Visualization of rotation of ATP synthase driven by proton motive force using single-molecule techniques and microdevices

我们试图直接观察质子动力驱动的ATP合成酶的旋转。ATP合酶被重组到支持在固体基质如NTA-琼脂糖和NTA-丙烯酰胺上的脂质双层膜中。用玻璃或聚二甲基硅氧烷制成的移液器检查支撑膜的电密封和电压分配。实现了几十兆欧姆的电阻，并用于密封含有由ATP水解驱动的旋转ATP合酶的膜。部分ATP合成酶在电压作用下随转速的变化而变化。我们需要精心的对照实验来确认这种变化是否真的反映了ATP合酶的行为。

We have tried to directly observe the rotation of ATP synthase driven by proton motive force. ATP synthase was reconstituted into the lipid bilayer membrane supported on the solid substrate such as NTA-agarose and NTA-acrylamide. Electric sealing of and voltage allocation to the supported membrane were examined using pipette made by glass or polydimethylsiloxane. Several tens of mega ohm resistance was achieved, and used to seal the membrane containing the rotating ATP synthase driven by ATP hydrolysis. Some of ATP synthase showed change in the rotation speed when voltage was applied. We need elaborate control experiments to confirm whether the change really reflects the behavior of the ATP synthase.