THE FORCE GENERATED BY A SINGLE KINESIN MOLECULE AGAINST AN ELASTIC LOAD

To probe the mechanism by which the motor protein kinesin moves along microtubules, we have developed a highly sensitive technique for measu ring the force exerted by a single motor molecule. In this technique, one end of a microtubule is attached to the tip of a flexible glass fi ber of calibrated stiffness. The other end of the microtubule makes co ntact with a surface sparsely coated with kinesin, By imaging the tip of the glass fiber on a photodiode detector, displacement of the micro tubule by kinesin through as little as 1 nm can be detected and forces as small as 1 pN resolved. Using this force-fiber apparatus we have c haracterized the mechanical output of this molecular motor, The speed at which a molecule of kinesin moved along the surface of a microtubul e decreased linearly as the elastic force was increased. The force req uired to stop a single kinesin molecule was 5.4 +/- 1.0 pN (mean +/- S D; n = 16), independent of the stiffness of the fiber, the damping fro m the fluid, and whether the ATP concentration was high or low.