Force generation in single conventional actomyosin complexes under high dynamic load.
The mechanical load borne by a molecular motor affects its force, sliding distance, and its rate of energy transduction. The control of ATPase activity by the mechanical load on a muscle tunes its efficiency to the immediate task, increasing ATP hydrolysis as the power output increases at forces less than isometric (the Fenn effect) and suppressing ATP hydrolysis when the force is greater than isometric. In this work, we used a novel 'isometric' optical clamp to study the mechanics of myosin II molecules to detect the reaction steps that depend on the dynamic properties of the load. An actin filament suspended between two beads and held in separate optical traps is brought close to a surface that is sparsely coated with motor proteins on pedestals of silica beads. A feedback system increases the effective stiffness of the actin by clamping the force on one of the beads and moving the other bead electrooptically. Forces measured during actomyosin interactions are increased at higher effective stiffness. The results indicate that single myosin molecules transduce energy nearly as efficiently as whole muscle and that the mechanical control of the ATP hydrolysis rate is in part exerted by reversal of the force-generating actomyosin transition under high load without net utilization of ATP.
Takagi
Yasuharu
Y
Pennsylvania Muscle Institute, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6392, USA.
Homsher
Earl E
EE
Goldman
Yale E
YE
Shuman
Henry
H
D008841:Actin Cytoskeleton / Q000502:physiology*; D000205:Actomyosin / Q000502:physiology*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001696:Biomechanical Phenomena; D001710:Biotin / Q000737:chemistry; D008863:Microspheres; D020409:Molecular Motor Proteins / Q000502:physiology*; D009010:Monte Carlo Method; D009119:Muscle Contraction / Q000502:physiology*; D024462:Myosin Type II / Q000502:physiology*; D011817:Rabbits
D020409:Molecular Motor Proteins; D001710:Biotin; D000255:Adenosine Triphosphate; D000205:Actomyosin; D024462:Myosin Type II