Abstract
Microtubules are essential cellular structures in plant cells. They are polymerized from tubulin dimers and are regulated by microtubule-associated proteins (MAPs). Here, we describe a protocol for purifying tubulin dimers and MAPs from plant cells. The protocol involves preparing vacuole-free mini-protoplasts, a high quality cytoplasmic extract, cycles of microtubule polymerization and depolymerization to increase tubulin and MAP concentration, separation of tubulin and MAPs by column chromatography. We also present tubulin purification methods for biochemical assays.
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Acknowledgements
We are grateful to Prof. Tobias Baskin (University of Massachusetts, Amherst) for helpful discussion and critical editing of the manuscript. We also thank Prof. Bo Liu (University of California, Davis) for helpful comments to the manuscript. This work was supported by MEXT/JSPS KAKENHI Grant Number 15H05598 and 16H01229.
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Hamada, T., Sonobe, S. (2017). Isolation of Microtubules and Microtubule-Associated Proteins. In: Taylor, N., Millar, A. (eds) Isolation of Plant Organelles and Structures. Methods in Molecular Biology, vol 1511. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6533-5_22
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DOI: https://doi.org/10.1007/978-1-4939-6533-5_22
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