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Article
Nature Structural & Molecular Biology  11, 142 - 148 (2004)
Published online: 18 January 2004; | doi:10.1038/nsmb721

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

Takayuki Nishizaka1, 2, 7, Kazuhiro Oiwa1, Hiroyuki Noji3, Shigeki Kimura1, Eiro Muneyuki4, Masasuke Yoshida4, 5 & Kazuhiko Kinosita Jr6

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

2  Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency, Honcho 4-1-8, Kawaguchi, Saitama 332-0012, Japan.

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

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

5  ATP system, Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Agency, Yokohama 226-0026, Japan.

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

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

Correspondence should be addressed to Takayuki Nishizaka takayuki.nishizaka@gakushuin.ac.jp
F1-ATPase is a rotary molecular motor in which unidirectional rotation of the central bold gamma subunit is powered by ATP hydrolysis in three catalytic sites arranged 120° apart around bold gamma. 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 bold gamma, is empty, whereas the other two bind a nucleotide; (ii) ATP binding to the empty site drives an approx80° rotation of bold gamma; (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 approx40° rotation.

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Nature Structural & Molecular Biology
ISSN: 1545-9993
EISSN: 1545-9985
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