高分解顕微鏡観察を主としたカリウムチャネルKcsAの柔軟な構造と機能の相関解明

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Author: 内橋, 貴之ほか

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Publication date: 2016-06-13

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Note (General)：: 金沢大学理工研究域数物科学系電子線リソグラフィーと収束イオンビーム(FIB)を用いて固体基板に直径100nmのナノ開口アレイを製作し、ナノ開口を脂質二重膜で封止することができた。また、紫膜でナノ開口を封止することで、中空膜でバクテリオロドプシン２次元結晶を高速AFMで高解像観察することに成功した。さ...

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Material Type: 文書・図像類
Title: 高分解顕微鏡観察を主としたカリウムチャネルKcsAの柔軟な構造と機能の相関解明
Author/Editor: 内橋, 貴之
Uchihashi, Takayuki
Author Heading: 内橋, 貴之
Uchihashi, Takayuki
Publication Date: 2016-06-13
Publication Date (W3CDTF): 2016-06-13
Alternative Title: Development of Non-Supported Membrane over Nanowells for Visualization of Functional Dynamics of Membrane Proteins with High-Speed AFM
Periodical title: 平成27(2015)年度科学研究費補助金挑戦的萌芽研究研究成果報告書 = 2015 Fiscal Year Final Research Report
No. or year of volume/issue: 2014-04-01 - 2016-03-31
Volume: 2014-04-01 - 2016-03-31
Pages: 4p.-
Text Language Code: jpn
Subject Heading: 一分子計測
膜タンパク質
高速原子間力顕微鏡
Target Audience: 一般
Note (General): 金沢大学理工研究域数物科学系
電子線リソグラフィーと収束イオンビーム(FIB)を用いて固体基板に直径100nmのナノ開口アレイを製作し、ナノ開口を脂質二重膜で封止することができた。また、紫膜でナノ開口を封止することで、中空膜でバクテリオロドプシン２次元結晶を高速AFMで高解像観察することに成功した。さらに、直径100nmのガラスあるいはポリスチレンビーズを鋳型としてシリコーンゴム(PDMS)に多数のナノ開口を簡便に製作することにも成功した。基板支持膜に再構成されたイオンチャネルKcsAの四量体と拡散ダイナミクスを高速AFM観察することにも成功した。今後、KcsAを中空膜に再構成し機能動態の高速AFM観察を行っていく。
We prepared nano-pore arrays with a diameter of 100nm on glass and silicon substrate using electron-beam lithography and focused ion beam. Also we succeeded in sealing the nano pores with lipid bilayers. Purple membranes in which two dimensional crystals of bacteriorhodopsin are embedded were used as a test sample and the individual bacteriorhodopsin molecules were resolved on the nano pore with high-speed AFM. We also developed a method to conveniently prepare the nano pores on a silicone elastomer using small beads with a diameter of 100nm as a template. We observed ion-channel membrane protein KcsA on a supported lipid bilayer and visualized individual KcsA tetramers and theirs diffusion dynamics in the membrane. As the next step, we will reconstitute KcsA molecules into the membrane on the nano pore and reveal structural dynamics of KcsA.
研究課題/領域番号:26600052, 研究期間(年度):2014-04-01 - 2016-03-31
DOI: 10.24517/00056742
Access Restrictions: 限定公開
Related Material: https://kaken.nii.ac.jp/search/?qm=30326300
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-26600052/
https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-26600052/26600052seika/
Data Provider (Database): 国立情報学研究所 : 学術機関リポジトリデータベース（IRDB）（機関リポジトリ）
https://irdb.nii.ac.jp
Original Data Provider (Database): 金沢大学 : 金沢大学学術情報リポジトリKURA