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Mechanically driven ATP synthesis by F1-ATPase


ATP, the main biological energy currency, is synthesized from ADP and inorganic phosphate by ATP synthase in an energy-requiring reaction1,2,3. The F1 portion of ATP synthase, also known as F1-ATPase, functions as a rotary molecular motor: in vitro its γ-subunit rotates4 against the surrounding α3β3 subunits5, hydrolysing ATP in three separate catalytic sites on the β-subunits. It is widely believed that reverse rotation of the γ-subunit, driven by proton flow through the associated Fo portion of ATP synthase, leads to ATP synthesis in biological systems1,2,3,6,7. Here we present direct evidence for the chemical synthesis of ATP driven by mechanical energy. We attached a magnetic bead to the γ-subunit of isolated F1 on a glass surface, and rotated the bead using electrical magnets. Rotation in the appropriate direction resulted in the appearance of ATP in the medium as detected by the luciferase–luciferin reaction. This shows that a vectorial force (torque) working at one particular point on a protein machine can influence a chemical reaction occurring in physically remote catalytic sites, driving the reaction far from equilibrium.

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Figure 1: Experimental set-up.
Figure 2: Rotational synthesis in microdroplets.
Figure 3: Rotational synthesis in a flat chamber.
Figure 4: Dependence of synthesis efficiency on the rate of magnet rotation.

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We thank T. Hayakawa and T. Hiruma of Hamamatsu Photonics KK who allowed H.I. to work on this project for more than 6 years; S. Brenner for the idea of using microdroplets; M. Sugai for initial work; M. Shio, members of the former CREST Team 13 and the current Kinosita and Yoshida laboratories for help and advice; I. Mizuno, K. Suzuki, S. Uchiyama and Y. Mizuguchi for the photon-counting system; C. Gosse and H. Miyajima for the magnetic tweezers; K. Abe and K. Rikukawa for microscopy; S. Murakami for luciferase; and H. Umezawa and M. Fukatsu for laboratory management. This work was supported in part by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and Burroughs Wellcome Fund (R.Y.).

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Correspondence to Hiroyasu Itoh.

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Itoh, H., Takahashi, A., Adachi, K. et al. Mechanically driven ATP synthesis by F1-ATPase. Nature 427, 465–468 (2004).

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