scholarly journals Visualizing movements in E. coli F1Fo ATP synthase indicates how the F1 and Fo motors are coupled

2019 ◽  
Author(s):  
Meghna Sobti ◽  
James L. Walshe ◽  
Robert Ishmukhametov ◽  
Alastair G. Stewart
Keyword(s):  
Atp Synthase ◽  
Conformational Changes ◽  
Energy Production ◽  
Structural Model ◽  
Minimal Energy ◽  
F1fo Atp Synthase ◽  
E Coli ◽  
Cellular Energy ◽  
Central Stalk ◽  
Flow Through

AbstractF1Fo ATP synthase functions as a biological rotary generator and makes a major contribution to cellular energy production. It is comprised of two motors that are coupled together by a central “rotor” and peripheral “stator” stalk. Proton flow through the Fo motor generates rotation of the central stalk that induces conformation changes that catalyze production of ATP in the F1 motor. Here we provide 3-4 Å resolution cryo-EM structures of E. coli F1Fo ATP synthase in 10 mM MgADP. In addition to generating a comprehensive structural model of E. coli F1Fo ATP synthase to provide a framework to interpret mutagenesis studies, we describe a rotational sub-step of the Fo motor c-ring associated with long-range conformational changes that suggests an elegant mechanism by which the F1 and Fo motors can be coupled with minimal energy loss.

scholarly journals Subunit epsilon of E. coli F1Fo ATP synthase attenuates enzyme activity by modulating central stalk flexibility

2020 ◽  
Author(s):  
Meghna Sobti ◽  
James L. Walshe ◽  
Yi C. Zeng ◽  
Robert Ishmukhametov ◽  
Alastair G. Stewart
Keyword(s):  
Atp Synthase ◽  
Conformational Changes ◽  
Energy Production ◽  
Potential Role ◽  
F1fo Atp Synthase ◽  
Atp Production ◽  
E Coli ◽  
Cellular Energy ◽  
Central Stalk ◽  
Flow Through

ABSTRACTF1Fo ATP synthase functions as a biological rotary generator that makes a major contribution to cellular energy production. Proton flow through the Fo motor generates rotation of the central stalk, inducing conformational changes in the F1 motor that catalyzes ATP production via flexible coupling. Here we present a range of cryo-EM structures of E. coli ATP synthase in different rotational and inhibited states observed following a 45 second incubation with 10 mM MgATP. The structures generated describe multiple changes that occur following addition of MgATP, with the inhibitory C-terminal domain of subunit ε (εCTD) disassociating from the central stalk to adopt a condensed “down” conformation. The transition to the εCTD down state increases the torsional flexibility of the central stalk allowing its foot to rotate by ∼50°, with further flexing in the peripheral stalk enabling the c-ring to rotate by two sub-steps in the Fo motor. Truncation mutants lacking the second helix of the εCTD suggest that central stalk rotational flexibility is important for F1Fo ATP synthase function. Overall this study identifies the potential role played by torsional flexing within the rotor and how this could be influenced by the ε subunit.

scholarly journals Heterologous expression, purification and crystallization on central stalk and peripheral stalk of F1FO ATP synthase of Aquifex aeolicus

2014 ◽  
Vol 1837 ◽  
pp. e22
Author(s):  
Shuai Xin ◽  
Chunli Zhang ◽  
Julian Langer ◽  
Guohong Peng ◽  
Hartmut Michel
Keyword(s):  
Heterologous Expression ◽  
Atp Synthase ◽  
Aquifex Aeolicus ◽  
F1fo Atp Synthase ◽  
Peripheral Stalk ◽  
Central Stalk

scholarly journals F0-F1 coupling and symmetry mismatch in ATP synthase resolved in every F0 rotation step

2021 ◽  
Author(s):  
Shintaroh Kubo ◽  
Toru Niina ◽  
Shoji Takada
Keyword(s):  
Electron Microscopy ◽  
Atp Synthase ◽  
Energy Production ◽  
Comprehensive Understanding ◽  
Cellular Energy ◽  
Cryo Electron Microscopy ◽  
Rotary Motors ◽  
Elastic Distortion ◽  
Dynamics Simulations ◽  
Symmetric Structures

The F0F1 ATP synthase, essential for cellular energy production, is composed of the F0 and F1 rotary motors. While both F0 and F1 have pseudo-symmetric structures, their symmetries do not match. How the symmetry mismatch is solved remains elusive due to missing intermediate structures of rotational steps. Here, for ATP synthases with 3- and 10-fold symmetries in F1 and F0, respectively, we uncovered the mechanical couplings between F0 and F1 at every 36° rotation step via molecular dynamics simulations and comparison of cryo-electron microscopy structures from three species. We found that the frustration is shared by several elements. The F1 stator partially rotates relative to the F0 stator via elastic distortion of the b-subunits. The rotor can be distorted. The c-ring rotary angles can be deviated from symmetric ones. Additionally, the F1 motor may take non-canonical structures relieving stronger frustration. Together, we provide comprehensive understanding to solve the symmetry mismatch.

scholarly journals Expression, purification and crystallization of central stalk and peripheral stalk of F1Fo ATP synthase from Aquifex aeolicus in Escherichia coli

2012 ◽  
Vol 1817 ◽  
pp. S27
Author(s):  
Shuai Xin ◽  
Chunli Zhang ◽  
Julian Langer ◽  
Guohong Peng ◽  
Hartmut Michel
Keyword(s):  
Escherichia Coli ◽  
Atp Synthase ◽  
Aquifex Aeolicus ◽  
F1fo Atp Synthase ◽  
Peripheral Stalk ◽  
Central Stalk

scholarly journals E. Coli F1Fo ATP Synthase Subunit C: Solution NMR Structure by CS-Rosetta

2013 ◽  
Vol 104 (2) ◽  
pp. 179a
Author(s):  
Yisong Tao ◽  
Mark Girvin
Keyword(s):  
Atp Synthase ◽  
Nmr Structure ◽  
Solution Nmr ◽  
Subunit C ◽  
F1fo Atp Synthase ◽  
E Coli

scholarly journals Fluidity of Structure and Swiveling of Helices in the Subunit c Ring of Escherichia coli ATP Synthase as Revealed by Cysteine-Cysteine Cross-Linking

2007 ◽  
Vol 282 (46) ◽  
pp. 33788-33794 ◽  
Author(s):  
Owen D. Vincent ◽  
Brian E. Schwem ◽  
P. Ryan Steed ◽  
Warren Jiang ◽  
Robert H. Fillingame
Keyword(s):  
Escherichia Coli ◽  
Atp Synthase ◽  
Time Course ◽  
Structural Model ◽  
Ring Structure ◽  
High Yield ◽  
Cross Linking ◽  
E Coli ◽  
The Cross ◽  
Ilyobacter Tartaricus

Subunit c in the membrane-traversing F0 sector of Escherichia coli ATP synthase is known to fold with two transmembrane helices and form an oligomeric ring of 10 or more subunits in the membrane. Models for the E. coli ring structure have been proposed based upon NMR solution structures and intersubunit cross-linking of Cys residues in the membrane. The E. coli models differ from the recent x-ray diffraction structure of the isolated Ilyobacter tartaricus c-ring. Furthermore, key cross-linking results supporting the E. coli model prove to be incompatible with the I. tartaricus structure. To test the applicability of the I. tartaricus model to the E. coli c-ring, we compared the cross-linking of a pair of doubly Cys substituted c-subunits, each of which was compatible with one model but not the other. The key finding of this study is that both A21C/M65C and A21C/I66C doubly substituted c-subunits form high yield oligomeric structures, c2, c3... c10, via intersubunit disulfide bond formation. The results indicate that helical swiveling, with resultant interconversion of the two conformers predicted by the E. coli and I. tartaricus models, must be occurring over the time course of the cross-linking experiment. In the additional experiments reported here, we tried to ascertain the preferred conformation in the membrane to help define the most likely structural model. We conclude that both structures must be able to form in the membrane, but that the helical swiveling that promotes their interconversion may not be necessary during rotary function.

scholarly journals Cytoplasmic Loops of Subunits C and A in E. Coli F1Fo ATP Synthase Interact to Gate H+ Transport to the Cytoplasm

2014 ◽  
Vol 106 (2) ◽  
pp. 372a
Author(s):  
Robert H. Fillingame
Keyword(s):  
Atp Synthase ◽  
F1fo Atp Synthase ◽  
E Coli ◽  
Cytoplasmic Loops

scholarly journals Cryo-EM structures provide insight into how E. coli F1Fo ATP synthase accommodates symmetry mismatch

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Meghna Sobti ◽  
James L. Walshe ◽  
Di Wu ◽  
Robert Ishmukhametov ◽  
Yi C. Zeng ◽  
...  
Keyword(s):  
Atp Synthase ◽  
F1fo Atp Synthase ◽  
E Coli ◽  
Insight Into

scholarly journals Heterologous expression of the assembled and functional F1FO ATP synthase from A. aeolicus in E. Coli

2012 ◽  
Vol 1817 ◽  
pp. S28
Author(s):  
Chunli Zhang ◽  
Julian Langer ◽  
Matteo Allegretti ◽  
Janet Vonck ◽  
Guohong Peng ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Atp Synthase ◽  
F1fo Atp Synthase ◽  
E Coli

scholarly journals Spotlighting motors and controls of single FoF1-ATP synthase

2013 ◽  
Vol 41 (5) ◽  
pp. 1219-1226 ◽  
Author(s):  
Michael Börsch ◽  
Thomas M. Duncan
Keyword(s):  
Escherichia Coli ◽  
Atp Synthase ◽  
Single Molecule ◽  
Conformational Changes ◽  
Structural Information ◽  
E Coli ◽  
Single Molecule Fret ◽  
Membrane Enzyme ◽  
Subunit Rotation ◽  
Fof1 Atp Synthase

Subunit rotation is the mechanochemical intermediate for the catalytic activity of the membrane enzyme FoF1-ATP synthase. smFRET (single-molecule FRET) studies have provided insights into the step sizes of the F1 and Fo motors, internal transient elastic energy storage and controls of the motors. To develop and interpret smFRET experiments, atomic structural information is required. The recent F1 structure of the Escherichia coli enzyme with the ϵ-subunit in an inhibitory conformation initiated a study for real-time monitoring of the conformational changes of ϵ. The present mini-review summarizes smFRET rotation experiments and previews new smFRET data on the conformational changes of the CTD (C-terminal domain) of ϵ in the E. coli enzyme.

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