motormax forcemax speedmax power rotary rotary flagellar motor (8 units, e. coli) 2400 pn nm 95 pn...
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Motor Max Force Max Speed Max Power
ROTARY
Flagellar motor (8 units, E. coli)
2400 pN nm
95 pN
300 Hz
35 m/s
2000 pN nm @ 150 Hz
1.9 x 106 pN nm /s
(Vibrio) 1700 Hz
220 m/s
(single unit) 300 pN nm
12 pN
300 Hz
35 m/s
250 pN nm @ 150 Hz
2.4 x 105 pN nm /s
F1 -ATPase 40 pN nm
40 pN
150 Hz
0.9 m/s
20 pN nm @ 75 Hz
9 x 103 pN nm /s
LINEAR
Myosin (single molecule in muscle or in vitro)
6 pN 10 m/s(speed of array, each
molecule mostly detached)
2 pN @ 10 /s x 20 nm
400 pN nm /s
Kinesin 5 pN 1 m/s 2.5 pN @ 0.5 m /s
1.25 x 103 pN nm /s
RNA polymerase
20 pN 0.01 m/s ~200 pN nm /s
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Overview of lectures
• BIOLOGY was introduced by Judy Armitage’s
• BIOPHYSICS - Experimental Techniques to measure rotary molecular motors
Flagellar Motor
F1-ATPase
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Single-molecule experiments on bacterial flagellar motors
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Rotor
Stator
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Continuous switch model
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Switching
F Bai, RW. Branch, D Nicolau, TPilizota, BC Steel, PK Maini, RM Berry (2010)Conformational spread as a mechanism for cooperativity in the bacterial flagellar switchScience 327:685-689
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Bacterial Chemotaxis
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1-D Ising model of flagellar switch
movie
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Tethered cells
Cell body rotates at ~ 10 Hz
1 m
Flagellum tethered to coverslip
Cell body
Coverslip in microscope
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Frequency (Hz)
fast beads, small.mov
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Work = torque x angle
Torque = d(work) / d(angle)
Low Reynolds number:Torque = viscous drag coefficient x angular velocity
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Beads attached to the motor
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Finite, variable switch times
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Switch times distribution predicted by model
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… further detailed tests of model
C / Co
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Resurrection
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resurrection
steady-state expression
One motor can contain at least 11 stators
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1 m bead
0.3 m bead
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Torque-versus speed
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Stepping rotation
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Speed control for step detection using sodium-driven chimaera
pmf or smf = Vm + kT/e ln (Cin/Cout)
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Low numbers of stators:
Low-level induction of stator proteins
De-energization also affects stator number
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Slow rotation with (probably) one stator unit
Back-focal plane detection Fluorescence detection
Real speed 30x slower
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26 steps per revolution
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Kinesin, myosin II, Myosin V, F1-ATPase:
Step size is set by the track
by energy conservation based on full energization and high loads, one proton gives a max step size of ~10 degrees. Maybe there are 2 ions per step?
34-fold model refines C-ring : 25-fold model refines M ring (& C-inner)Thomas et al 2006
One ATP per step
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ATP
ADP+ Pi
10nm
F1
FO
H+ or Na+
Single-molecule experiments on ATP-synthase
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F1
Biotin-avidin link to rotating handle(actin filament, beads)
His-tag link to surface
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0.5 micron beadsRotated by F1
Fluorescent actin filamentRotated by F1
(movie: Wolfgang Junge)
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High [ATP] : no wait before 90° step
Medium [ATP] : ~ms before 90° step
Low [ATP] : long wait before 90° step
(ms)
All [ATP] : ~ms before 30° step
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Low [ATP] : - 90° step rate-limiting- exponential distribution- single step
High [ATP] : - 30° step rate-limiting- peaked distribution- double (or more) step
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