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Protein Tethering for Single-Molecule Force Spectroscopy

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Single Molecule Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2694))

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Abstract

Molecular manipulation by optical tweezers is a central technique to study the folded states of individual proteins and how they depend on interactions with molecules including DNA, ligands, and other proteins. One of the key challenges of this approach is to stably attach DNA handles in an efficient manner. Here, we provide detailed descriptions of a universal approach to covalently link long DNA tethers of up to 5000 base pairs to proteins with or without native cysteines.

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Authors and Affiliations

  1. AMOLF, Amsterdam, The Netherlands

    Mario J. Avellaneda, Eline J. Koers, Vanda Sunderlikova & Sander J. Tans

Authors
  1. Mario J. Avellaneda
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  2. Eline J. Koers
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  3. Vanda Sunderlikova
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  4. Sander J. Tans
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Corresponding author

Correspondence to Sander J. Tans .

Editor information

Editors and Affiliations

  1. LaserLaB Amsterdam and Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands

    Iddo Heller

  2. LaserLaB Amsterdam and Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands

    David Dulin

  3. LaserLaB Amsterdam and Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands

    Erwin J.G. Peterman

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Avellaneda, M.J., Koers, E.J., Sunderlikova, V., Tans, S.J. (2024). Protein Tethering for Single-Molecule Force Spectroscopy. In: Heller, I., Dulin, D., Peterman, E.J. (eds) Single Molecule Analysis . Methods in Molecular Biology, vol 2694. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3377-9_3

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  • DOI: https://doi.org/10.1007/978-1-0716-3377-9_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3376-2

  • Online ISBN: 978-1-0716-3377-9

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