Abstract
Kinesin is an ATP-driven molecular motor protein that moves processively along microtubules. Despite considerable research, the detailed mechanism of kinesin motion remains elusive. We applied an enhanced suite of single- and multiple-molecule fluorescence polarization microscopy assays to report the orientation and mobility of kinesin molecules bound to microtubules as a function of nucleotide state. In the presence of analogs of ATP, ADP-Pi or in the absence of nucleotide, the kinesin head maintains a rigid orientation. In the presence of ADP, the motor domain of kinesin, still bound to the microtubule, adopts a previously undescribed, highly mobile state. This state may be general to the chemomechanical cycle of motor proteins; in the case of kinesin, the transition from a highly mobile to a rigid state after ADP release may contribute to the generation of the 8 nm step.
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Acknowledgements
We thank R. Sakowicz for axoneme preparations, advice and discussions; R. Dickson, R. Vale and D. Pierce for their advice in the initial phases of this project; L. Gross, H. Deng and L. Siconolfi-Baez for mass spectrometry analysis; A. Asenjo for biochemical assays, and S. Brasselet and B. Lounis for helpful discussions and experimental suggestions. L.S.B. Goldstein is an investigator of the Howard Hughes Medical Institute. This project was supported by NSF grants.
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Sosa, H., Peterman, E., Moerner, W. et al. ADP-induced rocking of the kinesin motor domain revealed by single-molecule fluorescence polarization microscopy. Nat Struct Mol Biol 8, 540–544 (2001). https://doi.org/10.1038/88611
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DOI: https://doi.org/10.1038/88611
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