Controlled rotation of the F 1 -ATPase reveals differential and continuous binding changes for ATP synthesis

Kengo Adachi; Kazuhiro Oiwa; Masasuke Yoshida; Takayuki Nishizaka; Kazuhiko Kinosita

doi:10.1038/ncomms2026

Controlled rotation of the F 1 -ATPase reveals differential and continuous binding changes for ATP synthesis

Kengo Adachi^*, Kazuhiro Oiwa, Masasuke Yoshida, Takayuki Nishizaka, Kazuhiko Kinosita

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

57 Citations (Scopus)

Abstract

F 1 -ATPase is an ATP-driven rotary molecular motor that synthesizes ATP when rotated in reverse. To elucidate the mechanism of ATP synthesis, we imaged binding and release of fluorescently labelled ADP and ATP while rotating the motor in either direction by magnets. Here we report the binding and release rates for each of the three catalytic sites for 360°C of the rotary angle. We show that the rates do not significantly depend on the rotary direction, indicating ATP synthesis by direct reversal of the hydrolysis-driven rotation. ADP and ATP are discriminated in angle-dependent binding, but not in release. Phosphate blocks ATP binding at angles where ADP binding is essential for ATP synthesis. In synthesis rotation, the affinity for ADP increases by >10 4, followed by a shift to high ATP affinity, and finally the affinity for ATP decreases by >10 4. All these angular changes are gradual, implicating tight coupling between the rotor angle and site affinities.

Original language	English
Article number	1022
Journal	Nature Communications
Volume	3
DOIs	https://doi.org/10.1038/ncomms2026
Publication status	Published - 2012

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Chemistry(all)
Physics and Astronomy(all)

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title = "Controlled rotation of the F 1 -ATPase reveals differential and continuous binding changes for ATP synthesis",

abstract = "F 1 -ATPase is an ATP-driven rotary molecular motor that synthesizes ATP when rotated in reverse. To elucidate the mechanism of ATP synthesis, we imaged binding and release of fluorescently labelled ADP and ATP while rotating the motor in either direction by magnets. Here we report the binding and release rates for each of the three catalytic sites for 360°C of the rotary angle. We show that the rates do not significantly depend on the rotary direction, indicating ATP synthesis by direct reversal of the hydrolysis-driven rotation. ADP and ATP are discriminated in angle-dependent binding, but not in release. Phosphate blocks ATP binding at angles where ADP binding is essential for ATP synthesis. In synthesis rotation, the affinity for ADP increases by >10 4, followed by a shift to high ATP affinity, and finally the affinity for ATP decreases by >10 4. All these angular changes are gradual, implicating tight coupling between the rotor angle and site affinities.",

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T1 - Controlled rotation of the F 1 -ATPase reveals differential and continuous binding changes for ATP synthesis

AU - Adachi, Kengo

AU - Oiwa, Kazuhiro

AU - Yoshida, Masasuke

AU - Nishizaka, Takayuki

AU - Kinosita, Kazuhiko

PY - 2012

Y1 - 2012

N2 - F 1 -ATPase is an ATP-driven rotary molecular motor that synthesizes ATP when rotated in reverse. To elucidate the mechanism of ATP synthesis, we imaged binding and release of fluorescently labelled ADP and ATP while rotating the motor in either direction by magnets. Here we report the binding and release rates for each of the three catalytic sites for 360°C of the rotary angle. We show that the rates do not significantly depend on the rotary direction, indicating ATP synthesis by direct reversal of the hydrolysis-driven rotation. ADP and ATP are discriminated in angle-dependent binding, but not in release. Phosphate blocks ATP binding at angles where ADP binding is essential for ATP synthesis. In synthesis rotation, the affinity for ADP increases by >10 4, followed by a shift to high ATP affinity, and finally the affinity for ATP decreases by >10 4. All these angular changes are gradual, implicating tight coupling between the rotor angle and site affinities.

AB - F 1 -ATPase is an ATP-driven rotary molecular motor that synthesizes ATP when rotated in reverse. To elucidate the mechanism of ATP synthesis, we imaged binding and release of fluorescently labelled ADP and ATP while rotating the motor in either direction by magnets. Here we report the binding and release rates for each of the three catalytic sites for 360°C of the rotary angle. We show that the rates do not significantly depend on the rotary direction, indicating ATP synthesis by direct reversal of the hydrolysis-driven rotation. ADP and ATP are discriminated in angle-dependent binding, but not in release. Phosphate blocks ATP binding at angles where ADP binding is essential for ATP synthesis. In synthesis rotation, the affinity for ADP increases by >10 4, followed by a shift to high ATP affinity, and finally the affinity for ATP decreases by >10 4. All these angular changes are gradual, implicating tight coupling between the rotor angle and site affinities.

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Controlled rotation of the F 1 -ATPase reveals differential and continuous binding changes for ATP synthesis

Abstract

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