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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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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