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 B T Ghisaidoobe; Imogen Breen; Maria Joao 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 B T Ghisaidoobe, Imogen Breen, Maria Joao 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

Structural Biology Laboratory, Department of Chemistry, The University of York , York YO10 5DD, United Kingdom.
Department of Chemical Immunology, Leiden University Medical Center , Einthovenweg 20, 2333 ZC Leiden, The Netherlands.

Research output: Contribution to journal › Article › Academic › 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
DOIs	https://doi.org/10.1021/jacs.7b07352
Publication status	Published - 2017

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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. B. 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

@article{66e9e14ef4b84bc59e43000c84105e46,

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 B T} and Imogen Breen and Ferraz, {Maria Joao} 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.}",

year = "2017",

doi = "10.1021/jacs.7b07352",

language = "English",

volume = "139",

pages = "14192--14197",

journal = "Journal of the American Chemical Society",

issn = "0002-7863",

publisher = "American Chemical Society",

number = "40",

}

Lahav, D, Liu, B, van den Berg, RJBHN, van den Nieuwendijk, AMCH, Wennekes, T, Ghisaidoobe, ABT, 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, 2017, p. 14192-14197.

Research output: Contribution to journal › Article › Academic › 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 B T

AU - Breen, Imogen

AU - Ferraz, Maria Joao

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

Y1 - 2017

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

C2 - 28937220

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 -

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

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