Temperature‐sensitive reaction intermediate of F<sub>1</sub>‐ATPase

Watanabe, Rikiya; Iino, Ryota; Shimabukuro, Katsuya; Yoshida, Masasuke; Noji, Hiroyuki

doi:10.1038/sj.embor.7401135

Cited by 47 publications

(72 citation statements)

References 21 publications

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“…Single Molecule Rotation Assays-Single molecule rotation of TA2F 1 ␥2c was measured as described previously using either a ϳ200-nm magnetic bead duplex (high load probe) or a 40-nm gold nanoparticle (low load probe) (33,44,55,78). Briefly, a flow cell was constructed using a Ni 2ϩ -NTA-modified glass surface and an uncoated cover glass (Matsunami Glass Ltd., Japan), and biotinylated TA2F 1 ␥2c subunit complexes were immobilized on the Ni 2ϩ -NTA surface via a hexahistidine tag introduced at the N terminus of the ␤-subunit.…”

Section: Methodsmentioning

confidence: 99%

Biophysical Characterization of a Thermoalkaliphilic Molecular Motor with a High Stepping Torque Gives Insight into Evolutionary ATP Synthase Adaptation

McMillan

Watanabe

Ueno

et al. 2016

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

Biophysical Characterization of a Thermoalkaliphilic Molecular Motor with a High Stepping Torque Gives Insight into Evolutionary ATP Synthase Adaptation

McMillan

Watanabe

Ueno

et al. 2016

Journal of Biological Chemistry

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“…3C) transitions, the transition timings between state I and state II are affected only slightly by the concentration changes of the substrates. It has been shown that the angular position of ATP binding, ADP release, and P i release are around 0°, 0°(or somewhere between 0°and 80°), and 80°, respectively (4,6,24). Taking into account this rotation scheme, it seems that the transition between state III and state I is caused by binding/release of ATP/ADP and the transition between state II to state III is governed by P i release/binding.…”

Section: Magnetic Field Rotationmentioning

confidence: 99%

Simple mechanism whereby the F ₁ -ATPase motor rotates with near-perfect chemomechanical energy conversion

Saita

Suzuki

Kinosita

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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F 1 -ATPase is a motor enzyme in which a central shaft γ subunit rotates 120°per ATP in the cylinder made of α 3 β 3 subunits. During rotation, the chemical energy of ATP hydrolysis (ΔG ATP ) is converted almost entirely into mechanical work by an elusive mechanism. We measured the force for rotation (torque) under various ΔG ATP conditions as a function of rotation angles of the γ subunit with quasistatic, single-molecule manipulation and estimated mechanical work (torque × traveled angle) from the area of the function. The torque functions show three sawtooth-like repeats of a steep jump and linear descent in one catalytic turnover, indicating a simple physical model in which the motor is driven by three springs aligned along a 120°rotation angle. Although the second spring is unaffected by ΔG ATP , activation of the first spring (timing of the torque jump) delays at low [ATP] (or high [ADP]) and activation of the third spring delays at high [P i ]. These shifts decrease the size and area of the sawtooth (magnitude of the work). Thus, F 1 -ATPase responds to the change of ΔG ATP by shifting the torque jump timing and uses ΔG ATP for the mechanical work with near-perfect efficiency. T he F o F 1 -ATP synthase (F o F 1 ) is a ubiquitous enzyme located in bacterial plasma membranes, mitochondrial inner membranes, and chloroplast thylakoid membranes. It plays a critical role in energy metabolism by synthesizing ATP from ADP and inorganic phosphate (P i ). This enzyme consists of and is separable into two major portions: membrane-embedded F o and water-soluble F 1 . In the simplest version of bacterial F o F 1 such as F o F 1 s from thermophilic Bacillus PS3 and Escherichia coli, subunit compositions of F 1 and F o are, respectively, α 3 β 3 γδe and ab 2 c 10 . Both portions are rotary motors that share a common rotor shaft γec 10 . Downward proton flow through F o along the gradient of the electrochemical potential of the proton across the membrane drives the rotation of the c 10 rotor ring in F o that drags rotation of the γe rotor shaft of F 1 in the surrounding α 3 β 3 cylinder. This rotation causes cyclic conformational changes in each of the three catalytic β subunits that result in ATP synthesis (1-3).The isolated F 1 , often called F 1 -ATPase, catalyzes the ATP hydrolysis reaction that drives the rotation of γe to the direction opposite to that in the ATP synthesis. The minimum subunit composition as an ATPase rotary motor is α 3 β 3 γ, and we refer to this complex hereinafter as F 1 . The γ rotates 120°per net hydrolysis of one ATP. Extensive studies, mainly on F 1 from thermophilic Bacillus PS3, have established the nearly complete catalytic scheme of the ATP-driven rotation: Starting from the ATP-waiting state where the orientation angle of the γ is set as 0°, ATP binding to the first β induces the 80°-step rotation of the γ, ADP-release from the second β occurs at some point during this step rotation, and the previously bound ATP in the third β is hydrolyzed at 80°. Then P i release from either the se...

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“…Dwells before the 80°and 40°substeps are termed the binding dwell and catalytic dwell, respectively. ADP release and P i release occur before (or during) the 80°and 40°substeps, respectively (47,48). States in catalytic dwell (D-F) correspond to the crystal structures of F 1 (51).…”

Section: Mechanism Of Inhibition Of Atp Synthesis By the C-terminalmentioning

confidence: 99%

Mechanism of Inhibition by C-terminal α-Helices of the ϵ Subunit of Escherichia coli FoF1-ATP Synthase

Iino

Hasegawa

Tabata

et al. 2009

Journal of Biological Chemistry

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The ⑀ subunit of bacterial F o F 1 -ATP synthase (F o F 1 ), a rotary motor protein, is known to inhibit the ATP hydrolysis reaction of this enzyme. The inhibitory effect is modulated by the conformation of the C-terminal ␣-helices of ⑀, and the "extended" but not "hairpin-folded" state is responsible for inhibition. Although the inhibition of ATP hydrolysis by the C-terminal domain of ⑀ has been extensively studied, the effect on ATP synthesis is not fully understood. In this study, we generated an Escherichia coli

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Temperature‐sensitive reaction intermediate of F₁‐ATPase

Cited by 47 publications

References 21 publications

Biophysical Characterization of a Thermoalkaliphilic Molecular Motor with a High Stepping Torque Gives Insight into Evolutionary ATP Synthase Adaptation

Biophysical Characterization of a Thermoalkaliphilic Molecular Motor with a High Stepping Torque Gives Insight into Evolutionary ATP Synthase Adaptation

Simple mechanism whereby the F ₁ -ATPase motor rotates with near-perfect chemomechanical energy conversion

Mechanism of Inhibition by C-terminal α-Helices of the ϵ Subunit of Escherichia coli FoF1-ATP Synthase

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Temperature‐sensitive reaction intermediate of F1‐ATPase

Cited by 47 publications

References 21 publications

Biophysical Characterization of a Thermoalkaliphilic Molecular Motor with a High Stepping Torque Gives Insight into Evolutionary ATP Synthase Adaptation

Biophysical Characterization of a Thermoalkaliphilic Molecular Motor with a High Stepping Torque Gives Insight into Evolutionary ATP Synthase Adaptation

Simple mechanism whereby the F 1 -ATPase motor rotates with near-perfect chemomechanical energy conversion

Mechanism of Inhibition by C-terminal α-Helices of the ϵ Subunit of Escherichia coli FoF1-ATP Synthase

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Temperature‐sensitive reaction intermediate of F₁‐ATPase

Simple mechanism whereby the F ₁ -ATPase motor rotates with near-perfect chemomechanical energy conversion