| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2020: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000)
Fiscal Year 2019: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2018: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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| Outline of Final Research Achievements |
In the present study, IDNCL-ER, a detection system for ligand-protein interactions, has been improved to discover the proteins that bind to drugs and drug candidates. To this purpose, we employed β-lactamase, which is known as an enzyme to break β-lactam ring, as a reporter enzyme. A split intein, which can self-catalyze protein trans-splicing (PTS) reaction, was also employed to develop the detection method. In the detection method, a synthetic ligand such as atorvastatin, a known cholesterol-lowering drug, was conjugated with the short peptide tag containing the C-terminal sequences of split intein and β-lactamase. When the synthetic ligand binds to a target protein, PDE6D, bearing N-terminal sequences β-lactamase and split intein, PTS reaction occurs and active β-lactamase is generated. Using this method, we have assessed the interactions between atorvastatin and PDE6D in vitro, and in E. coli.
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