Activation of pausing F1 motor by external force
- Yoko Hirono-Hara*,†,
- Koji Ishizuka†,‡,
- Kazuhiko Kinosita, Jr.§,
- Masasuke Yoshida‡, and
- Hiroyuki Noji*,¶,∥
- *Institute of Industrial Science and ¶Precursory Research for Embryonic Science and Technology, Japan Science and Technology Corporation, University of Tokyo, Tokyo 153-8505, Japan; ‡Chemical Resources Laboratory, Tokyo Institute of Technology, Yokohama 226-8503, Japan; and §Center for Integrative Bioscience, Okazaki National Research Institutes, Okazaki 444-8585, Japan
-
Edited by Paul D. Boyer, University of California, Los Angeles, CA, and approved January 4, 2005 (received for review September 2, 2004)
Abstract
A rotary motor F1, a catalytic part of ATP synthase, makes a 120° step rotation driven by hydrolysis of one ATP, which consists of 80° and 40° substeps initiated by ATP binding and probably by ADP and/or Pi dissociation, respectively. During active rotations, F1 spontaneously fails in ADP release and pauses after a 80° substep, which is called the ADP-inhibited form. In the present work, we found that, when pushed >+40° with magnetic tweezers, the pausing F1 resumes its active rotation after releasing inhibitory ADP. The rate constant of the mechanical activation exponentially increased with the pushed angle, implying that F1 weakens the affinity of its catalytic site for ADP as the angle goes forward. This finding explains not only its unidirectional nature of rotation, but also its physiological function in ATP synthesis; it would readily bind ADP from solution when rotated backward by an Fo motor in the ATP synthase. Furthermore, the mechanical work for the forced rotation was efficiently converted into work for expelling ADP from the catalytic site, supporting the tight coupling between the rotation and catalytic event.
Footnotes
-
↵ ∥ To whom correspondence should be addressed. E-mail: hnoji{at}iis.u-tokyo.ac.jp.
-
↵ † Y.H.-H. and K.I. contributed equally to this work.
-
Author contributions: H.N. designed research; Y.H.-H., K.I., and H.N. performed research; Y.H.-H., K.I., and H.N. analyzed data; and K.K., M.Y., and H.N. wrote the paper.
-
This paper was submitted directly (Track II) to the PNAS office.
- Copyright © 2005, The National Academy of Sciences
