Identification of a Selective RelA Inhibitor Based on DSE-FRET Screening Methods
| Autor: | Ashutosh Kumar, Kam Y. J. Zhang, Hidetoshi Tahara, Ryou‐u Takahashi, Akihiro Ito, Yoshitomo Shiroma, Takuya Yamamoto, Go Fujita, Hiroyuki Osada, Minoru Yoshida |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Models
Molecular 0301 basic medicine Nuclear Theory Drug Evaluation Preclinical Molecular Conformation NF-κB lcsh:Chemistry chemistry.chemical_compound 0302 clinical medicine Fluorescence Resonance Energy Transfer Nuclear Experiment lcsh:QH301-705.5 Spectroscopy Molecular Structure Chemistry RELB General Medicine Computer Science Applications Molecular Docking Simulation Mathematics::Logic Biochemistry 030220 oncology & carcinogenesis Protein Binding High-throughput screening Protein subunit DNA-binding protein Molecular Dynamics Simulation high-throughput screening Article Catalysis drug discovery Inorganic Chemistry Structure-Activity Relationship 03 medical and health sciences Cell Line Tumor Humans Electrophoretic mobility shift assay Luciferase Physical and Theoretical Chemistry Molecular Biology Transcription factor Binding Sites Organic Chemistry Transcription Factor RelA DNA High-Throughput Screening Assays 030104 developmental biology Förster resonance energy transfer lcsh:Biology (General) lcsh:QD1-999 |
| Zdroj: | International Journal of Molecular Sciences Volume 21 Issue 23 International Journal of Molecular Sciences, Vol 21, Iss 9150, p 9150 (2020) |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms21239150 |
| Popis: | Nuclear factor-&kappa B (NF-&kappa B) is an important transcription factor involved in various biological functions, including tumorigenesis. Hence, NF-&kappa B has attracted attention as a target factor for cancer treatment, leading to the development of several inhibitors. However, existing NF-&kappa B inhibitors do not discriminate between its subunits, namely, RelA, RelB, cRel, p50, and p52. Conventional methods used to evaluate interactions between transcription factors and DNA, such as electrophoretic mobility shift assay and luciferase assays, are unsuitable for high-throughput screening (HTS) and cannot distinguish NF-&kappa B subunits. We developed a HTS method named DNA strand exchange fluorescence resonance energy transfer (DSE-FRET). This assay is suitable for HTS and can discriminate a NF-&kappa B subunit. Using DSE-FRET, we searched for RelA-specific inhibitors and verified RelA inhibition for 32,955 compounds. The compound A55 (2-(3-carbamoyl-6-hydroxy-4-methyl-2-oxopyridin-1(2H)-yl) acetic acid) selectively inhibited RelA&ndash DNA binding. We propose that A55 is a seed compound for RelA-specific inhibition and could be used in clinical applications. |
| Databáze: | OpenAIRE |
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