Related Experiment Video
Updated: Apr 17, 2026
08:34
A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening
Published on: October 16, 2015
10.7K
A rapid and sensitive high-throughput screening method to identify compounds targeting protein-nucleic acids
Nicole Alonso1, Roboan Guillen1, Jeremy W Chambers2
1Biomolecular Sciences Institute, Florida International University, 11200 SW 8th Street, Miami, FL 33199, USA Department of Chemistry & Biochemistry, Florida International University, 11200 SW 8th Street, FL 33199, USA.
Nucleic Acids Research
|February 6, 2015
Summary
Researchers developed a high-throughput screening method to find drugs targeting DNA-binding and RNA-binding proteins. This assay successfully identified inhibitors of high-mobility-group protein AT-hook 2 (HMGA2), impacting cell differentiation.
Area of Science:
- Biochemistry
- Molecular Biology
- Drug Discovery
Background:
- DNA-binding and RNA-binding proteins are challenging therapeutic targets due to difficulties in identifying small molecule inhibitors.
- Existing methods for screening inhibitors of protein-nucleic acid interactions are often inefficient.
Purpose of the Study:
- To develop a rapid, sensitive, and broadly applicable high-throughput screening (HTS) assay for identifying modulators of DNA-binding and RNA-binding proteins.
- To validate the HTS assay using a known protein-DNA interaction and subsequently identify novel modulators.
Main Methods:
- Development and validation of protein-DNA interaction enzyme-linked immunosorbent assay (PDI-ELISA) and protein-RNA interaction enzyme-linked immunosorbent assay (PRI-ELISA).
- Utilized PDI-ELISA to screen a library of 29 DNA-binding compounds against mammalian high-mobility-group protein AT-hook 2 (HMGA2).
- Assessed the biological effect of identified inhibitors on 3T3-L1 pre-adipocyte differentiation.
Main Results:
- The PDI-ELISA method was successfully validated using HMGA2 and its known inhibitor, netropsin.
- The HTS screen identified several novel inhibitors and one activator of HMGA2-DNA interactions.
- Netropsin, an HMGA2-DNA interaction inhibitor, significantly inhibited 3T3-L1 cell differentiation into adipocytes.
Conclusions:
- The developed PDI-ELISA and PRI-ELISA methods provide a robust platform for HTS of compounds targeting protein-nucleic acid interactions.
- This approach can accelerate the discovery of novel therapeutic agents for targets previously considered 'undruggable'.
- Inhibition of HMGA2-DNA binding by netropsin demonstrates a potential therapeutic strategy for modulating cell differentiation.
