Quantification of flagellar motor stator dynamics through in vivo proton-motive force control.

The bacterial flagellar motor, one of the few rotary motors in nature, produces torque to drive the flagellar filament by ion translocation through membrane-bound stator complexes. We used the light-driven proton pump proteorhodopsin (pR) to control the proton-motive force (PMF) in vivo by illuminat...

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Main Authors: Tipping, M, Steel, B, Delalez, N, Berry, R, Armitage, J
Format: Journal article
Language:English
Published: 2013
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author Tipping, M
Steel, B
Delalez, N
Berry, R
Armitage, J
author_facet Tipping, M
Steel, B
Delalez, N
Berry, R
Armitage, J
author_sort Tipping, M
collection OXFORD
description The bacterial flagellar motor, one of the few rotary motors in nature, produces torque to drive the flagellar filament by ion translocation through membrane-bound stator complexes. We used the light-driven proton pump proteorhodopsin (pR) to control the proton-motive force (PMF) in vivo by illumination. pR excitation was shown to be sufficient to replace native PMF generation, and when excited in cells with intact native PMF generation systems increased motor speed beyond the physiological norm. We characterized the effects of rapid in vivo PMF changes on the flagellar motor. Transient PMF disruption events from loss of illumination caused motors to stop, with rapid recovery of their previous rotation rate after return of illumination. However, extended periods of PMF loss led to stepwise increases in rotation rate upon PMF return as stators returned to the motor. The rate constant for stator binding to a putative single binding site on the motor was calculated to be 0.06 s(-1). Using GFP-tagged MotB stator proteins, we found that transient PMF disruption leads to reversible stator diffusion away from the flagellar motor, showing that PMF presence is necessary for continued motor integrity, and calculated a stator dissociation rate of 0.038 s(-1).
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spelling oxford-uuid:8e8b7984-d71a-4853-bff6-ce0ac7a419832022-03-26T22:58:30ZQuantification of flagellar motor stator dynamics through in vivo proton-motive force control.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:8e8b7984-d71a-4853-bff6-ce0ac7a41983EnglishSymplectic Elements at Oxford2013Tipping, MSteel, BDelalez, NBerry, RArmitage, JThe bacterial flagellar motor, one of the few rotary motors in nature, produces torque to drive the flagellar filament by ion translocation through membrane-bound stator complexes. We used the light-driven proton pump proteorhodopsin (pR) to control the proton-motive force (PMF) in vivo by illumination. pR excitation was shown to be sufficient to replace native PMF generation, and when excited in cells with intact native PMF generation systems increased motor speed beyond the physiological norm. We characterized the effects of rapid in vivo PMF changes on the flagellar motor. Transient PMF disruption events from loss of illumination caused motors to stop, with rapid recovery of their previous rotation rate after return of illumination. However, extended periods of PMF loss led to stepwise increases in rotation rate upon PMF return as stators returned to the motor. The rate constant for stator binding to a putative single binding site on the motor was calculated to be 0.06 s(-1). Using GFP-tagged MotB stator proteins, we found that transient PMF disruption leads to reversible stator diffusion away from the flagellar motor, showing that PMF presence is necessary for continued motor integrity, and calculated a stator dissociation rate of 0.038 s(-1).
spellingShingle Tipping, M
Steel, B
Delalez, N
Berry, R
Armitage, J
Quantification of flagellar motor stator dynamics through in vivo proton-motive force control.
title Quantification of flagellar motor stator dynamics through in vivo proton-motive force control.
title_full Quantification of flagellar motor stator dynamics through in vivo proton-motive force control.
title_fullStr Quantification of flagellar motor stator dynamics through in vivo proton-motive force control.
title_full_unstemmed Quantification of flagellar motor stator dynamics through in vivo proton-motive force control.
title_short Quantification of flagellar motor stator dynamics through in vivo proton-motive force control.
title_sort quantification of flagellar motor stator dynamics through in vivo proton motive force control
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AT berryr quantificationofflagellarmotorstatordynamicsthroughinvivoprotonmotiveforcecontrol
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