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Chemomechanical coupling in F1-ATPase revealed by simultaneous observation of nucleotide kinetics and rotation

Nature Structural & Molecular Biology volume 11, pages 142148 (2004) | Download Citation

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Abstract

F1-ATPase is a rotary molecular motor in which unidirectional rotation of the central γ subunit is powered by ATP hydrolysis in three catalytic sites arranged 120° apart around γ. To study how hydrolysis reactions produce mechanical rotation, we observed rotation under an optical microscope to see which of the three sites bound and released a fluorescent ATP analog. Assuming that the analog mimics authentic ATP, the following scheme emerges: (i) in the ATP-waiting state, one site, dictated by the orientation of γ, is empty, whereas the other two bind a nucleotide; (ii) ATP binding to the empty site drives an 80° rotation of γ; (iii) this triggers a reaction(s), hydrolysis and/or phosphate release, but not ADP release in the site that bound ATP one step earlier; (iv) completion of this reaction induces further 40° rotation.

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Acknowledgements

We thank R. Yasuda, K. Adachi and H. Itoh for technical assistance and critical discussion; R. Nakamori and Y. Funamoto for technical assistance; D.R. Trentham for initiation of our collaboration; T. Masaike, B. Brenner and H. Kojima for critical discussion; M. Shio, K. Abe and I. Sase for the microscope techniques; and M. Uno and H. Umezawa for management of laboratories and collaboration. This work was supported in part by grants-in-aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and by a Core Research for Evolutional Science and Technology (CREST) grant.

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Author notes

    • Takayuki Nishizaka

    Present address: Department of Physics, Gakushuin University, Mejiro 1-5-1, Toshima-ku, Tokyo 171-8588, Japan.

Affiliations

  1. Kansai Advanced Research Center, Protein Biophysics Group, Iwaoka 588-2, Nishi-ku, Kobe 651-2492, Japan.

    • Takayuki Nishizaka
    • , Kazuhiro Oiwa
    •  & Shigeki Kimura
  2. Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency, Honcho 4-1-8, Kawaguchi, Saitama 332-0012, Japan.

    • Takayuki Nishizaka
  3. Institute of Industrial Science, University of Tokyo, Tokyo 153-8505, Japan.

    • Hiroyuki Noji
  4. Chemical Resources Laboratory, Tokyo Institute of Technology, Yokohama 226-8503, Japan.

    • Eiro Muneyuki
    •  & Masasuke Yoshida
  5. ATP system, Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Agency, Yokohama 226-0026, Japan.

    • Masasuke Yoshida
  6. Center for Integrative Bioscience, Okazaki National Research Institutes, Myodaiji, Okazaki 444-8585, Japan.

    • Kazuhiko Kinosita Jr

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The authors declare no competing financial interests.

Corresponding author

Correspondence to Takayuki Nishizaka.

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DOI

https://doi.org/10.1038/nsmb721

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