Schedule May 27, 2011
On the Mechanical Stabilization of Filopodia
Alexandra Zidovska (Harvard Medical School)

Alexandra Zidovska1,2,3 and Erich Sackmann1
1 Physics Department E22, Technical University Munich, Germany
2 Department of Systems Biology, Harvard Medical School
3 School of Engineering and Applied Sciences/Department of Physics, Harvard University

We studied force-induced elongation of filopodia by coupling magnetic tweezers to the tip through the bacterial coat protein invasin, which couples the force generator to the actin bundles (through myosin X), thus impeding the growth of the actin plus end. Single force pulses (15–30 s) with amplitudes between 20 and 600 pN and staircase-like force scenarios (amplitudes, ~50 pN; step widths, 30 s) were applied. In both cases, the responses consist of a fast viscoelastic deflection followed by a linear flow regime. The deflections are reversible after switching off the forces, suggesting a mechanical memory. The elongation velocity exhibits an exponential distribution (half-width 1/2>~0.02 µms-1) and did not increase systematically with the force amplitudes. We estimate the bending modulus (0.4 x 10-23 J m) and the number of actin filaments (~10) by analyzing filopodium bending fluctuations. Sequestering of intracellular Ca2+ by BAPTA caused a strong reduction in the amplitude of elongation, whereas latrunculin A resulted in loss of the elastic response. We attribute the force-independent velocity to the elongation of actin bundles enabled by the force-induced actin membrane uncoupling and the reversibility by the treadmilling mechanism and an elastic response. [Zidovska A. and Sackmann E., 2011, Biophys. J., 100, 1428-1437]

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