Stepwise rotation of the γ-subunit of EF o F 1 -ATP synthase during ATP synthesis: a single-molecule FRET approach

2003 ◽  
Author(s):  
Michael Borsch ◽  
Manuel Diez ◽  
Boris Zimmermann ◽  
Matthias Trost ◽  
Stefan Steigmiller ◽  
...  
Keyword(s):  
Atp Synthase ◽  
Single Molecule ◽  
Atp Synthesis ◽  
Single Molecule Fret ◽  
Γ Subunit

scholarly journals An ATP synthase harboring an atypical γ-subunit is involved in ATP synthesis in tomato fruit chromoplasts

2013 ◽  
Vol 74 (1) ◽  
pp. 74-85 ◽  
Author(s):  
Irini Pateraki ◽  
Marta Renato ◽  
Joaquín Azcón-Bieto ◽  
Albert Boronat
Keyword(s):  
Atp Synthase ◽  
Tomato Fruit ◽  
Atp Synthesis ◽  
Γ Subunit

scholarly journals ATP synthase K+- and H+-fluxes drive ATP synthesis and enable mitochondrial K+-‘uniporter’ function: I. Characterization of ion fluxes

Function ◽  
2021 ◽  
Author(s):  
Magdalena Juhaszova ◽  
Evgeny Kobrinsky ◽  
Dmitry B Zorov ◽  
H Bradley Nuss ◽  
Yael Yaniv ◽  
...  
Keyword(s):  
Atp Synthase ◽  
Single Molecule ◽  
Chemical Potential ◽  
Atp Synthesis ◽  
Energy Utilization ◽  
Photon Detection ◽  
Isolated Mitochondria ◽  
Intact Mitochondrion ◽  
Atp Generation ◽  
K Transport

Abstract ATP synthase (F1Fo) synthesizes daily our body's weight in ATP, whose production-rate can be transiently increased several-fold to meet changes in energy utilization. Using purified mammalian F1Fo-reconstituted proteoliposomes and isolated mitochondria, we show F1Fo can utilize both ΔΨm-driven H+- and K+-transport to synthesize ATP under physiological pH = 7.2 and K+ = 140 mEq/L conditions. Purely K+-driven ATP synthesis from single F1Fo molecules measured by bioluminescence photon detection could be directly demonstrated along with simultaneous measurements of unitary K+ currents by voltage clamp, both blocked by specific Fo inhibitors. In the presence of K+, compared to osmotically-matched conditions in which this cation is absent, isolated mitochondria display 3.5-fold higher rates of ATP synthesis, at the expense of 2.6-fold higher rates of oxygen consumption, these fluxes being driven by a 2.7:1 K+:H+ stoichiometry. The excellent agreement between the functional data obtained from purified F1Fo single molecule experiments and ATP synthase studied in the intact mitochondrion under unaltered OxPhos coupling by K+ presence, is entirely consistent with K+ transport through the ATP synthase driving the observed increase in ATP synthesis. Thus, both K+ (harnessing ΔΨm) and H+ (harnessing its chemical potential energy, ΔµH) drive ATP generation during normal physiology.

3D-localization of the a -subunit in F 0 F 1 -ATP synthase by time resolved single-molecule FRET

2006 ◽  
Author(s):  
Monika G. Düser ◽  
Nawid Zarrabi ◽  
Yumin Bi ◽  
Boris Zimmermann ◽  
Stanley D. Dunn ◽  
...  
Keyword(s):  
Atp Synthase ◽  
Single Molecule ◽  
Time Resolved ◽  
Single Molecule Fret ◽  
3D Localization ◽  
A Subunit

scholarly journals ATP Synthase with Its γ Subunit Reduced to the N-terminal Helix Can Still Catalyze ATP Synthesis

2009 ◽  
Vol 284 (39) ◽  
pp. 26519-26525 ◽  
Author(s):  
Nelli Mnatsakanyan ◽  
Jonathon A. Hook ◽  
Leah Quisenberry ◽  
Joachim Weber
Keyword(s):  
Atp Synthase ◽  
Atp Synthesis ◽  
Γ Subunit

scholarly journals The phototroph-specific β-hairpin structure of the γ subunit of FoF1-ATP synthase is important for efficient ATP-synthesis of cyanobacteria

2021 ◽  
pp. 101027
Author(s):  
Kumiko Kondo ◽  
Masayuki Izumi ◽  
Kosuke Inabe ◽  
Keisuke Yoshida ◽  
Mari Imashimizu ◽  
...  
Keyword(s):  
Atp Synthase ◽  
Atp Synthesis ◽  
Hairpin Structure ◽  
Γ Subunit ◽  
Fof1 Atp Synthase

scholarly journals Stepwise rotation of the γ-subunit of EF0F1-ATP synthase observed by intramolecular single-molecule fluorescence resonance energy transfer1

FEBS Letters ◽  
2002 ◽  
Vol 527 (1-3) ◽  
pp. 147-152 ◽  
Author(s):  
Michael Börsch ◽  
Manuel Diez ◽  
Boris Zimmermann ◽  
Rolf Reuter ◽  
Peter Gräber
Keyword(s):  
Atp Synthase ◽  
Single Molecule ◽  
Resonance Energy ◽  
Single Molecule Fluorescence ◽  
Γ Subunit ◽  
Fluorescence Resonance

C-Terminal Mutations in the Chloroplast ATP Synthase γ Subunit Impair ATP Synthesis and Stimulate ATP Hydrolysis†

Biochemistry ◽  
2008 ◽  
Vol 47 (2) ◽  
pp. 836-844 ◽  
Author(s):  
Feng He ◽  
Hardeep S. Samra ◽  
Eric A. Johnson ◽  
Nicholas R. Degner ◽  
Richard E. McCarty ◽  
...  
Keyword(s):  
Atp Synthase ◽  
Atp Hydrolysis ◽  
Atp Synthesis ◽  
Chloroplast Atp Synthase ◽  
Γ Subunit

scholarly journals Elasticity, friction, and pathway of γ-subunit rotation in FoF1-ATP synthase

2015 ◽  
Vol 112 (34) ◽  
pp. 10720-10725 ◽  
Author(s):  
Kei-ichi Okazaki ◽  
Gerhard Hummer
Keyword(s):  
Atp Synthase ◽  
Single Molecule ◽  
Viscoelastic Model ◽  
Thermodynamic Efficiency ◽  
Γ Subunit ◽  
Frictional Dissipation ◽  
Rotary Motors ◽  
Subunit Rotation ◽  
Dynamics Simulations ◽  
Rotary Motor

We combine molecular simulations and mechanical modeling to explore the mechanism of energy conversion in the coupled rotary motors of FoF1-ATP synthase. A torsional viscoelastic model with frictional dissipation quantitatively reproduces the dynamics and energetics seen in atomistic molecular dynamics simulations of torque-driven γ-subunit rotation in the F1-ATPase rotary motor. The torsional elastic coefficients determined from the simulations agree with results from independent single-molecule experiments probing different segments of the γ-subunit, which resolves a long-lasting controversy. At steady rotational speeds of ∼1 kHz corresponding to experimental turnover, the calculated frictional dissipation of less than kBT per rotation is consistent with the high thermodynamic efficiency of the fully reversible motor. Without load, the maximum rotational speed during transitions between dwells is reached at ∼1 MHz. Energetic constraints dictate a unique pathway for the coupled rotations of the Fo and F1 rotary motors in ATP synthase, and explain the need for the finer stepping of the F1 motor in the mammalian system, as seen in recent experiments. Compensating for incommensurate eightfold and threefold rotational symmetries in Fo and F1, respectively, a significant fraction of the external mechanical work is transiently stored as elastic energy in the γ-subunit. The general framework developed here should be applicable to other molecular machines.

scholarly journals Fast ATP-dependent subunit rotation in reconstituted FoF1-ATP synthase trapped in solution

2021 ◽  
Author(s):  
Thomas Heitkamp ◽  
Michael Börsch
Keyword(s):  
Atp Synthase ◽  
Single Molecule ◽  
Atp Hydrolysis ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Atp Synthesis ◽  
Electrochemical Potential ◽  
Ensemble Averaging ◽  
Subunit Rotation ◽  
Atp Synthases

ABSTRACTFoF1-ATP synthases are the ubiquitous membrane enzymes which catalyze ATP synthesis or ATP hydrolysis in reverse, respectively. Enzyme kinetics are controlled by internal subunit rotation, by substrate and product concentrations, by mechanical inhibitory mechanisms, but also by the electrochemical potential of protons across the membrane. By utilizing an Anti- Brownian electrokinetic trap (ABEL trap), single-molecule Förster resonance energy transfer (smFRET)-based subunit rotation monitoring was prolonged from milliseconds to seconds. The extended observation times for single proteoliposomes in solution allowed to observe fluctuating rotation rates of individual enzymes and to map the broad distributions of ATP-dependent catalytic rates in FoF1-ATP synthase. The buildup of an electrochemical potential of protons was confirmed to limit the maximum rate of ATP hydrolysis. In the presence of ionophores and uncouplers the fastest subunit rotation speeds measured in single reconstituted FoF1-ATP synthases were 180 full rounds per second, i.e. much faster than measured by biochemical ensemble averaging, but not as fast as the maximum rotational speed reported previously for isolated single F1 fragments without coupling to the membrane-embedded Fo domain of the enzyme.

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