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A Fluorescence Polarization Activity-Based Protein Profiling Assay in the Discovery of Potent, Selective Inhibitors for Human Nonlysosomal Glucosylceramidase.
- Source :
-
Journal of the American Chemical Society . 10/11/2017, Vol. 139 Issue 40, p14192-14197. 6p. - Publication Year :
- 2017
-
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. [ABSTRACT FROM AUTHOR]
- Subjects :
- *FLUORESCENCE polarization immunoassay
*PROTEINS
*GLYCOLIPIDS
*GLYCOSIDASES
*ENZYMES
Subjects
Details
- Language :
- English
- ISSN :
- 00027863
- Volume :
- 139
- Issue :
- 40
- Database :
- Academic Search Index
- Journal :
- Journal of the American Chemical Society
- Publication Type :
- Academic Journal
- Accession number :
- 125688910
- Full Text :
- https://doi.org/10.1021/jacs.7b07352