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The Mitotic Spindle pp 47–61Cite as

Measuring the Effects of Microtubule-Associated Proteins on Microtubule Dynamics In Vitro

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1413))

Abstract

Microtubule dynamic instability, the process by which individual microtubules switch between phases of growth and shrinkage, is essential for establishing the architecture of cellular microtubule structures, such as the mitotic spindle. This switching process is regulated by a complex network of microtubule-associated proteins (MAPs), which modulate different aspects of microtubule dynamic behavior. To elucidate the effects of MAPs and their molecular mechanisms of action, in vitro reconstitution approaches with purified components are used. Here, I present methods for measuring individual and combined effects of MAPs on microtubule dynamics, using purified protein components and total-internal-reflection fluorescence (TIRF) microscopy. Particular focus is given to the experimental design, proper parameterization, and data analysis.

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Acknowledgements

I thank Anika Rahman for help with the data analysis. I am grateful to Justin Bois, Gary Brouhard, Melissa Gardner, Anneke Hibbel, Jonathon Howard, Elizabeth Lawrence, Chloe Snider, Michal Wieczorek, and especially Marija Podolski for helpful discussions and comments on the manuscript.

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Authors and Affiliations

  1. Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN, USA

    Marija Zanic

  2. Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN, USA

    Marija Zanic

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  1. Marija Zanic
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Corresponding author

Correspondence to Marija Zanic .

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Editors and Affiliations

  1. Massachusetts Institute of Technolo, Cambridge, Massachusetts, USA

    Paul Chang

  2. Dept of Cell & Developmental Biolog, Vanderbilt University, Nashville, Tennessee, USA

    Ryoma Ohi

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Zanic, M. (2016). Measuring the Effects of Microtubule-Associated Proteins on Microtubule Dynamics In Vitro. In: Chang, P., Ohi, R. (eds) The Mitotic Spindle. Methods in Molecular Biology, vol 1413. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3542-0_4

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  • DOI: https://doi.org/10.1007/978-1-4939-3542-0_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3540-6

  • Online ISBN: 978-1-4939-3542-0

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