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Optical Tweezers-Based Measurements of Forces and Dynamics at Microtubule Ends

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Optical Tweezers

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1486))

Abstract

Microtubules are dynamic cytoskeletal polymers that polymerize and depolymerize while interacting with different proteins and structures within the cell. The highly regulated dynamic properties as well as the pushing and pulling forces generated by dynamic microtubule ends play important roles in processes such as in cell division. For instance, microtubule end-binding proteins are known to affect dramatically the dynamic properties of microtubules, and cortical dyneins are known to mediate pulling forces on microtubule ends. We discuss in this chapter our efforts to reconstitute these systems in vitro and mimic their interactions with structures within the cell using micro-fabricated barriers. Using an optical tweezers setup, we investigate the dynamics and forces of microtubules growing against functionalized barriers in the absence and presence of end-binding proteins and barrier-attached motor proteins. This setup allows high-speed as well as nanometer and piconewton resolution measurements on dynamic microtubules.

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Acknowledgments

We thank Matthew Footer for the help in setting up the axoneme purification from Tetrahymena thermophila and Samara Reck-Peterson and her lab for the training and assistance in setting up the dynein purification from Saccharomyces cerevisiae.

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Authors and Affiliations

  1. Department of Bionanoscience, Kavli Institute of NanoScience, Faculty of Applied Sciences, Delft University of Technology, Postbus 5046, 2600, GA, Delft, The Netherlands

    Marian Baclayon, Liedewij Laan & Marileen Dogterom

  2. FOM Institute AMOLF, Science Park 102, 1098 XG Amsterdam, Delft, The Netherlands

    Marian Baclayon, Svenja-Marei Kalisch, Ed Hendel, Liedewij Laan, Julien Husson, E. Laura Munteanu & Marileen Dogterom

Authors
  1. Marian Baclayon
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  2. Svenja-Marei Kalisch
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  3. Ed Hendel
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  4. Liedewij Laan
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  5. Julien Husson
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  6. E. Laura Munteanu
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  7. Marileen Dogterom
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Corresponding author

Correspondence to Marileen Dogterom .

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Editors and Affiliations

  1. Department of Anatomy and Structural Biology & Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, New York, USA

    Arne Gennerich

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Baclayon, M. et al. (2017). Optical Tweezers-Based Measurements of Forces and Dynamics at Microtubule Ends. In: Gennerich, A. (eds) Optical Tweezers. Methods in Molecular Biology, vol 1486. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6421-5_16

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  • DOI: https://doi.org/10.1007/978-1-4939-6421-5_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6419-2

  • Online ISBN: 978-1-4939-6421-5

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