High-throughput strategy to identify inhibitors of histone-binding domains

Methods Enzymol. 2012:512:161-85. doi: 10.1016/B978-0-12-391940-3.00008-1.

Abstract

Many epigenetic proteins recognize the posttranslational modification state of chromatin through their histone-binding domains and thereby recruit nuclear complexes to specific loci within the genome. A number of these domains have been implicated in cancer and other diseases through aberrant binding of chromatin; therefore, identifying small molecules that disrupt histone binding could be a powerful mechanism for disease therapy. We have developed a high-throughput assay for the detection of histone peptide-domain interactions utilizing AlphaScreen technology. Here, we describe how the assay can be first optimized and then performed for high-throughput screening of small molecule-binding inhibitors. We also describe strategies for biochemical validation of small molecules identified.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Binding, Competitive
  • DNA-Binding Proteins / antagonists & inhibitors*
  • DNA-Binding Proteins / chemistry*
  • Data Interpretation, Statistical
  • High-Throughput Screening Assays*
  • Histones / chemical synthesis
  • Histones / chemistry*
  • Humans
  • Molecular Sequence Data
  • Peptide Fragments / chemical synthesis
  • Peptide Fragments / chemistry*
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Signal-To-Noise Ratio
  • Small Molecule Libraries
  • User-Computer Interface

Substances

  • DNA-Binding Proteins
  • Histones
  • Peptide Fragments
  • Small Molecule Libraries