1. Identification of novel compounds that inhibit SnRK2 kinase activity by high-throughput screening
- Author
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Taishi Umezawa, Shoko Matsuoka, Hirotatsu Kojima, Karin Sato, Riyo Maruki-Imamura, Takayoshi Okabe, and Yoshiteru Noutoshi
- Subjects
0301 basic medicine ,High-throughput screening ,Biophysics ,Arabidopsis ,Protein Serine-Threonine Kinases ,Genes, Plant ,Biochemistry ,Chemical library ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Gene Expression Regulation, Plant ,Kinase activity ,Phosphorylation ,Protein kinase A ,Molecular Biology ,Abscisic acid ,Protein Kinase Inhibitors ,biology ,Kinase ,Arabidopsis Proteins ,fungi ,food and beverages ,Cell Biology ,biology.organism_classification ,Recombinant Proteins ,High-Throughput Screening Assays ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,Abscisic Acid - Abstract
Abscisic acid (ABA) is a major phytohormone that regulates abiotic stress responses and development. SNF1-rerated protein kinase 2 (SnRK2) is a key regulator of ABA signaling. To isolate compounds which directly affect SnRK2 activity, we optimized a fluorescence-based system for high-throughput screening (HTS) of SnRK2 kinase regulators. Using this system, we screened a chemical library consisting of 16,000 compounds and identified ten compounds (INH1-10) as potential SnRK2 inhibitors. Further characterization of these compounds by in vitro phosphorylation assays confirmed that three of the ten compounds were SnRK2-specific kinase inhibitors. In contrast, seven of ten compounds inhibited ABA-responsive gene expression in Arabidopsis cells. From these results, INH1 was identified as a SnRK2-specific inhibitor in vitro and in vivo. We propose that INH1 could be a lead compound of chemical tools for studying ABA responses in various plant species.
- Published
- 2020