Activation and stiffness of the inhibited states of F1-ATPase probed by single-molecule manipulation

Ei Ichiro Saita, Ryota Iino, Toshiharu Suzuki, Boris A. Feniouk, Kazuhiko Kinosita, Masasuke Yoshida

    Research output: Contribution to journalArticle

    26 Citations (Scopus)

    Abstract

    F1-ATPase (F1), a soluble portion of F oF1-ATP synthase (FoF1), is an ATP-driven motor in which γε subunits rotate in the α3β3 cylinder. Activity of F1 and FoF1 from Bacillus PS3 is attenuated by the ε subunit in an inhibitory extended form. In this study we observed ATP-dependent transition of ε in single F1 molecules from extended form to hairpin form by fluorescence resonance energy transfer. The results justify the previous bulk experiments and ensure that fraction of F1 with hairpin ε directly determines the fraction of active F1 at any ATP concentration. Next, mechanical activation and stiffness of ε-inhibited F1 were examined by the forced rotation of magnetic beads attached to γ. Compared with ADP inhibition, which is another manner of inhibition, rotation by a larger angle was required for the activation from ε inhibition when the beads were forced to rotate to ATP hydrolysis direction, and more torque was required to reach the same rotation angle when beads were forced to rotate to ATP synthesis direction. The results imply that if FoF1 is resting in the ε-inhibited state, Fo motor must transmit to γ a torque larger than expected from thermodynamic equilibrium to initiate ATP synthesis.

    Original languageEnglish
    Pages (from-to)11411-11417
    Number of pages7
    JournalJournal of Biological Chemistry
    Volume285
    Issue number15
    DOIs
    Publication statusPublished - 2010 Apr 9

    Fingerprint

    Proton-Translocating ATPases
    Adenosine Triphosphate
    Chemical activation
    Stiffness
    Molecules
    Torque
    Fluorescence Resonance Energy Transfer
    Bacilli
    Engine cylinders
    Thermodynamics
    Adenosine Diphosphate
    Bacillus
    Hydrolysis

    ASJC Scopus subject areas

    • Biochemistry
    • Cell Biology
    • Molecular Biology

    Cite this

    Saita, E. I., Iino, R., Suzuki, T., Feniouk, B. A., Kinosita, K., & Yoshida, M. (2010). Activation and stiffness of the inhibited states of F1-ATPase probed by single-molecule manipulation. Journal of Biological Chemistry, 285(15), 11411-11417. https://doi.org/10.1074/jbc.M109.099143

    Activation and stiffness of the inhibited states of F1-ATPase probed by single-molecule manipulation. / Saita, Ei Ichiro; Iino, Ryota; Suzuki, Toshiharu; Feniouk, Boris A.; Kinosita, Kazuhiko; Yoshida, Masasuke.

    In: Journal of Biological Chemistry, Vol. 285, No. 15, 09.04.2010, p. 11411-11417.

    Research output: Contribution to journalArticle

    Saita, EI, Iino, R, Suzuki, T, Feniouk, BA, Kinosita, K & Yoshida, M 2010, 'Activation and stiffness of the inhibited states of F1-ATPase probed by single-molecule manipulation', Journal of Biological Chemistry, vol. 285, no. 15, pp. 11411-11417. https://doi.org/10.1074/jbc.M109.099143
    Saita, Ei Ichiro ; Iino, Ryota ; Suzuki, Toshiharu ; Feniouk, Boris A. ; Kinosita, Kazuhiko ; Yoshida, Masasuke. / Activation and stiffness of the inhibited states of F1-ATPase probed by single-molecule manipulation. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 15. pp. 11411-11417.
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