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

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

来源：Journal of the American Chemical Society, 2017, 139(40): 14192-14197.

DOI：10.1021/jacs.7b07352

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 beta-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.

• 出版日期2017-10-11