| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Outline of Final Research Achievements |
In this study, we used a site-specific spin-labeled electron spin resonance (SDSL-ESR) method. An arbitrary position on KaiB was substituted by Cys residue and a spin label (MTSSL) was introduced into Cys residue. The KaiA interaction site on KaiB was determined by detecting released MTSSL from KaiB when the labeled KaiB and KaiA C-terminal domain proteins were mixed and reacted at physiological temperature. Next, ESR measurements were performed in the presence of the KaiABC mixture to verify when the spin label was released. As a result, the spin label was released 12 hours after the start of the reaction. This result indicates that the KaiA-KaiB interaction starts around 12 hours.
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