Microtubule curvatures under perpendicular electric forces reveal a low persistence length.

The mechanics of microtubules, cylindrical protein filaments that constitute the cytoskeleton, have been well characterized on long length scales. Here, we investigate the persistence length of short (approximately 0.1 microm) ends of microtubules by measuring the trajectories of kinesin-propelled microtubules under perpendicular electric forces. We relate the measured trajectory curvatures to the biased thermal fluctuations of the leading microtubule end, and upon including all electrohydrodynamic forces, we find that the persistence length of the microtubule ends is only 0.08 +/- 0.02 mm. This is significantly shorter than the well established value of approximately 4-8 mm that is measured for long microtubules. Our data are in good agreement with recent theoretical predictions that microtubules mechanically behave as a loose assembly of independent protofilaments on these short length scales.

Electrophoresis of individual microtubules in microchannels.

We use micrometer-sized fluidic channels to confine and measure electrophoresis of freely suspended individual microtubules. We measure orientation-dependent velocities of microtubules and the electro-osmotic flow mobility in our channels to infer the anisotropic electrophoretic mobility of microtubules under physiological conditions. We discuss the difference between electrophoresis and purely hydrodynamic motion and its implications for interpreting mobility measurements. We show that the mobility anisotropy is a factor of 0.83, clearly different from the well known anisotropy factor of 0.5 in Stokes drag coefficients for cylindrical objects. We also show that the velocity is independent of microtubule length, which would be different for hydrodynamic motion. We demonstrate that the electric force on the counterions has important consequences for the interpretation of electrophoresis experiments and that ignoring this can lead to an underestimation of the effective charge by orders of magnitude. From the electrophoresis measurements, we calculate an effective surface-charge density of -36.7 +/- 0.4 mC/m2 for microtubules. Electrophoretic measurements of subtilisin-digested microtubules, which have the negatively charged C termini on the outer surface removed, show a 24% decrease in mobility and, correspondingly, in surface charge, but no change in anisotropy.