Torque generation and utilization in motor enzyme F 0F 1-ATP synthase: Half-torque F 1 with short-sized pushrod helix and reduced ATP synthesis by half-torque F 0F 1

Eiji Usukura, Toshiharu Suzuki, Shou Furuike, Naoki Soga, Ei Ichiro Saita, Toru Hisabori, Kazuhiko Kinosita, Masasuke Yoshida

    Research output: Contribution to journalArticle

    29 Citations (Scopus)

    Abstract

    ATP synthase (F 0F 1) is made of two motors, a proton-driven motor (F 0) and an ATP-driven motor (F 1), connected by a common rotary shaft, and catalyzes proton flow-driven ATP synthesis and ATP-driven proton pumping. In F 1, the central γ subunit rotates inside the α 3β 3 ring. Here we report structural features of F 1 responsible for torque generation and the catalytic ability of the low-torque F 0F 1. (i) Deletion of one or two turns in the α-helix in the C-terminal domain of catalytic β subunit at the rotor/stator contact region generates mutant F 1s, termed F 1(1/2)s, that rotate with about half of the normal torque. This helix would support the helix-loop-helix structure acting as a solid "pushrod" to push the rotor γ subunit, but the short helix in F 1(1/2)s would fail to accomplish this task. (ii) Three different half-torque F 0F 1(1/2)s were purified and reconstituted into proteoliposomes. They carry out ATP-driven proton pumping and build up the same small transmembrane ΔpH, indicating that the final ΔpH is directly related to the amount of torque. (iii) The half-torque F 0F 1(1/2)s can catalyzeATPsynthesis, although slowly. The rate of synthesis varies widely among the three F 0F 1(1/2)s, which suggests that the rate reflects subtle conformational variations of individual mutants.

    Original languageEnglish
    Pages (from-to)1884-1891
    Number of pages8
    JournalJournal of Biological Chemistry
    Volume287
    Issue number3
    DOIs
    Publication statusPublished - 2012 Jan 13

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    Torque
    Adenosine Triphosphate
    Protons
    Enzymes
    Rotors
    Stators
    Catalytic Domain

    ASJC Scopus subject areas

    • Biochemistry
    • Cell Biology
    • Molecular Biology

    Cite this

    Torque generation and utilization in motor enzyme F 0F 1-ATP synthase : Half-torque F 1 with short-sized pushrod helix and reduced ATP synthesis by half-torque F 0F 1. / Usukura, Eiji; Suzuki, Toshiharu; Furuike, Shou; Soga, Naoki; Saita, Ei Ichiro; Hisabori, Toru; Kinosita, Kazuhiko; Yoshida, Masasuke.

    In: Journal of Biological Chemistry, Vol. 287, No. 3, 13.01.2012, p. 1884-1891.

    Research output: Contribution to journalArticle

    Usukura, Eiji ; Suzuki, Toshiharu ; Furuike, Shou ; Soga, Naoki ; Saita, Ei Ichiro ; Hisabori, Toru ; Kinosita, Kazuhiko ; Yoshida, Masasuke. / Torque generation and utilization in motor enzyme F 0F 1-ATP synthase : Half-torque F 1 with short-sized pushrod helix and reduced ATP synthesis by half-torque F 0F 1. In: Journal of Biological Chemistry. 2012 ; Vol. 287, No. 3. pp. 1884-1891.
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    abstract = "ATP synthase (F 0F 1) is made of two motors, a proton-driven motor (F 0) and an ATP-driven motor (F 1), connected by a common rotary shaft, and catalyzes proton flow-driven ATP synthesis and ATP-driven proton pumping. In F 1, the central γ subunit rotates inside the α 3β 3 ring. Here we report structural features of F 1 responsible for torque generation and the catalytic ability of the low-torque F 0F 1. (i) Deletion of one or two turns in the α-helix in the C-terminal domain of catalytic β subunit at the rotor/stator contact region generates mutant F 1s, termed F 1(1/2)s, that rotate with about half of the normal torque. This helix would support the helix-loop-helix structure acting as a solid {"}pushrod{"} to push the rotor γ subunit, but the short helix in F 1(1/2)s would fail to accomplish this task. (ii) Three different half-torque F 0F 1(1/2)s were purified and reconstituted into proteoliposomes. They carry out ATP-driven proton pumping and build up the same small transmembrane ΔpH, indicating that the final ΔpH is directly related to the amount of torque. (iii) The half-torque F 0F 1(1/2)s can catalyzeATPsynthesis, although slowly. The rate of synthesis varies widely among the three F 0F 1(1/2)s, which suggests that the rate reflects subtle conformational variations of individual mutants.",
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    AU - Usukura, Eiji

    AU - Suzuki, Toshiharu

    AU - Furuike, Shou

    AU - Soga, Naoki

    AU - Saita, Ei Ichiro

    AU - Hisabori, Toru

    AU - Kinosita, Kazuhiko

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