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Loading direction regulates the affinity of ADP for kinesin

Nature Structural & Molecular Biology volume 10, pages 308–311 (2003)Cite this article

Abstract

Kinesin is an ATP-driven molecular motor that moves processively along a microtubule. Processivity has been explained as a mechanism that involves alternating single- and double-headed binding of kinesin to microtubules coupled to the ATPase cycle of the motor. The internal load imposed between the two bound heads has been proposed to be a key factor regulating the ATPase cycle in each head. Here we show that external load imposed along the direction of motility on a single kinesin molecule enhances the binding affinity of ADP for kinesin, whereas an external load imposed against the direction of motility decreases it. This coupling between loading direction and enzymatic activity is in accord with the idea that the internal load plays a key role in the unidirectional and cooperative movement of processive motors.

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Figure 1: Measurement of unbinding force.
Figure 2: Unbinding force distributions at various ADP concentrations.
Figure 3: Proportion of weak binding component, W, at various ADP concentrations.
Figure 4: Schematic illustrations showing equilibrium of ADP (D) binding to a complex of kinesin (nucleotide-binding pocket is shown) and microtubule (thin line, where both the plus and the minus ends are shown).

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Acknowledgements

We thank S.S. Rosenfeld and M. Chee for critical reading of the manuscript. We also thank D.D. Hackney and K. Kinosita Jr. for valuable discussion. This research was partly supported by Grants-in-Aid for Specially Promoted Research and for the Bio-venture Project from the Ministry of Education, Sports, Culture, Science and Technology of Japan. S.U. is a recipient of predoctoral fellowship of the Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Department of Physics, School of Science and Engineering, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo, 169-8555, Japan

    Sotaro Uemura & Shin'ichi Ishiwata

  2. Advanced Research Institute for Science and Engineering, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo, 169-8555, Japan

    Shin'ichi Ishiwata

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Correspondence to Shin'ichi Ishiwata.

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Uemura, S., Ishiwata, S. Loading direction regulates the affinity of ADP for kinesin. Nat Struct Mol Biol 10, 308–311 (2003). https://doi.org/10.1038/nsb911

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