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

    Research output: Contribution to journalArticle

    42 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 languageEnglish
    Article number1022
    JournalNature Communications
    Volume3
    DOIs
    Publication statusPublished - 2012

    Fingerprint

    Proton-Translocating ATPases
    adenosine triphosphate
    Adenosine Triphosphate
    synthesis
    adenosine diphosphate
    Adenosine Diphosphate
    affinity
    Magnets
    rotors
    hydrolysis
    Hydrolysis
    Catalytic Domain
    phosphates
    magnets
    Rotors
    Phosphates

    ASJC Scopus subject areas

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

    Cite this

    Controlled rotation of the F 1 -ATPase reveals differential and continuous binding changes for ATP synthesis. / Adachi, Kengo; Oiwa, Kazuhiro; Yoshida, Masasuke; Nishizaka, Takayuki; Kinosita, Kazuhiko.

    In: Nature Communications, Vol. 3, 1022, 2012.

    Research output: Contribution to journalArticle

    Adachi, Kengo ; Oiwa, Kazuhiro ; Yoshida, Masasuke ; Nishizaka, Takayuki ; Kinosita, Kazuhiko. / Controlled rotation of the F 1 -ATPase reveals differential and continuous binding changes for ATP synthesis. In: Nature Communications. 2012 ; Vol. 3.
    @article{a55102e4776447d18f6bb178295aa73f,
    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.",
    author = "Kengo Adachi and Kazuhiro Oiwa and Masasuke Yoshida and Takayuki Nishizaka and Kazuhiko Kinosita",
    year = "2012",
    doi = "10.1038/ncomms2026",
    language = "English",
    volume = "3",
    journal = "Nature Communications",
    issn = "2041-1723",
    publisher = "Nature Publishing Group",

    }

    TY - JOUR

    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.

    UR - http://www.scopus.com/inward/record.url?scp=84866110008&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=84866110008&partnerID=8YFLogxK

    U2 - 10.1038/ncomms2026

    DO - 10.1038/ncomms2026

    M3 - Article

    C2 - 22929779

    AN - SCOPUS:84866110008

    VL - 3

    JO - Nature Communications

    JF - Nature Communications

    SN - 2041-1723

    M1 - 1022

    ER -