ORCID Profile
0009-0004-7943-3630
Current Organisation
HMU Health and Medical University
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Publisher: Cold Spring Harbor Laboratory
Date: 12-08-2020
DOI: 10.1101/2020.08.11.245910
Abstract: Cytoskeletal motors transform chemical energy into mechanical work to drive essential cellular functions. Optical trapping experiments have provided crucial insights into the operation of these molecular machines under load. However, the throughput of such force spectroscopy experiments is typically limited to one measurement at a time. Here, we introduce a highly-parallel, microfluidics-based method that allows for rapid collection of force-dependent motility parameters of cytoskeletal motors with two orders of magnitude improvement in throughput compared to currently available methods. We apply tunable hydrodynamic forces to stepping kinesin-1 motors via DNA-tethered beads and utilize a large field of view to simultaneously track the velocities, run lengths and interaction times of hundreds of in idual kinesin-1 molecules under varying resisting and assisting loads. Importantly, the 16-μm long DNA tethers between the motors and the beads significantly reduces the vertical component of the applied force pulling the motors away from the microtubule. Our approach is readily applicable to other molecular systems and constitutes a new methodology for parallelized single-molecule force studies on cytoskeletal motors.
Publisher: Wiley
Date: 23-03-2021
Publisher: Elsevier BV
Date: 05-2015
Location: No location found
No related grants have been discovered for Wim Walter.