Identification of a Selective RelA Inhibitor Based on DSE-FRET Screening Methods

Autor: Yoshitomo Shiroma, Go Fujita, Takuya Yamamoto, Ryou-u Takahashi, Ashutosh Kumar, Kam Y. J. Zhang, Akihiro Ito, Hiroyuki Osada, Minoru Yoshida, Hidetoshi Tahara
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Zdroj: International Journal of Molecular Sciences, Vol 21, Iss 23, p 9150 (2020)
Druh dokumentu: article
ISSN: 21239150
1422-0067
1661-6596
DOI: 10.3390/ijms21239150
Popis: Nuclear factor-κB (NF-κB) is an important transcription factor involved in various biological functions, including tumorigenesis. Hence, NF-κB has attracted attention as a target factor for cancer treatment, leading to the development of several inhibitors. However, existing NF-κB 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-κB 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-κB 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–DNA binding. We propose that A55 is a seed compound for RelA-specific inhibition and could be used in clinical applications.
Databáze: Directory of Open Access Journals
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