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Kinesin hydrolyses one ATP per 8-nm step

Nature volume 388, pages 386–390 (1997)Cite this article

Abstract

Kinesin is a two-headed, ATP-dependent motor protein1,2 that moves along microtubules indiscrete steps3 of 8 nm. In vitro, single molecules produceprocessive movement4,5, motors typically take ∼100steps before releasing from a microtubule5,6,7 . A central question relates tomechanochemical coupling in this enzyme: how many molecules ofATP are consumed per step? For the actomyosin system,experimental approaches to this issue have generated considerablecontroversy8,9. Here we take advantage of theprocessivity of kinesin to determine the coupling ratio withoutrecourse to direct measurements of ATPase activity, which aresubject to large experimental uncertainties8,10,11,12. Beads carrying singlemolecules of kinesin moving on microtubules were tracked with highspatial and temporal resolution by interferometry3,13. Statistical analysis of theintervals between steps at limiting ATP, and studies offluctuations in motor speed as a function of ATPconcentration14,15, allow the coupling ratio to bedetermined. At near-zero load, kinesin moleculeshydrolyse a single ATP molecule per 8-nm advance. Thisfinding excludes various one-to-many andmany-to-one coupling schemes, analogous to thoseadvanced for myosin, and places severe constraints on models for movement.

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Figure 1: Average bead velocity, v, versus ATP concentration (double logarithmic plot).
Figure 2: a, Sample record of movement at 2 µM ATP, showing the elementary steps (solid line).
Figure 3: a, Histogram of times between steps for records at limiting ATP concentration (0.75, 1 and 2 µM).

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Acknowledgements

We thank S. Gross, W. Ryu, L. Satterwhite, M. Wang and especially K. Visscher for technical assistance and discussions; S. Gross for help in purifying kinesin; and P. Mitra and K. Svoboda for advice in the early stages of this project. This work was supported by a grant from NIGMS (S.M.B.) and predoctoral fellowships from NSF and American Heart Association (M.J.S.).

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Authors and Affiliations

  1. *Departments of Physics, Princeton University, Princeton, 08544, New Jersey, USA

    Mark J. Schnitzer

  2. †Molecular Biology, Princeton University , Princeton, 08544, New Jersey, USA

    Mark J. Schnitzer & Steven M. Block

  3. ‡Princeton Materials Institute, Princeton University, Princeton, 08544, New Jersey, USA

    Steven M. Block

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Correspondence to Mark J. Schnitzer.

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Schnitzer, M., Block, S. Kinesin hydrolyses one ATP per 8-nm step. Nature 388, 386–390 (1997). https://doi.org/10.1038/41111

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