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Unfolding and Refolding Proteins Using Single-Molecule AFM

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Single Molecule Analysis

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

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Abstract

Single-molecule atomic force microscopy (AFM) allows capturing the conformational dynamics of an individual molecule under force. In this chapter, we describe a protocol for conducting a protein nanomechanical experiment using AFM, covering both the force-extension and force-clamp modes. Combined, these experiments provide an integrated vista of the molecular mechanisms—and their associated kinetics—underpinning the mechanical unfolding and refolding of individual proteins when exposed to mechanical load.

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Acknowledgments

R.T-R. is recipient of a King’s Prize Fellowship. This work is supported by the European Commission (Mechanocontrol, Grant Agreement 731957), BBSRC sLoLa (B/V003518/1), Leverhulme Trust Research Leadership Award (RL 2016-015), Wellcome Trust Investigator Award (212218/Z/18/Z), and Royal Society Wolfson Fellowship (RSWF/R3/183006) all to S.G.M.

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Authors and Affiliations

  1. Department of Physics, Randall Centre for Cell and Molecular Biophysics and London Centre for Nanotechnology, King’s College London, London, UK

    Marc Mora, Rafael Tapia-Rojo & Sergi Garcia-Manyes

Authors
  1. Marc Mora
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  2. Rafael Tapia-Rojo
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  3. Sergi Garcia-Manyes
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Corresponding author

Correspondence to Sergi Garcia-Manyes .

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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Mora, M., Tapia-Rojo, R., Garcia-Manyes, S. (2024). Unfolding and Refolding Proteins Using Single-Molecule AFM. 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_16

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  • DOI: https://doi.org/10.1007/978-1-0716-3377-9_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3376-2

  • Online ISBN: 978-1-0716-3377-9

  • eBook Packages: Springer Protocols

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