Nonequivalence of membrane voltage and ion-gradient as driving forces for the bacterial flagellar motor at low load.

Many bacterial species swim using flagella. The flagellar motor couples ion flow across the cytoplasmic membrane to rotation. Ion flow is driven by both a membrane potential (V(m)) and a transmembrane concentration gradient. To investigate their relation to bacterial flagellar motor function we developed a fluorescence technique to measure V(m) in single cells, using the dye tetramethyl rhodamine methyl ester. We used a convolution model to determine the relationship between fluorescence intensity in images of cells and intracellular dye concentration, and calculated V(m) using the ratio of intracellular/extracellular dye concentration. We found V(m) = -140 +/- 14 mV in Escherichia coli at external pH 7.0 (pH(ex)), decreasing to -85 +/- 10 mV at pH(ex) 5.0. We also estimated the sodium-motive force (SMF) by combining single-cell measurements of V(m) and intracellular sodium concentration. We were able to vary the SMF between -187 +/- 15 mV and -53 +/- 15 mV by varying pH(ex) in the range 7.0-5.0 and extracellular sodium concentration in the range 1-85 mM. Rotation rates for 0.35-microm- and 1-microm-diameter beads attached to Na(+)-driven chimeric flagellar motors varied linearly with V(m). For the larger beads, the two components of the SMF were equivalent, whereas for smaller beads at a given SMF, the speed increased with sodium gradient and external sodium concentration.

• Publication year

  2007

• Venue

  Biophysical Journal

• Publication date

  2007-07-01

• Fields of study

  Biology, Medicine, Physics, Chemistry

• Identifiers
  DOI 10.1529/BIOPHYSJ.106.095265PMID 17416615PMCID PMC1914430
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• A programmable optical angle clamp for rotary molecular motors.

  2007cited by this paper
• Fluorescence measurement of intracellular sodium concentration in single Escherichia coli cells.

  2006influential reference
• Torque–speed relationship of the bacterial flagellar motor

  2006cited by this paper
• The maximum number of torque-generating units in the flagellar motor of Escherichia coli is at least 11.

  2006influential reference
• Stoichiometry and turnover in single, functioning membrane protein complexes

  2006cited by this paper
• Direct observation of steps in rotation of the bacterial flagellar motor

  2005influential reference
• Effect of Intracellular pH on Rotational Speed of Bacterial Flagellar Motors

  2003cited by this paper
• Probing the Transmembrane Potential of Bacterial Cells by Voltage-Sensitive Dyes

  2003cited by this paper
• Ion-coupling determinants of Na+-driven and H+-driven flagellar motors.

  2003cited by this paper
• The speed of the flagellar rotary motor of Escherichia coli varies linearly with protonmotive force

  2003influential reference
• The rotary motor of bacterial flagella.

  2003cited by this paper
• Torque-speed relationship of the Na+-driven flagellar motor of Vibrio alginolyticus.

  2003cited by this paper
• Na(+)/H(+) antiporters.

  2001cited by this paper
• Solvent-isotope and pH effects on flagellar rotation in Escherichia coli.

  2000cited by this paper
• Measurement of mitochondrial membrane potential using fluorescent rhodamine derivatives.

  1999cited by this paper
• Accurate flow cytometric membrane potential measurement in bacteria using diethyloxacarbocyanine and a ratiometric technique.

  1999cited by this paper
• The bacterial flagella motor.

  1999cited by this paper
• Transient Mitochondrial Depolarizations Reflect Focal Sarcoplasmic Reticular Calcium Release in Single Rat Cardiomyocytes

  1998cited by this paper
• Intracellular fluorescent probe concentrations by confocal microscopy.

  1998cited by this paper
• Powering the flagellar motor of Escherichia coli with an external voltage source

  1995influential reference
• Imaging in five dimensions: time-dependent membrane potentials in individual mitochondria.

  1993cited by this paper
• Membrane potential can be determined in individual cells from the nernstian distribution of cationic dyes.

  1988cited by this paper
• The stall torque of the bacterial flagellar motor.

  1987cited by this paper
• Coupling between the sodium and proton gradients in respiring Escherichia coli cells measured by 23Na and 31P nuclear magnetic resonance.

  1986cited by this paper
• Constraints on flagellar rotation.

  1985cited by this paper
• Quantitative measurements of membrane potential in Escherichia coli.

  1980cited by this paper
• Energetics of flagellar rotation in bacteria.

  1980cited by this paper
• The Biophysical Journal

  1960cited by this paper
• Bacterial Flagella

  1951cited by this paper

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