Myosin-V stepping kinetics: a molecular model for processivity.

Myosin-V is a molecular motor that moves processively along its actin track. We have used a feedback-enhanced optical trap to examine the stepping kinetics of this movement. By analyzing the distribution of time periods separating discrete approximately 36-nm mechanical steps, we characterize the number and duration of rate-limiting biochemical transitions preceding each such step. These data show that myosin-V is a tightly coupled motor whose cycle time is limited by ADP release. On the basis of these results, we propose a model for myosin-V processivity.

[1]  Mark J. Schnitzer,et al.  Kinesin hydrolyses one ATP per 8-nm step , 1997, Nature.

[2]  J. Spudich,et al.  Single myosin molecule mechanics: piconewton forces and nanometre steps , 1994, Nature.

[3]  M. Geeves,et al.  Interaction of actin and ADP with the head domain of smooth muscle myosin: implications for strain-dependent ADP release in smooth muscle. , 1998, Biochemistry.

[4]  J A Hammer,et al.  Effect of ADP and Ionic Strength on the Kinetic and Motile Properties of Recombinant Mouse Myosin V* , 2000, The Journal of Biological Chemistry.

[5]  S. Block,et al.  Versatile optical traps with feedback control. , 1998, Methods in enzymology.

[6]  J. Spudich,et al.  Detection of sub-8-nm movements of kinesin by high-resolution optical-trap microscopy. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[7]  E. Krementsova,et al.  Kinetic Characterization of a Monomeric Unconventional Myosin V Construct* , 1999, The Journal of Biological Chemistry.

[8]  Amber L. Wells,et al.  The kinetic mechanism of myosin V. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[9]  J. Howard,et al.  Molecular motors: structural adaptations to cellular functions , 1997, Nature.

[10]  Steven M. Block,et al.  Force and velocity measured for single kinesin molecules , 1994, Cell.

[11]  Kurt Thorn,et al.  Staying on Track: Common Features of DNA Helicases and Microtubule Motors , 1998, Cell.

[12]  D. Hackney,et al.  The kinetic cycles of myosin, kinesin, and dynein. , 1996, Annual review of physiology.

[13]  T. Yanagida,et al.  Mechanics of single kinesin molecules measured by optical trapping nanometry. , 1997, Biophysical journal.

[14]  Samara L. Reck-Peterson,et al.  Class V myosins. , 2000, Biochimica et biophysica acta.

[15]  P. Forscher,et al.  Brain myosin-V is a two-headed unconventional myosin with motor activity , 1993, Cell.

[16]  Toshio Yanagida,et al.  A single myosin head moves along an actin filament with regular steps of 5.3 nanometres , 1999, Nature.

[17]  F. Young Biochemistry , 1955, The Indian Medical Gazette.

[18]  S. Chu,et al.  Quantitative measurements of force and displacement using an optical trap. , 1996, Biophysical journal.

[19]  K. Svoboda,et al.  Fluctuation analysis of motor protein movement and single enzyme kinetics. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[20]  M. Schnitzer,et al.  Statistical kinetics of processive enzymes. , 1995, Cold Spring Harbor symposia on quantitative biology.

[21]  Mark J. Schnitzer,et al.  Single kinesin molecules studied with a molecular force clamp , 1999, Nature.

[22]  L. Goldstein,et al.  Bead movement by single kinesin molecules studied with optical tweezers , 1990, Nature.

[23]  R. Cheney [1] Purification and assay of myosin V , 1998 .

[24]  J. Gelles,et al.  Coupling of kinesin steps to ATP hydrolysis , 1997, Nature.

[25]  Matthias Rief,et al.  Myosin-V is a processive actin-based motor , 1999, Nature.

[26]  Michelle D. Wang,et al.  Force and velocity measured for single molecules of RNA polymerase. , 1998, Science.

[27]  Christoph F. Schmidt,et al.  Direct observation of kinesin stepping by optical trapping interferometry , 1993, Nature.

[28]  T. Pollard,et al.  Kinetic characterization of brush border myosin-I ATPase. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[29]  A. Mehta,et al.  Reflections of a lucid dreamer: optical trap design considerations. , 1998, Methods in cell biology.