Abstract
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.
© 2020 Zhou et al.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Drug Discovery / methods
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Drug Evaluation, Preclinical / methods*
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High-Throughput Screening Assays / methods*
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Humans
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Methyltransferases / antagonists & inhibitors
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Methyltransferases / metabolism
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Phase Transition
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Protein Interaction Mapping / methods*
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Protein Interaction Maps / drug effects
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Proto-Oncogene Proteins c-mdm2 / antagonists & inhibitors
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Proto-Oncogene Proteins c-mdm2 / metabolism
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Repressor Proteins / antagonists & inhibitors
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Repressor Proteins / metabolism
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Tumor Suppressor Protein p53 / antagonists & inhibitors
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Tumor Suppressor Protein p53 / metabolism
Substances
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Repressor Proteins
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Tumor Suppressor Protein p53
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SUV39H1 protein, human
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Methyltransferases
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Proto-Oncogene Proteins c-mdm2