Temperature dependence of the rotation and hydrolysis activities of F 1-ATPase

Shou Furuike, Kengo Adachi, Naoyoshi Sakaki, Rieko Shimo-Kon, Hiroyasu Itoh, Eiro Muneyuki, Masasuke Yoshida, Kazuhiko Kinosita

    Research output: Contribution to journalArticle

    28 Citations (Scopus)

    Abstract

    F1-ATPase, a water-soluble portion of the enzyme ATP synthase, is a rotary molecular motor driven by ATP hydrolysis. To learn how the kinetics of rotation are regulated, we have investigated the rotational characteristics of a thermophilic F1-ATPase over the temperature range 4-50°C by attaching a polystyrene bead (or bead duplex) to the rotor subunit and observing its rotation under a microscope. The apparent rate of ATP binding estimated at low ATP concentrations increased from 1.2 × 106 M-1 s-1 at 4°C to 4.3 × 107 M-1 s -1 at 40°C, whereas the torque estimated at 2 mM ATP remained around 40 pN·nm over 4-50°C. The rotation was stepwise at 4°C, even at the saturating ATP concentration of 2 mM, indicating the presence of a hitherto unresolved rate-limiting reaction that occurs at ATP-waiting angles. We also measured the ATP hydrolysis activity in bulk solution at 4-65°C. F1-ATPase tends to be inactivated by binding ADP tightly. Both the inactivation and reactivation rates were found to rise sharply with temperature, and above 30°C, equilibrium between the active and inactive forms was reached within 2 s, the majority being inactive. Rapid inactivation at high temperatures is consistent with the physiological role of this enzyme, ATP synthesis, in the thermophile.

    Original languageEnglish
    Pages (from-to)761-770
    Number of pages10
    JournalBiophysical Journal
    Volume95
    Issue number2
    DOIs
    Publication statusPublished - 2008 Jul 15

    Fingerprint

    Proton-Translocating ATPases
    Hydrolysis
    Adenosine Triphosphate
    Temperature
    Polystyrenes
    Torque
    Enzymes
    Adenosine Diphosphate
    Water

    ASJC Scopus subject areas

    • Biophysics

    Cite this

    Furuike, S., Adachi, K., Sakaki, N., Shimo-Kon, R., Itoh, H., Muneyuki, E., ... Kinosita, K. (2008). Temperature dependence of the rotation and hydrolysis activities of F 1-ATPase. Biophysical Journal, 95(2), 761-770. https://doi.org/10.1529/biophysj.107.123307

    Temperature dependence of the rotation and hydrolysis activities of F 1-ATPase. / Furuike, Shou; Adachi, Kengo; Sakaki, Naoyoshi; Shimo-Kon, Rieko; Itoh, Hiroyasu; Muneyuki, Eiro; Yoshida, Masasuke; Kinosita, Kazuhiko.

    In: Biophysical Journal, Vol. 95, No. 2, 15.07.2008, p. 761-770.

    Research output: Contribution to journalArticle

    Furuike, S, Adachi, K, Sakaki, N, Shimo-Kon, R, Itoh, H, Muneyuki, E, Yoshida, M & Kinosita, K 2008, 'Temperature dependence of the rotation and hydrolysis activities of F 1-ATPase', Biophysical Journal, vol. 95, no. 2, pp. 761-770. https://doi.org/10.1529/biophysj.107.123307
    Furuike S, Adachi K, Sakaki N, Shimo-Kon R, Itoh H, Muneyuki E et al. Temperature dependence of the rotation and hydrolysis activities of F 1-ATPase. Biophysical Journal. 2008 Jul 15;95(2):761-770. https://doi.org/10.1529/biophysj.107.123307
    Furuike, Shou ; Adachi, Kengo ; Sakaki, Naoyoshi ; Shimo-Kon, Rieko ; Itoh, Hiroyasu ; Muneyuki, Eiro ; Yoshida, Masasuke ; Kinosita, Kazuhiko. / Temperature dependence of the rotation and hydrolysis activities of F 1-ATPase. In: Biophysical Journal. 2008 ; Vol. 95, No. 2. pp. 761-770.
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