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

    Research output: Contribution to journalArticle

    281 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 languageEnglish
    Pages (from-to)309-321
    Number of pages13
    JournalCell
    Volume130
    Issue number2
    DOIs
    Publication statusPublished - 2007 Jul 27

    Keywords

    • CHEMBIO
    • PROTEINS

    ASJC Scopus subject areas

    • Cell Biology
    • Molecular Biology

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