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Two conformations in the human kinesin power stroke defined by X-ray crystallography and EPR spectroscopy

Nature Structural Biology volume 9, pages 844–848 (2002)Cite this article

Abstract

Crystal structures of the molecular motor kinesin show conformational variability in a structural element called the neck linker. Conformational change in the neck linker, initiated by ATP exchange, is thought to drive the movement of kinesin along the microtubule track. We use site-specific EPR measurements to show that when microtubules are absent, the neck linker exists in equilibrium between two structural states (disordered and 'docked'). The active site nucleotide does not control the position taken by the neck linker. However, we find that sulfate can specifically bind near the nucleotide site and stabilize the docked neck linker conformation, which we confirmed by solving a new crystal structure. Comparing the crystal structures of our construct with the docked or undocked neck linker reveals how microtubule binding may activate the nucleotide-sensing mechanism of kinesin, allowing neck linker transitions to power motility.

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Figure 1: Two crystallized conformations of human kinesin.
Figure 2: EPR measurements of site-labeled probes on the neck linker.
Figure 3: A model for microtubule-induced coupling of switch II.
Figure 4: An ion-binding site in K349.

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Acknowledgements

We are indebted to E. Sablin, who provided many insightful comments during the preparation of the manuscript. We also give our thanks to V. Minor for enthusiastic help in the indexing and scaling of diffraction data, and P. Fajer for helpful comments on the EPR analysis. The National Institutes of Health supported this work.

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Authors and Affiliations

  1. Department of Biochemistry and Biophysics, University of California, San Francisco, 94143, California, USA

    Charles V. Sindelar, Mary Jane Budny, Nariman Naber, Robert Fletterick & Roger Cooke

  2. Department of Biochemistry, Stanford University, Palo Alto, 94305, California, USA

    Sarah Rice

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  1. Charles V. Sindelar
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  3. Sarah Rice
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Correspondence to Robert Fletterick.

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Sindelar, C., Budny, M., Rice, S. et al. Two conformations in the human kinesin power stroke defined by X-ray crystallography and EPR spectroscopy. Nat Struct Mol Biol 9, 844–848 (2002). https://doi.org/10.1038/nsb852

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