Direct observation of the rotation of F1-ATPase

  title={Direct observation of the rotation of F1-ATPase},
  author={Hiroyuki Noji and Ryohei Yasuda and Masasuke Yoshida and Kazuhiko Kinosita},
Cells employ a variety of linear motors, such as myosin1–3, kinesin4 and RNA polymerase5, which move along and exert force on a filamentous structure. But only one rotary motor has been investigated in detail, the bacterial flagellum6 (a complex of about 100 protein molecules7). We now show that a single molecule of F1-ATPase acts as a rotary motor, the smallest known, by direct observation of its motion. A central rotor of radius ∼1 nm, formed by its γ-subunit, turns in a stator barrel of… 

Rotary F1-ATPase

The rotation of γ within hours is compatible with the spectroscopically detected blockade of rotation in the AMP-PNP-inhibited enzyme in the time-range of seconds.

Phosphate release coupled to rotary motion of F1-ATPase

Atomistic molecular dynamics simulations are used to construct a first atomistic conformation of the intermediate state following the 40° substep of rotary motion, and to study the timing and molecular mechanism of inorganic phosphate (Pi) release coupled to the rotation.

Single-molecule observation of rotation of F1-ATPase through microbeads.

The relationships between chemical and mechanical events are shown by imaging individual F(1) molecules under an optical microscope and a new scheme emerges: as soon as a catalytic site binds ATP, the gamma-subunit always turns the same face (interaction surface) to the beta hosting that site.

Linear and rotary molecular motors.

  • K. Kinosita
  • Biology, Physics
    Advances in experimental medicine and biology
  • 1998
The use of huge and small probes as described above should be useful in studies of protein machines in general, as a means of detecting conformational changes in a single protein molecule during function.

Single-molecule imaging of rotation of F1-ATPase.

None of the Rotor Residues of F1-ATPase Are Essential for Torque Generation

How subunit coupling produces the γ-subunit rotary motion in F1-ATPase

  • J. PuM. Karplus
  • Biology
    Proceedings of the National Academy of Sciences
  • 2008
A coarse-grained plastic network model is used to show at a residue level of detail how the conformational changes of the catalytic β-subunits act on the γ-subunit through repulsive van der Waals interactions to generate a torque that drives unidirectional rotation, as observed experimentally.

Resolution of distinct rotational substeps by submillisecond kinetic analysis of F1-ATPase

It is shown by high-speed imaging that the 120° step consists of roughly 90° and 30° substeps, each taking only a fraction of a millisecond, which supports the binding-change model for ATP synthesis by reverse rotation of F1-ATPase.

Chemomechanical coupling of human mitochondrial F1-ATPase motor.

ATP-driven rotation of human mitochondrial F1 is reported, demonstrating that chemomechanical coupling angles of the γ-subunit are tuned during evolution.



Intersubunit rotation in active F-ATPase

An intersubunit rotation in real time in the functional enzyme F-ATPase is recorded by applying polarized absorption relaxation after photobleaching to immobilized F1 with eosin-labelled γ in a timespan of 100 ms, compatible with the rate of ATP hydrolysis by immobilization F1.

Rotation of subunits during catalysis by Escherichia coli F1-ATPase.

The results demonstrate that gamma subunit rotates relative to the beta subunits during catalysis, and similar reactivities of unlabeled and radiolabeled beta sub units with gamma C87 upon reoxidation.

Single-molecule analysis of the actomyosin motor using nano-manipulation.

The results suggested that an ATPase cycle produces one power stroke at high load and many ones at low load, similar to those deduced from noise analysis of force fluctuations caused by multiple molecules.

Molecular switch of F0F1-ATP synthase, G-protein, and other ATP-driven enzymes

Probably, binding of nucleotide to F0F1-ATP synthase induces conformational change of the switch II-like region with transforming β subunit structure from “open” to “closed” form and this transformation results in loss of hydrogen bonds with the γ subunit, thus enabling the δ subunit to move.

Nucleotide-dependent Movement of the ε Subunit between α and β Subunits in the Escherichia coli F1F0-type ATPase*

Mutants of ECF1-ATPase were generated, containing cysteine residues in one or more of the following positions: αSer-411, βGlu-381, and εSer-108, after which disulfide bridges could be created by