Measuring the number and spacing of molecular motors propelling a gliding microtubule

Abstract

The molecular motor gliding assay, in which a microtubule or other filament moves across a surface coated with motors, has provided much insight into how molecular motors work. The kinesin-microtubule system is also a strong candidate for the job of nanoparticle transporter in nanotechnology devices. In most cases, several motors transport each filament. Each motor serves both to bind the microtubule to a stationary surface and to propel the microtubule along the surface. By applying a uniform transverse force of 4-19 pN to a superparamagnetic bead attached to the trailing end of the microtubule, we have measured the distance d between binding points (motors). The average value of d was determined as a function of motor surface density σ. The measurements agree well with the scaling model of Duke, Holy, and Liebler, which predicts that (d)~σ(-2/5) if 0.05≤σ≤20 μm(-2) [Phys. Rev. Lett. 74, 330 (1995)]. The distribution of d fits an extension of the model. The radius of curvature of a microtubule bent at a binding point by the force of the magnetic bead was ≈1 μm, 5000-fold smaller than the radius of curvature of microtubules subjected only to thermal forces. This is evidence that at these points of high bending stress, generated by the force on the magnetic bead, the microtubule is in the more flexible state of a two-state model of microtubule bending proposed by Heussinger, Schüller, and Frey [Phys. Rev. E 81, 021904 (2010)].