The anlaysis of the cooperative gating of voltage-gated proton channel

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K18516
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biophysics
• Research Institution: Osaka University
• Principal Investigator: Kawanabe Akira 大阪大学, 医学系研究科, 特任助教(常勤) (10707128)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: 電位依存性タンパク質 / イオンチャネル / 電位依存性プロトンチャネル

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.

Free Research Field

生物物理学、生理学