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Decoupling of nucleotide- and microtubule-binding sites in a kinesin mutant

Nature volume 396, pages 587–590 (1998)Cite this article

Abstract

Molecular motors require ATP to move along microtubules or actin filaments. To understand how molecular motors function, it is crucial to know how binding of the motor to its filamentous track stimulates the hydrolysis of ATP by the motor, enabling it to move along the filament. A mechanism for the enhanced ATP hydrolysis has not been elucidated, but it is generally accepted that conformational changes in the motor proteins1,2,3 occur when they bind to microtubules or actin filaments, facilitating the release of ADP. Here we report that a mutation in the motor domain of the microtubule motor proteins Kar3 and Ncd uncouples nucleotide- and microtubule-binding by the proteins, preventing activation of the motor ATPase by microtubules. Unlike the wild-type motors, the mutants bind tightly to both ADP and microtubules, indicating that interactions between the nucleotide- and microtubule-binding sites are blocked. The region of the motor that includes the mutated amino acid could transmit or undergo a conformational change required to convert the motor ATPase into a microtubule-stimulated state.

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Figure 1: Microtubule binding by wild-type Kar3 and Kar3-898.
Figure 2: ATP binding and hydrolysis by wild-type Kar3 and Kar3-898.
Figure 3: ATP hydrolysis and mant-ADP release by wild-type Kar3 and Kar3-898.
Figure 4: ATP hydrolysis and mant-ADP release by wild-type Ncd and Ncd-NK.

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Acknowledgements

We thank A. Hoyt for kar3-898, C. Fierke and K. Hightower for help with fluorometry and the use of a fluorometer, S. Rosenfeld for mant-ATP and A. Gulick for comments on the manuscript. This work was supported by the NIH.

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  1. Department of Microbiology, Duke University Medical Center, Durham, 27710, North Carolina, USA

    Hebok Song & Sharyn A. Endow

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Correspondence to Sharyn A. Endow.

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Song, H., Endow, S. Decoupling of nucleotide- and microtubule-binding sites in a kinesin mutant. Nature 396, 587–590 (1998). https://doi.org/10.1038/25153

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