Resolving stepping rotation in Thermus thermophilus H+-ATPase/ synthase with an essentially drag-free probe

Shou Furuike; Masahiro Nakano; Kengo Adachi; Hiroyuki Noji; Kazuhiko Kinosita; Ken Yokoyama

doi:10.1038/ncomms1215

Resolving stepping rotation in Thermus thermophilus H⁺-ATPase/ synthase with an essentially drag-free probe

Shou Furuike, Masahiro Nakano, Kengo Adachi, Hiroyuki Noji, Kazuhiko Kinosita^*, Ken Yokoyama

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

45 Citations (Scopus)

Abstract

Vacuole-type ATPases (V_oV₁) and F_oF ₁ ATP synthases couple ATP hydrolysis/synthesis in the soluble V ₁ or F₁ portion with proton (or Na⁺) flow in the membrane-embedded V_o or F_o portion through rotation of one common shaft. Here we show at submillisecond resolutions the ATP-driven rotation of isolated V₁ and the whole V_oV₁ 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. V _oV₁ exhibited 12 dwell positions per revolution, consistent with the 12-fold symmetry of the V_o rotor in T. thermophilus. Unlike F 1 that undergoes 80 °-40 ° substepping, chemo-mechanical checkpoints in isolated V₁ are all at the ATP-waiting position, and V_o adds further bumps through stator-rotor interactions outside and remote from V₁.

Original language	English
Article number	233
Journal	Nature Communications
Volume	2
Issue number	1
DOIs	https://doi.org/10.1038/ncomms1215
Publication status	Published - 2011

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Chemistry(all)
Physics and Astronomy(all)

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10.1038/ncomms1215

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title = "Resolving stepping rotation in Thermus thermophilus H+-ATPase/ synthase with an essentially drag-free probe",

abstract = "Vacuole-type ATPases (VoV1) and FoF 1 ATP synthases couple ATP hydrolysis/synthesis in the soluble V 1 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. V oV1 exhibited 12 dwell positions per revolution, consistent with the 12-fold symmetry of the Vo rotor in T. thermophilus. Unlike F 1 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.",

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AU - Kinosita, Kazuhiko

AU - Yokoyama, Ken

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N2 - Vacuole-type ATPases (VoV1) and FoF 1 ATP synthases couple ATP hydrolysis/synthesis in the soluble V 1 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. V oV1 exhibited 12 dwell positions per revolution, consistent with the 12-fold symmetry of the Vo rotor in T. thermophilus. Unlike F 1 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.

AB - Vacuole-type ATPases (VoV1) and FoF 1 ATP synthases couple ATP hydrolysis/synthesis in the soluble V 1 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. V oV1 exhibited 12 dwell positions per revolution, consistent with the 12-fold symmetry of the Vo rotor in T. thermophilus. Unlike F 1 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.

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Resolving stepping rotation in Thermus thermophilus H⁺-ATPase/ synthase with an essentially drag-free probe

Abstract

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