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In Vitro Microtubule Severing Assays

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Adhesion Protein Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1046))

Abstract

Microtubules are rigid and highly dynamic cellular polymers essential for intracellular transport, cell division and differentiation. Their stability is tightly regulated by a vast array of cellular factors. In vitro microtubule assays have proven to be powerful tools for deciphering the mechanism of microtubule dynamics regulators such as molecular motors and microtubule associated proteins. In this chapter we focus on microtubule severing enzymes that use the energy of ATP hydrolysis to introduce internal breaks in the microtubule lattice. We present a detailed protocol for a light microscopy based in vitro microtubule severing assay that was instrumental in the identification and characterization of these enzymes.

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Acknowledgments

A. Roll-Mecak is supported by the National Institutes of Health Intramural Program. N. E. Ziółkowska is supported by a Searle Scholar Award to A. Roll-Mecak.

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Authors and Affiliations

  1. Cell Biology and Biophysics Unit, National Institute of Neurological Disorders and Stroke, Bethesda, MD, USA

    Natasza E. Ziółkowska

  2. Cell Biology and Biophysics Unit, Center for Biophysics, National Institute of Neurological Disorders and Stroke, National Heart, Lung and Blood Institute, Bethesda, MD, USA

    Antonina Roll-Mecak

Authors
  1. Natasza E. Ziółkowska
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  2. Antonina Roll-Mecak
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Editor information

Editors and Affiliations

  1. , Department of Oncology, University of Oxford, Roosevelt Drive, Oxford, OX3 7DQ, United Kingdom

    Amanda S. Coutts

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Ziółkowska, N.E., Roll-Mecak, A. (2013). In Vitro Microtubule Severing Assays. In: Coutts, A. (eds) Adhesion Protein Protocols. Methods in Molecular Biology, vol 1046. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-538-5_19

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  • DOI: https://doi.org/10.1007/978-1-62703-538-5_19

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-537-8

  • Online ISBN: 978-1-62703-538-5

  • eBook Packages: Springer Protocols

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