Coupling of Rotation and Catalysis in F1-ATPase Revealed by Single-Molecule Imaging and Manipulation

Kengo Adachi; Kazuhiro Oiwa; Takayuki Nishizaka; Shou Furuike; Hiroyuki Noji; Hiroyasu Itoh; Masasuke Yoshida; Kazuhiko Kinosita

doi:10.1016/j.cell.2007.05.020

Coupling of Rotation and Catalysis in F₁-ATPase Revealed by Single-Molecule Imaging and Manipulation

Kengo Adachi, Kazuhiro Oiwa, Takayuki Nishizaka, Shou Furuike, Hiroyuki Noji, Hiroyasu Itoh, Masasuke Yoshida, Kazuhiko Kinosita^*

^*Corresponding author for this work

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

327 Citations (Scopus)

Abstract

F₁-ATPase is a rotary molecular motor that proceeds in 120° steps, each driven by ATP hydrolysis. How the chemical reactions that occur in three catalytic sites are coupled to mechanical rotation is the central question. Here, we show by high-speed imaging of rotation in single molecules of F₁ that phosphate release drives the last 40° of the 120° step, and that the 40° rotation accompanies reduction of the affinity for phosphate. We also show, by single-molecule imaging of a fluorescent ATP analog Cy3-ATP while F₁ is forced to rotate slowly, that release of Cy3-ADP occurs at ∼240° after it is bound as Cy3-ATP at 0°. This and other results suggest that the affinity for ADP also decreases with rotation, and thus ADP release contributes part of energy for rotation. Together with previous results, the coupling scheme is now basically complete.

Original language	English
Pages (from-to)	309-321
Number of pages	13
Journal	Cell
Volume	130
Issue number	2
DOIs	https://doi.org/10.1016/j.cell.2007.05.020
Publication status	Published - 2007 Jul 27

Keywords

CHEMBIO
PROTEINS

ASJC Scopus subject areas

Cell Biology
Molecular Biology

Access to Document

10.1016/j.cell.2007.05.020

Cite this

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title = "Coupling of Rotation and Catalysis in F1-ATPase Revealed by Single-Molecule Imaging and Manipulation",

abstract = "F1-ATPase is a rotary molecular motor that proceeds in 120° steps, each driven by ATP hydrolysis. How the chemical reactions that occur in three catalytic sites are coupled to mechanical rotation is the central question. Here, we show by high-speed imaging of rotation in single molecules of F1 that phosphate release drives the last 40° of the 120° step, and that the 40° rotation accompanies reduction of the affinity for phosphate. We also show, by single-molecule imaging of a fluorescent ATP analog Cy3-ATP while F1 is forced to rotate slowly, that release of Cy3-ADP occurs at ∼240° after it is bound as Cy3-ATP at 0°. This and other results suggest that the affinity for ADP also decreases with rotation, and thus ADP release contributes part of energy for rotation. Together with previous results, the coupling scheme is now basically complete.",

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AU - Adachi, Kengo

AU - Oiwa, Kazuhiro

AU - Nishizaka, Takayuki

AU - Furuike, Shou

AU - Noji, Hiroyuki

AU - Itoh, Hiroyasu

AU - Yoshida, Masasuke

AU - Kinosita, Kazuhiko

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