Glucosylceramidase

Abstract

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Gideon John Davies, FRS, FMedSci

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