Effects of stalk orientation and size of trapped bead on force-velocity relation of kinesin motor determined using single molecule optical trapping methods

Abstract

Conventional kinesin protein is a prototypical biological molecular motor that can step processively on microtubules towards the plus end by hydrolyzing ATP molecules, performing the biological function of intracellular transports. An important characteristic of the kinesin is the load dependence of its velocity, which is usually measured by using the single molecule optical trapping method with a large-sized bead attached to the motor stalk. Puzzlingly, even for the same kinesin, some experiments showed that the velocity is nearly independent of the forward load whereas others showed that the velocity decreases evidently with the increase in the magnitude of the forward load. Here, a theoretical explanation is provided of why different experiments give different dependencies of the velocity on the forward load. It is shown that both the stalk orientation and bead size play a critical role in the different dependencies. Additionally, the reason why the optical trapping experiments with the movable trap usually gave a sigmoid form of the velocity versus backward load whereas with the fixed trap gave a nearly linear form is also explained theoretically. The study is not only critical to the understanding of the response of the motor to the load but also provides strong insights into the coupling mechanism of the motor.

Keywords: Force–velocity form; Kinesin; Molecular motor; Optical trapping assay.