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The ATP-waiting conformation of rotating F1-ATPase revealed by single-pair fluorescence resonance energy transfer
- Ryohei Yasuda*,†,
- Tomoko Masaike‡,§,
- Kengo Adachi¶,
- Hiroyuki Noji∥,
- Hiroyasu Itoh**, and
- Kazuhiko Kinosita, Jr.¶
- *Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724; ‡ATP System Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, 5800-3 Nagatsuta, Yokohama 226-0026, Japan; §Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama 226-8503, Japan; ¶Center for Integrative Bioscience, Okazaki National Research Institutes, Higashiyama 5-1, Myodaiji, Okazaki 444-8585, Japan; ∥Precursory Research for Embryonic Science and Technology, Japan Science and Technology Corporation, Institute of Industrial Science, University of Tokyo, 4-6-1 Komaba, Tokyo 153-8505, Japan; and **Hamamatsu Photonics KK, Tokodai, Tsukuba 300-2635, Japan
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Edited by Paul D. Boyer, University of California, Los Angeles, CA, and approved June 16, 2003 (received for review December 23, 2002)
Abstract
F1-ATPase is an ATP-driven rotary motor in which a rod-shaped γ subunit rotates inside a cylinder made of α3β3 subunits. To elucidate the conformations of rotating F1, we measured fluorescence resonance energy transfer (FRET) between a donor on one of the three βs and an acceptor on γ in single F1 molecules. The yield of FRET changed stepwise at low ATP concentrations, reflecting the stepwise rotation of γ. In the ATP-waiting state, the FRET yields indicated a γ position ≈40° counterclockwise (= direction of rotation) from that in the crystal structures of mitochondrial F1, suggesting that the crystal structures mimic a metastable state before product release.
Footnotes
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↵ † To whom correspondence should be addressed. E-mail: yasuda{at}cshl.org.
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This paper was submitted directly (Track II) to the PNAS office.
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Abbreviations: FRET, fluorescence resonance energy transfer; MF1, mitochondrial F1; DCCD, dicyclohexylcarbodiimide.
- Copyright © 2003, The National Academy of Sciences
