| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
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| Outline of Final Research Achievements |
Voltage-gated ion channels have multi subunits and cooperative gating. However, the mechanism of the cooperativity is not well understood. Voltage-gated proton channel (Hv1/VSOP) assembles as a dimeric stoichiometry to regulate the cooperative gating. I investigated real-time monitoring of structural changes of Hv1/VSOP to reveal the mechanism of subunit cooperativity. Cy3- or Cy5-labeled Hv1/VSOP induced fluorescent change under depolarizing pulse (Cy3, Cy5 : classical environmental-sensitive fluorescent dyes). I obtained the voltage dependent fluorescent change from Cy5 dye by stimulating Cy3 (FRET) when Hv1/VSOP was labeled by both dyes. The kinetics of FRET efficiency was different from the non-FRET fluorescent changes during depolarization, indicating that the channel opening has two or more structural change steps.
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