Resolving stepping rotation in Thermus thermophilus H(+)-ATPase/synthase with an essentially drag-free probe.
Resolving stepping rotation in Thermus thermophilus H(+)-ATPase/synthase with an essentially drag-free probe.
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用基本无阻力的探针在热嗜热H（+）ATPase/Synthase中解析垫脚旋转。
DOI：
10.1038/ncomms1215
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发表时间：
2011
影响因子：
16.6
通讯作者：
Yokoyama, Ken
中科院分区：
文献类型：
--
作者：
Furuike, Shou;Nakano, Masahiro;Adachi, Kengo;Noji, Hiroyuki;Kinosita, Kazuhiko, Jr.;Yokoyama, Ken
胆甾醇型ATP酶（VoV 1）和FoF 1 ATP酶通过一个共同轴的旋转将可溶性V1或F1部分中的ATP水解/合成与膜包埋的Vo或Fo部分中的质子（或Na+）流偶联。在这里，我们显示在亚毫秒分辨率的ATP驱动的旋转孤立的V1和整个VoV 1从嗜热栖热菌，通过附加一个40纳米的金珠的粘性阻力几乎可以忽略不计。V1产生120°台阶，与三个催化位点的存在相称。步骤之间的停留涉及至少两个事件，而不是ATP结合，一个可能是ATP水解。VoV 1每转显示12个停留位置，与T中Vo转子的12重对称性一致。嗜热菌与经历80°-40°细分的F1不同，隔离V1中的化学机械检查点都在ATP等待位置，并且Vo通过V1外部和远离V1的定子-转子相互作用增加了进一步的颠簸。旋转ATP酶FoF 1和VoV 1将ATP水解与质子流偶联。Furuike等人以亚毫秒分辨率观察V1和VoV 1中ATP驱动的旋转，发现V1中的限速反应都发生在相同的角度，Vo中的定子-转子相互作用引入了额外的检查点。
Vacuole-type ATPases (VoV1) and FoF1 ATP synthases couple ATP hydrolysis/synthesis in the soluble V1 or F1 portion with proton (or Na+) flow in the membrane-embedded Vo or Fo portion through rotation of one common shaft. Here we show at submillisecond resolutions the ATP-driven rotation of isolated V1 and the whole VoV1 from Thermus thermophilus, by attaching a 40-nm gold bead for which viscous drag is almost negligible. V1 made 120° steps, commensurate with the presence of three catalytic sites. Dwells between the steps involved at least two events other than ATP binding, one likely to be ATP hydrolysis. VoV1 exhibited 12 dwell positions per revolution, consistent with the 12-fold symmetry of the Vo rotor in T. thermophilus. Unlike F1 that undergoes 80°–40° substepping, chemo-mechanical checkpoints in isolated V1 are all at the ATP-waiting position, and Vo adds further bumps through stator–rotor interactions outside and remote from V1. Rotary ATPases FoF1 and VoV1 couple ATP hydrolysis with proton flow. Furuike et al. observe ATP-driven rotation in V1 and VoV1, at submillisecond resolution, and find that rate-limiting reactions in V1 all occur at the same angle, and stator–rotor interactions in Vo introduce additional checkpoints.
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DOI：
10.1073/pnas.031564198
发表时间：
2001-01-30
影响因子：
11.1
作者：
Tsunoda, SP;Aggeler, R;Capaldi, RA
通讯作者：
Capaldi, RA
影响因子：
16.8
作者：
Diez, M;Zimmermann, B;Gräber, P
通讯作者：
Gräber, P
影响因子：
3
作者：
Yokoyama, K;Imamura, H
通讯作者：
Imamura, H
DOI：
10.1073/pnas.0407857102
发表时间：
2005-02-01
影响因子：
11.1
作者：
Ueno, H;Suzuki, T;Yoshida, M
通讯作者：
Yoshida, M
DOI：
10.1073/pnas.0406486102
发表时间：
2005-03-22
影响因子：
11.1
作者：
Hirono-Hara, Y;Ishuzuka, K;Noji, H
通讯作者：
Noji, H