Protonation-dependent stepped rotation of the F-type ATP synthase c-ring observed by single-molecule measurements

Seiga Yanagisawa, Wayne Frasch

    Research output: Contribution to journalArticle

    2 Citations (Scopus)

    Abstract

    The two opposed rotary molecular motors of the F0F1-ATP synthase work together to provide the majority of ATP in biological organisms. Rotation occurs in 120° power strokes separated by dwells when F1 synthesizes or hydrolyzes ATP. F0 and F1 complexes connect via a central rotor stalk and a peripheral stator stalk. A major unresolved question is the mechanism in which the interaction between subunit-a and rotating subunit-c–ring in the F0 motor uses the flux of H across the membrane to induce clockwise rotation against the force of counterclockwise rotation driven by the F1-ATPase. In single-molecule measurements of F0F1 embedded in lipid bilayer nanodiscs, we observed that the ability of the F0 motor to form transient dwells increases with decreasing pH. Transient dwells can halt counterclockwise rotation powered by the F1-ATPase in steps equivalent to the rotation of single c-subunits in the c-ring of F0, and can push the common axle shared by the two motors clockwise by as much as one c-subunit. Because the F0 proton half-channels that access the periplasm and the cytoplasm are exposed to the same pH, these data are consistent with the conclusion that the periplasmic half-channel is more easily protonated in a manner that halts ATPase-driven rotation by blocking ATPase-dependent proton pumping. The fit of transient dwell occurrence to the sum of three Gaussian curves suggests that the asymmetry of the three ATPase-dependent 120° power strokes imposed by the relative positions of the central and peripheral stalks affects c-subunit stepping efficiency.

    Original languageEnglish (US)
    Pages (from-to)17093-17100
    Number of pages8
    JournalJournal of Biological Chemistry
    Volume292
    Issue number41
    DOIs
    StatePublished - 2017

    Fingerprint

    Protonation
    Adenosine Triphosphate
    Molecules
    Adenosine Triphosphatases
    Proton-Translocating ATPases
    Protons
    Stroke
    Periplasm
    Lipid bilayers
    Axles
    Lipid Bilayers
    Stators
    Cytoplasm
    Rotors
    Fluxes
    Membranes

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Cite this

    Protonation-dependent stepped rotation of the F-type ATP synthase c-ring observed by single-molecule measurements. / Yanagisawa, Seiga; Frasch, Wayne.

    In: Journal of Biological Chemistry, Vol. 292, No. 41, 2017, p. 17093-17100.

    Research output: Contribution to journalArticle

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    abstract = "The two opposed rotary molecular motors of the F0F1-ATP synthase work together to provide the majority of ATP in biological organisms. Rotation occurs in 120° power strokes separated by dwells when F1 synthesizes or hydrolyzes ATP. F0 and F1 complexes connect via a central rotor stalk and a peripheral stator stalk. A major unresolved question is the mechanism in which the interaction between subunit-a and rotating subunit-c–ring in the F0 motor uses the flux of H across the membrane to induce clockwise rotation against the force of counterclockwise rotation driven by the F1-ATPase. In single-molecule measurements of F0F1 embedded in lipid bilayer nanodiscs, we observed that the ability of the F0 motor to form transient dwells increases with decreasing pH. Transient dwells can halt counterclockwise rotation powered by the F1-ATPase in steps equivalent to the rotation of single c-subunits in the c-ring of F0, and can push the common axle shared by the two motors clockwise by as much as one c-subunit. Because the F0 proton half-channels that access the periplasm and the cytoplasm are exposed to the same pH, these data are consistent with the conclusion that the periplasmic half-channel is more easily protonated in a manner that halts ATPase-driven rotation by blocking ATPase-dependent proton pumping. The fit of transient dwell occurrence to the sum of three Gaussian curves suggests that the asymmetry of the three ATPase-dependent 120° power strokes imposed by the relative positions of the central and peripheral stalks affects c-subunit stepping efficiency.",
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