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Stabilization of Proteins by Freeze-Drying in the Presence of Trehalose: A Case Study of Tubulin

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Book cover Protein Downstream Processing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2178))

Abstract

Microtubules, polymers of the heterodimeric protein αβ-tubulin, are indispensable for many cellular activities such as maintenance of cell shape, division, migration, and ordered vesicle transport. In vitro assays to study microtubule functions and their regulation by associated proteins require the availability of assembly-competent purified tubulin. However, tubulin is a thermolabile protein that rapidly converts into a nonpolymerizing state. For this reason, it is usually stored at −80 °C or liquid nitrogen to preserve its conformation and polymerization properties. In this chapter, we describe a method for freeze-drying of assembly-competent tubulin in the presence of nonreducing sugar trehalose, and methods enabling the evaluation of tubulin functions in rehydrated samples.

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Acknowledgments

This work was supported in part by Grants 18-27197S and 19-20716S from the Czech Science Foundation, Grant LTAUSA17052 from Ministry of Education, Youth, and Sports of the Czech Republic, Project NAS-17-11 from Academy of Sciences of the Czech Republic, and by the Institutional Research Support (RVO 68378050).

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Authors and Affiliations

  1. Laboratory of Biology of Cytoskeleton, Institute of Molecular Genetics of the Czech Academy of Sciences, Prague, Czech Republic

    Pavel Dráber, Vadym Sulimenko, Tetyana Sulimenko & Eduarda Dráberová

Authors
  1. Pavel Dráber
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  2. Vadym Sulimenko
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  3. Tetyana Sulimenko
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  4. Eduarda Dráberová
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Corresponding author

Correspondence to Pavel Dráber .

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Editors and Affiliations

  1. Laboratory of Enzyme Technology, Department of Biotechnology, School of Applied Biology and Biotechnology, Agricultural University of Athens, Athens, Greece

    Nikolaos E. Labrou

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Dráber, P., Sulimenko, V., Sulimenko, T., Dráberová, E. (2021). Stabilization of Proteins by Freeze-Drying in the Presence of Trehalose: A Case Study of Tubulin. In: Labrou, N.E. (eds) Protein Downstream Processing. Methods in Molecular Biology, vol 2178. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0775-6_27

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  • DOI: https://doi.org/10.1007/978-1-0716-0775-6_27

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0774-9

  • Online ISBN: 978-1-0716-0775-6

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