title: Force-dependent stepping kinetics of myosin-V.
abstract: Myosin-V is a processive two-headed actin-based motor protein involved in many intracellular transport processes. A key question for understanding myosin-V function and the communication between its two heads is its behavior under load. Since in vivo myosin-V colocalizes with other much stronger motors like kinesins, its behavior under superstall forces is especially relevant. We used optical tweezers with a long-range force feedback to study myosin-V motion under controlled external forward and backward loads over its full run length. We find the mean step size remains constant at approximately 36 nm over a wide range of forces from 5 pN forward to 1.5 pN backward load. We also find two force-dependent transitions in the chemomechanical cycle. The slower ADP-release is rate limiting at low loads and depends only weakly on force. The faster rate depends more strongly on force. The stronger force dependence suggests this rate represents the diffusive search of the leading head for its binding site. In contrast to kinesin motors, myosin-V's run length is essentially independent of force between 5 pN of forward to 1.5 pN of backward load. At superstall forces of 5 pN, we observe continuous backward stepping of myosin-V, indicating that a force-driven reversal of the power stroke is possible.
authors.affiliation: Physics Department E22, Technical University Munich, Garching, Germany.
authors.forename: Anabel E-M
authors.initials: AE
authors.lastname: Clemen
authors.forename: Mojca
authors.initials: M
authors.lastname: Vilfan
authors.forename: Johann
authors.initials: J
authors.lastname: Jaud
authors.forename: Junshan
authors.initials: J
authors.lastname: Zhang
authors.forename: Michael
authors.initials: M
authors.lastname: Bärmann
authors.forename: Matthias
authors.initials: M
authors.lastname: Rief
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D055592:Biophysical Phenomena; D001703:Biophysics; D002645:Chickens; D066298:In Vitro Techniques; D007700:Kinetics; D020409:Molecular Motor Proteins / Q000737:chemistry / Q000378:metabolism; D024701:Myosin Type V / Q000737:chemistry* / Q000378:metabolism; D055095:Optics and Photonics / Q000295:instrumentation
chemical_list: D020409:Molecular Motor Proteins; D024701:Myosin Type V