Slip sliding away: load-dependence of velocity generated by skeletal muscle myosin molecules in the laser trap.
Skeletal muscle's ability to shorten and lengthen against a load is a fundamental property, presumably reflecting the inherent load-dependence of the myosin molecular motor. Here we report the velocity of a single actin filament translocated by a mini-ensemble of skeletal myosin approximately 8 heads under constant loads up to 15 pN in a laser trap assay. Actin filament velocity decreased with increasing load hyberbolically, with unloaded velocity and stall force differing by a factor of 2 with [ATP] (30 vs. 100 muM). Analysis of actin filament movement revealed that forward motion was punctuated with rapid backward 60-nm slips, with the slip frequency increasing with resistive load. At stall force, myosin-generated forward movement was balanced by backward slips, whereas at loads greater than stall, myosin could no longer sustain forward motion, resulting in negative velocities as in eccentric contractions of whole muscle. Thus, the force-velocity relationship of muscle reflects both the inherent load-dependence of the actomyosin interaction and the balance between forward and reverse motion observed at the molecular level.
Debold
Edward P
EP
Department of Molecular Physiology & Biophysics, University of Vermont, Burlington, VT 05405, USA.
Patlak
Joseph B
JB
Warshaw
David M
DM
D008841:Actin Cytoskeleton / Q000737:chemistry; D000199:Actins / Q000737:chemistry; D000205:Actomyosin / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D002645:Chickens; D007700:Kinetics; D007834:Lasers; D016014:Linear Models; D020409:Molecular Motor Proteins / Q000737:chemistry; D009068:Movement; D018482:Muscle, Skeletal / Q000378:metabolism / Q000473:pathology*; D009218:Myosins / Q000737:chemistry*; D012984:Software; D013696:Temperature; D013997:Time Factors
D000199:Actins; D020409:Molecular Motor Proteins; D000255:Adenosine Triphosphate; D000205:Actomyosin; D009218:Myosins