Structural asymmetry and kinetic limping of single rotary F-ATP synthases

Hendrik Sielaff, Seiga Yanagisawa, Wayne Frasch, Wolfgang Junge, Michael Börsch

Research output: Contribution to journalReview article

1 Citation (Scopus)

Abstract

F-ATP synthases use proton flow through the FO domain to synthesize ATP in the F1 domain. In Escherichia coli, the enzyme consists of rotor subunits γεc10 and stator subunits (αβ)3δab2. Subunits c10 or (αβ)3 alone are rotationally symmetric. However, symmetry is broken by the b2 homodimer, which together with subunit δa, forms a single eccentric stalk connecting the membrane embedded FO domain with the soluble F1 domain, and the central rotating and curved stalk composed of subunit γε. Although each of the three catalytic binding sites in (αβ)3 catalyzes the same set of partial reactions in the time average, they might not be fully equivalent at any moment, because the structural symmetry is broken by contact with b2δ in F1 and with b2a in FO. We monitored the enzyme’s rotary progression during ATP hydrolysis by three single-molecule techniques: fluorescence video-microscopy with attached actin filaments, Förster resonance energy transfer between pairs of fluorescence probes, and a polarization assay using gold nanorods. We found that one dwell in the three-stepped rotary progression lasting longer than the other two by a factor of up to 1.6. This effect of the structural asymmetry is small due to the internal elastic coupling.

Original languageEnglish (US)
Article number504
JournalMolecules
DOIs
StatePublished - Jan 30 2019
Externally publishedYes

Fingerprint

adenosine triphosphate
Adenosine Triphosphate
asymmetry
Kinetics
kinetics
Fluorescence
progressions
enzymes
Nanotubes
Video Microscopy
broken symmetry
Energy Transfer
Enzymes
Nanorods
Actin Cytoskeleton
Fluorescence Microscopy
Gold
fluorescence
Energy transfer
Escherichia coli

Keywords

  • Cryo-EM structure
  • Elasticity
  • Escherichia coli
  • FOF1 ATP synthase
  • Single-molecule fluorescence
  • Subunit rotation
  • Symmetry

ASJC Scopus subject areas

  • Analytical Chemistry
  • Chemistry (miscellaneous)
  • Molecular Medicine
  • Pharmaceutical Science
  • Drug Discovery
  • Physical and Theoretical Chemistry
  • Organic Chemistry

Cite this

Structural asymmetry and kinetic limping of single rotary F-ATP synthases. / Sielaff, Hendrik; Yanagisawa, Seiga; Frasch, Wayne; Junge, Wolfgang; Börsch, Michael.

In: Molecules, 30.01.2019.

Research output: Contribution to journalReview article

Sielaff, Hendrik ; Yanagisawa, Seiga ; Frasch, Wayne ; Junge, Wolfgang ; Börsch, Michael. / Structural asymmetry and kinetic limping of single rotary F-ATP synthases. In: Molecules. 2019.
@article{8b31468195aa42779e2a0cd6cbe638c7,
title = "Structural asymmetry and kinetic limping of single rotary F-ATP synthases",
abstract = "F-ATP synthases use proton flow through the FO domain to synthesize ATP in the F1 domain. In Escherichia coli, the enzyme consists of rotor subunits γεc10 and stator subunits (αβ)3δab2. Subunits c10 or (αβ)3 alone are rotationally symmetric. However, symmetry is broken by the b2 homodimer, which together with subunit δa, forms a single eccentric stalk connecting the membrane embedded FO domain with the soluble F1 domain, and the central rotating and curved stalk composed of subunit γε. Although each of the three catalytic binding sites in (αβ)3 catalyzes the same set of partial reactions in the time average, they might not be fully equivalent at any moment, because the structural symmetry is broken by contact with b2δ in F1 and with b2a in FO. We monitored the enzyme’s rotary progression during ATP hydrolysis by three single-molecule techniques: fluorescence video-microscopy with attached actin filaments, F{\"o}rster resonance energy transfer between pairs of fluorescence probes, and a polarization assay using gold nanorods. We found that one dwell in the three-stepped rotary progression lasting longer than the other two by a factor of up to 1.6. This effect of the structural asymmetry is small due to the internal elastic coupling.",
keywords = "Cryo-EM structure, Elasticity, Escherichia coli, FOF1 ATP synthase, Single-molecule fluorescence, Subunit rotation, Symmetry",
author = "Hendrik Sielaff and Seiga Yanagisawa and Wayne Frasch and Wolfgang Junge and Michael B{\"o}rsch",
year = "2019",
month = "1",
day = "30",
doi = "10.3390/molecules24030504",
language = "English (US)",
journal = "Molecules",
issn = "1420-3049",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",

}

TY - JOUR

T1 - Structural asymmetry and kinetic limping of single rotary F-ATP synthases

AU - Sielaff, Hendrik

AU - Yanagisawa, Seiga

AU - Frasch, Wayne

AU - Junge, Wolfgang

AU - Börsch, Michael

PY - 2019/1/30

Y1 - 2019/1/30

N2 - F-ATP synthases use proton flow through the FO domain to synthesize ATP in the F1 domain. In Escherichia coli, the enzyme consists of rotor subunits γεc10 and stator subunits (αβ)3δab2. Subunits c10 or (αβ)3 alone are rotationally symmetric. However, symmetry is broken by the b2 homodimer, which together with subunit δa, forms a single eccentric stalk connecting the membrane embedded FO domain with the soluble F1 domain, and the central rotating and curved stalk composed of subunit γε. Although each of the three catalytic binding sites in (αβ)3 catalyzes the same set of partial reactions in the time average, they might not be fully equivalent at any moment, because the structural symmetry is broken by contact with b2δ in F1 and with b2a in FO. We monitored the enzyme’s rotary progression during ATP hydrolysis by three single-molecule techniques: fluorescence video-microscopy with attached actin filaments, Förster resonance energy transfer between pairs of fluorescence probes, and a polarization assay using gold nanorods. We found that one dwell in the three-stepped rotary progression lasting longer than the other two by a factor of up to 1.6. This effect of the structural asymmetry is small due to the internal elastic coupling.

AB - F-ATP synthases use proton flow through the FO domain to synthesize ATP in the F1 domain. In Escherichia coli, the enzyme consists of rotor subunits γεc10 and stator subunits (αβ)3δab2. Subunits c10 or (αβ)3 alone are rotationally symmetric. However, symmetry is broken by the b2 homodimer, which together with subunit δa, forms a single eccentric stalk connecting the membrane embedded FO domain with the soluble F1 domain, and the central rotating and curved stalk composed of subunit γε. Although each of the three catalytic binding sites in (αβ)3 catalyzes the same set of partial reactions in the time average, they might not be fully equivalent at any moment, because the structural symmetry is broken by contact with b2δ in F1 and with b2a in FO. We monitored the enzyme’s rotary progression during ATP hydrolysis by three single-molecule techniques: fluorescence video-microscopy with attached actin filaments, Förster resonance energy transfer between pairs of fluorescence probes, and a polarization assay using gold nanorods. We found that one dwell in the three-stepped rotary progression lasting longer than the other two by a factor of up to 1.6. This effect of the structural asymmetry is small due to the internal elastic coupling.

KW - Cryo-EM structure

KW - Elasticity

KW - Escherichia coli

KW - FOF1 ATP synthase

KW - Single-molecule fluorescence

KW - Subunit rotation

KW - Symmetry

UR - http://www.scopus.com/inward/record.url?scp=85060704997&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85060704997&partnerID=8YFLogxK

U2 - 10.3390/molecules24030504

DO - 10.3390/molecules24030504

M3 - Review article

JO - Molecules

JF - Molecules

SN - 1420-3049

M1 - 504

ER -