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Biased Brownian stepping rotation of FoF1-ATP synthase driven by proton motive force

Nature Communications volume 4, Article number: 1631 (2013) | Download Citation

Abstract

FoF1-ATP synthase (FoF1) produces most of the ATP in cells, uniquely, by converting the proton motive force (pmf) into ATP production via mechanical rotation of the inner rotor complex. Technical difficulties have hampered direct investigation of pmf-driven rotation, which are crucial to elucidating the chemomechanical coupling mechanism of FoF1. Here we develop a novel supported membrane system for direct observation of the rotation of FoF1 driven by pmf that was formed by photolysis of caged protons. Upon photolysis, FoF1 initiated rotation in the opposite direction to that of the ATP-driven rotation. The step size of pmf-driven rotation was 120°, suggesting that the kinetic bottleneck is a catalytic event on F1 with threefold symmetry. The reaction equilibrium was slightly biased to ATP synthesis like under physiological conditions, and FoF1 showed highly stochastic behaviour, frequently making a 120° backward step. This new experimental system would be applicable to single-molecule study of other membrane proteins.

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Acknowledgements

We thank L. Yamauchi, R. Hasegawa, M. Tanigawara, J. Ki, H. Yaginuma and E. Muneyuki for critical discussion and sample preparation, and all members of Noji Laboratory for technical supports. This work was supported by Grant-in-Aid for Scientific Research No. 18074005 to H.N., No. 30540108 to R.W. and No. 23107720 to R.I. from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Affiliations

  1. Department of Applied Chemistry, School of Engineering, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-8656, Japan

    • Rikiya Watanabe
    • , Kazuhito V. Tabata
    • , Ryota Iino
    •  & Hiroyuki Noji
  2. Department of Physics, Faculty of Science and Engineering, Chuo University, Tokyo 112-8551, Japan

    • Hiroshi Ueno
  3. Department of Molecular Physiology and Biophysics, University of Fukui Faculty of Medical Sciences, Fukui 910-1193, Japan

    • Masayuki Iwamoto
    •  & Shigetoshi Oiki

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Contributions

R.W. designed and performed experiments and analysed data; K.V.T., R.I., H.U., M.I. and S.O. gave technical supports and conceptual advices; H.N. designed experiments, built whole story and wrote papers with R.W., K.V.T. and R.I.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Hiroyuki Noji.

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https://doi.org/10.1038/ncomms2631

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