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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Evidence for rotation of V1-ATPase.
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Evidence for rotation of V1-ATPase.

机译：V1-ATPase旋转的证据。

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摘要

V(o)V(1)-ATPase is responsible for acidification of eukaryotic intracellular compartments and ATP synthesis of Archaea and some eubacteria. From the similarity to F(o)F(1)-ATP synthase, V(o)V(1)-ATPase has been assumed to be a rotary motor, but to date there are no experimental data to support this. Here we visualized the rotation of single molecules of V(1)-ATPase, a catalytic subcomplex of V(o)V(1)-ATPase. V(1)-ATPase from Thermus thermophilus was immobilized onto a glass surface, and a bead was attached to the D or F subunit through the biotin-streptavidin linkage. In both cases we observed ATP-dependent rotations of beads, the direction of which was always counterclockwise viewed from the membrane side. Given that three ATP molecules are hydrolyzed per one revolution, rates of rotation agree consistently with rates of ATP hydrolysis at saturating ATP concentrations. This study provides experimental evidence that V(o)V(1)-ATPase is a rotary motor and that both D and F subunits constitute a rotor shaft.
机译：V（o）V（1）-ATPase负责酸化真核细胞内区室和古细菌和某些真细菌的ATP合成。由于与F（o）F（1）-ATP合酶的相似性，已假定V（o）V（1）-ATPase是旋转马达，但是迄今为止，尚无实验数据来支持这一点。在这里，我们可视化V（1）-ATPase，V（o）V（1）-ATPase的催化亚复合物的单分子的旋转。将来自嗜热栖热菌的V（1）-ATPase固定在玻璃表面上，并将珠子通过生物素-链霉亲和素键连接至D或F亚基。在这两种情况下，我们都观察到了珠子的ATP依赖性旋转，从薄膜侧看，其方向始终是逆时针方向。假设每转一圈水解了三个ATP分子，则旋转速度与饱和ATP浓度下的ATP水解速度一致。这项研究提供了实验证据，表明V（o）V（1）-ATPase是一种旋转电机，D和F亚基均构成转子轴。

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