Smooth muscle and skeletal muscle myosins produce similar unitary forces and displacements in the laser trap.
Purified smooth muscle myosin in the in vitro motility assay propels actin filaments at 1/10 the velocity, yet produces 3-4 times more force than skeletal muscle myosin. At the level of a single myosin molecule, these differences in force and actin filament velocity may be reflected in the size and duration of single motion and force-generating events, or in the kinetics of the cross-bridge cycle. Specifically, an increase in either unitary force or duty cycle may explain the enhanced force-generating capacity of smooth muscle myosin. Similarly, an increase in attached time or decrease in unitary displacement may explain the reduced actin filament velocity of smooth muscle myosin. To discriminate between these possibilities, we used a laser trap to measure unitary forces and displacements from single smooth and skeletal muscle myosin molecules. We analyzed our data using mean-variance analysis, which does not rely on scoring individual events by eye, and emphasizes periods in the data with constant properties. Both myosins demonstrated multiple but similar event populations with discrete peaks at approximately +11 and -11 nm in displacement, and 1.5 and 3.5 pN in force. Mean attached times for smooth muscle myosin were longer than for skeletal-muscle myosin. These results explain much of the difference in actin filament velocity between these myosins, and suggest that an increased duty cycle is responsible for the enhanced force-generating capacity of smooth over skeletal-muscle myosin.
Guilford
W H
WH
Department of Molecular Physiology and Biophysics, University of Vermont, Burlington 05405, USA.
Dupuis
D E
DE
Kennedy
G
G
Wu
J
J
Patlak
J B
JB
Warshaw
D M
DM
D000199:Actins / Q000502:physiology*; D000818:Animals; D001696:Biomechanical Phenomena; D002645:Chickens; D005033:Ethylmaleimide; D005895:Gizzard, Avian; D007537:Isometric Contraction; D007700:Kinetics; D007834:Lasers; D009068:Movement; D018482:Muscle, Skeletal / Q000502:physiology*; D009130:Muscle, Smooth / Q000502:physiology*; D009218:Myosins / Q000737:chemistry* / Q000502:physiology*; D013997:Time Factors; D014422:Turkeys
D000199:Actins; D009218:Myosins; D005033:Ethylmaleimide