Comparison of Brownian-dynamics-based estimates of polymer tension with direct force measurements.

With the aid of brownian dynamics models, it is possible to estimate polymer tension by monitoring polymers' transverse thermal fluctuations. To assess the precision of the approach, brownian dynamics-based tension estimates were compared with the force applied to rhodamine-phalloidin labeled actin filaments bound to polymer beads and suspended between two optical traps. The transverse thermal fluctuations of each filament were monitored with a CCD camera, and the images were analyzed to obtain the filament's transverse displacement variance as a function of position along the filament, the filament's tension, and the camera's exposure time.

Single-molecule stepping and structural dynamics of myosin X.

Myosin X is an unconventional myosin with puzzling motility properties. We studied the motility of dimerized myosin X using the single-molecule fluorescence techniques polTIRF, FIONA and Parallax to measure the rotation angles and three-dimensional position of the molecule during its walk. It was found that Myosin X steps processively in a hand-over-hand manner following a left-handed helical path along both single actin filaments and bundles.

Using electrical and optical tweezers to facilitate studies of molecular motors.

Dielectrophoresis was used to stretch and suspend actin filaments across a trench etched between two electrodes patterned on a glass slide. Optical tweezers were used to bring a motor protein-coated bead into close proximity to a pre-selected, suspended actin filament, facilitating the attachment of the myosin-coated bead to the filament. The clearance beneath the filament allowed the bead to move freely along and around its filamentous track, unhindered by solid surfaces.

Confinement and manipulation of actin filaments by electric fields.

When an AC electric field was applied across a small gap between two metal electrodes elevated above a surface, rhodamine-phalloidin-labeled actin filaments were attracted to the gap and became suspended between the two electrodes. The variance of each filament's horizontal, lateral displacement was measured as a function of electric field intensity and position along the filament. markedly decreased as the electric field intensity increased.