Phase-separated condensate-aided enrichment of biomolecular interactions for high-throughput drug screening in test tubes

J Biol Chem. 2020 Aug 14;295(33):11420-11434. doi: 10.1074/jbc.RA120.012981. Epub 2020 May 27.


Modification-dependent and -independent biomolecular interactions, including protein-protein, protein-DNA/RNA, protein-sugar, and protein-lipid interactions, play crucial roles in all cellular processes. Dysregulation of these biomolecular interactions or malfunction of the associated enzymes results in various diseases; therefore, these interactions and enzymes are attractive targets for therapies. High-throughput screening can greatly facilitate the discovery of drugs for these targets. Here, we describe a biomolecular interaction detection method, called phase-separated condensate-aided enrichment of biomolecular interactions in test tubes (CEBIT). The readout of CEBIT is the selective recruitment of biomolecules into phase-separated condensates harboring their cognate binding partners. We tailored CEBIT to detect various biomolecular interactions and activities of biomolecule-modifying enzymes. Using CEBIT-based high-throughput screening assays, we identified known inhibitors of the p53/MDM2 (MDM2) interaction and of the histone methyltransferase, suppressor of variegation 3-9 homolog 1 (SUV39H1), from a compound library. CEBIT is simple and versatile, and is likely to become a powerful tool for drug discovery and basic biomedical research.

Keywords: CEBIT; SUV39H1; biomolecular interactions; biophysics; high-throughput screening; p53; p53/MDM2 interaction; post-translational modification (PTM); protein complex; protein drug interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Drug Discovery / methods
  • Drug Evaluation, Preclinical / methods*
  • High-Throughput Screening Assays / methods*
  • Humans
  • Methyltransferases / antagonists & inhibitors
  • Methyltransferases / metabolism
  • Phase Transition
  • Protein Interaction Mapping / methods*
  • Protein Interaction Maps / drug effects
  • Proto-Oncogene Proteins c-mdm2 / antagonists & inhibitors
  • Proto-Oncogene Proteins c-mdm2 / metabolism
  • Repressor Proteins / antagonists & inhibitors
  • Repressor Proteins / metabolism
  • Tumor Suppressor Protein p53 / antagonists & inhibitors
  • Tumor Suppressor Protein p53 / metabolism


  • Repressor Proteins
  • Tumor Suppressor Protein p53
  • SUV39H1 protein, human
  • Methyltransferases
  • Proto-Oncogene Proteins c-mdm2