A new set of assays for the discovery of aminoacyl-tRNA synthetase inhibitors

Methods. 2017 Jan 15:113:34-45. doi: 10.1016/j.ymeth.2016.10.011. Epub 2016 Oct 29.

Abstract

Current biochemical methods available to monitor the activity of aminoacyl-tRNA synthetases (ARS) are ill-suited to high-throughput screening approaches for the identification of small-molecule inhibitors of these enzymes. In an attempt to improve the limitations of current assays we have developed a suite of new methods designed to streamline the discovery of new ARS antagonists. This set of assays includes approaches to monitor ARS activity in vitro, in human cells, and in bacteria. They are applicable to several ARSs from any given organism, can be easily adapted to very high-throughput set-ups, and allow for a multi-factorial selection of drug candidates.

Keywords: Aminoacyl-tRNA synthetase; Bioluminescence; Enzymatic assay; High-throughput screening; Luciferase; tRNA.

MeSH terms

  • Amino Acids / metabolism
  • Amino Acyl-tRNA Synthetases / antagonists & inhibitors*
  • Amino Acyl-tRNA Synthetases / genetics
  • Amino Acyl-tRNA Synthetases / metabolism
  • Arabidopsis / enzymology
  • Arabidopsis / genetics
  • Drug Discovery
  • Enzyme Assays
  • Enzyme Inhibitors / pharmacology*
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Genes, Reporter
  • High-Throughput Screening Assays*
  • Humans
  • Luciferases / genetics
  • Luciferases / metabolism
  • Luminescent Measurements / methods
  • Methicillin-Resistant Staphylococcus aureus / enzymology
  • Methicillin-Resistant Staphylococcus aureus / genetics
  • RNA, Transfer, Amino Acid-Specific / genetics*
  • RNA, Transfer, Amino Acid-Specific / metabolism
  • Small Molecule Libraries / pharmacology*
  • Transfer RNA Aminoacylation*

Substances

  • Amino Acids
  • Enzyme Inhibitors
  • RNA, Transfer, Amino Acid-Specific
  • Small Molecule Libraries
  • Luciferases
  • Amino Acyl-tRNA Synthetases