Temperature dependences of torque generation and membrane voltage in the bacterial flagellar motor

Journal article

In their natural habitats bacteria are frequently exposed to sudden changes in temperature that have been shown to affect their swimming. With our believed to be new methods of rapid temperature control for single-molecule microscopy, we measured here the thermal response of the Na(+)-driven chimeric motor expressed in Escherichia coli cells. Motor torque at low load (0.35 μm bead) increased linearly with temperature, twofold between 15°C and 40°C, and torque at high load (1.0 μm bead) was independent of temperature, as reported for the H(+)-driven motor. Single cell membrane voltages were measured by fluorescence imaging and these were almost constant (∼120 mV) over the same temperature range. When the motor was heated above 40°C for 1-2 min the torque at high load dropped reversibly, recovering upon cooling below 40°C. This response was repeatable over as many as 10 heating cycles. Both increases and decreases in torque showed stepwise torque changes with unitary size ∼150 pN nm, close to the torque of a single stator at room temperature (∼180 pN nm), indicating that dynamic stator dissociation occurs at high temperature, with rebinding upon cooling. Our results suggest that the temperature-dependent assembly of stators is a general feature of flagellar motors.

Authors: Inoue, Y; Baker, M; Fukuoka, H; Takahashi, H; Berry, R

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Elsevier
• Journal: Biophysical Journal
• Volume: 105
• Issue: 12
• Pages: 2801-2810
• Publication date: 2013-12-01
• DOI: 10.1016/j.bpj.2013.09.061
• EISSN: 1542-0086
• ISSN: 0006-3495
• Language: English
• Keywords: torque; sodium; recombinant proteins; temperature; membrane potentials; Vibrio alginolyticus; bacterial outer membrane proteins; Escherichia coli; bacterial proteins