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Quantum-dot-assisted characterization of microtubule rotations during cargo transport

Nature Nanotechnology volume 3, pages 552–556 (2008)Cite this article

Abstract

Owing to their wide spectrum of in vivo functions, motor proteins, such as kinesin-1, show great potential for application as nanomachines in engineered environments. When attached to a substrate surface, these motors are envisioned to shuttle cargo that is bound to reconstituted microtubules—one component of the cell cytoskeleton—from one location to another1,2. One potentially serious problem for such applications is, however, the rotation of the microtubules around their longitudinal axis3,4. Here we explore this issue by labelling the gliding microtubules with quantum dots to simultaneously follow their sinusoidal side-to-side and up-and-down motion in three dimensions with nanometre accuracy. Microtubule rotation, which originates from the kinesin moving along the individual protofilaments of the microtubule, was not impeded by the quantum dots. However, pick-up of large cargo inhibited the rotation but did not affect the velocity of microtubule gliding. Our data show that kinesin-driven microtubules make flexible, responsive and effective molecular shuttles for nanotransport applications.

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Figure 1: Principle of QD-assisted 3D imaging of microtubule rotation.
Figure 2: FLIC microscopy of QDs attached to microtubules driven by kinesin-1.
Figure 3: Determining the direction of rotation using dual-colour FLIC imaging.
Figure 4: Microtubule rotations during pick-up and drop-off of micrometre-sized cargo.

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Acknowledgements

The authors thank C. Bräuer, D. Naumburger and F. Friedrich for technical support, D. Zwicker and Y. Kalaidzidis for help with the tracking algorithms, G. Brouhard for advice on the subtilisin digestion of microtubules, C. Leduc, J. Kerssemakers, J. Helenius, S. Mashaghi and J. Howard for fruitful discussions, and C. Gell and T. Korten for comments on the manuscript. This work was supported by the German Federal Ministry of Education and Research (grant 03 N 8712) and the Max-Planck Society.

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  1. Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, 01307, Germany

    Bert Nitzsche, Felix Ruhnow & Stefan Diez

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  1. Bert Nitzsche
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  2. Felix Ruhnow
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  3. Stefan Diez
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Contributions

B.N. and S.D. conceived and designed the experiments. B.N. performed the experiments. F.R. developed the nanometre tracking software. B.N. and F.R. analysed the data. All authors discussed the results. B.N. and S.D. co-wrote the paper.

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Correspondence to Stefan Diez.

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Nitzsche, B., Ruhnow, F. & Diez, S. Quantum-dot-assisted characterization of microtubule rotations during cargo transport. Nature Nanotech 3, 552–556 (2008). https://doi.org/10.1038/nnano.2008.216

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