An Economical High-Throughput "FP-Tag" Assay for Screening Glycosyltransferase Inhibitors*

Chembiochem. 2021 Apr 16;22(8):1391-1395. doi: 10.1002/cbic.202000746. Epub 2021 Jan 21.

Abstract

O-GlcNAc transferase (OGT) is involved in many cellular processes, and selective OGT inhibitors are valuable tools to investigate O-GlcNAcylation functions, and could potentially lead to therapeutics. However, high-throughput OGT assays that are suitable for large-scale HTS and can identify inhibitors targeting both acceptor, donor sites, and allosteric binding-sites are still lacking. Here, we report the development of a high-throughput "FP-Tag" OGT assay with bovine serum albumin (BSA) as a low-cost and superior "FP-Tag". With this assay, 2-methyleurotinone was identified as a low-micromolar OGT inhibitor. This type of assay with BSA as "FP-Tag" would find more applications with other glycosyltransferases.

Keywords: bovine serum albumin; fluorescence polarization assay; glycosyltransferases; high-throughput screening; inhibitors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkynes / chemistry
  • Animals
  • Biotin / chemistry
  • Cattle
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / pharmacology*
  • Fluorescent Dyes / chemistry
  • High-Throughput Screening Assays*
  • Humans
  • Molecular Structure
  • N-Acetylglucosaminyltransferases / antagonists & inhibitors*
  • N-Acetylglucosaminyltransferases / metabolism
  • Serum Albumin, Bovine / chemistry
  • Streptavidin / chemistry

Substances

  • Alkynes
  • Enzyme Inhibitors
  • Fluorescent Dyes
  • Serum Albumin, Bovine
  • Biotin
  • Streptavidin
  • N-Acetylglucosaminyltransferases
  • O-GlcNAc transferase