Crio eletromicroscopia da estrutura da rotativa intacta da H + -ATPase / sintase do Thermus thermophilus: mero acaso, fortuita necessidade ou design inteligente?

sexta-feira, fevereiro 02, 2018

Cryo EM structure of intact rotary H+-ATPase/synthase from Thermus thermophilus

Atsuko Nakanishi, Jun-ichi Kishikawa, Masatada Tamakoshi, Kaoru Mitsuoka & Ken Yokoyama

Nature Communications volume 9, Article number: 89 (2018)

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Bioenergetics Cryoelectron microscopy Enzyme mechanisms

Received: 10 August 2017 Accepted: 04 December 2017

Published online: 08 January 2018


Proton translocating rotary ATPases couple ATP hydrolysis/synthesis, which occurs in the soluble domain, with proton flow through the membrane domain via a rotation of the common central rotor complex against the surrounding peripheral stator apparatus. Here, we present a large data set of single particle cryo-electron micrograph images of the V/A type H+-rotary ATPase from the bacterium Thermus thermophilus, enabling the identification of three rotational states based on the orientation of the rotor subunit. Using masked refinement and classification with signal subtractions, we obtain homogeneous reconstructions for the whole complexes and soluble V1 domains. These reconstructions are of higher resolution than any EM map of intact rotary ATPase reported previously, providing a detailed molecular basis for how the rotary ATPase maintains structural integrity of the peripheral stator apparatus, and confirming the existence of a clear proton translocation path from both sides of the membrane.


We thank Dr Bernadette Byrne for critical reading of our manuscript, and members of Yokoyama Lab for help and technical assistance. This work was supported by Grants-in-Aid from “Nanotechnology Platform” of the Ministry of Education, Culture, Sports, Science and Technology (MEXT) to K.M. (Project No. 12024046) and Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) to K.Y. (Project No. 17H03648), and J.K. (Project No. 16K21472).

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Author notes

Atsuko Nakanishi and Jun-ichi Kishikawa contributed equally to this work.


Department of Molecular Biosciences, Kyoto Sangyo University, Motoyama Kamigamo, Kita-ku, Kyoto, 603-8555, Japan

Atsuko Nakanishi, Jun-ichi Kishikawa & Ken Yokoyama

Department of Molecular Biology, Tokyo University of Pharmacy and Life Sciences, Horinouchi, Hachioji, Tokyo, 192-0392, Japan

Masatada Tamakoshi

Research Center for Ultra-High Voltage Electron Microscopy, Osaka University, 7-1, Mihogaoka, Ibaraki, Osaka, 567-0047, Japan

Kaoru Mitsuoka


A.N., J.K. designed, performed, and analyzed the experiments. A.N., J.K., K.M. analyzed the data and contributed to the preparation of the figures. M.T. constructed vectors for expression of mutant proteins. K.M. gave technical support and conceptual advice. K.Y. designed and supervised the experiments and wrote the manuscript. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Kaoru Mitsuoka or Ken Yokoyama.