Mechanically driven proton conduction in single δ-free F₀F₁-ATPase

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• 作者：Liu Xiaolong ; Zhang Xiaoai ; Cui Yuanbo ; Yue Jiachang ; Luo Zhiyong and Jiang Peidong
• 关键词：F₀F₁-ATPase ; Mechanically driven proton flux ; Chromatophore
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2006
• 期刊代码：159_0006291x
• 类别：bio
• 出版时间：1 September 2006
• 卷：347
• 期：3
• 页码：752-757
• 文件大小：369 K

摘要

In order to observe mechanically driven proton flux in F₀F₁-ATPase coupled with artificial driven rotation on F₁ simultaneously, a double channel observation system was established. An artificial δ-free F₀F₁-ATPase was constructed with α₃, β₃, ε, γ, and c_n subunits as rotator and a, b₂ as stator. The chromatophore was immobilized on the glass surface through biotin–streptavidin–biotin system, and the magnetic bead was attached to the β subunit of δ-free F₀F₁-ATPase. The mechanically driven proton flux was indicated by the fluorescence intensity change of fluorescein reference standard (F1300) and recorded by a cooled digital CCD camera. The mechanochemical coupling stoichiometry between F₀ and F₁ is about 4.15 ± 0.2H⁺/rev when the magnetic field rotated at 0.33 Hz (rps).