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Journal Articles Methods in Molecular Biology Year : 2018

High-Speed Force Spectroscopy for Single Protein Unfolding

Abstract

Single-molecule force spectroscopy (SMFS) measurements allow for quantification of the molecular forces required to unfold individual protein domains. Atomic force microscopy (AFM) is one of the long-established techniques for force spectroscopy (FS). Although FS at conventional AFM pulling rates provides valuable information on protein unfolding, in order to get a more complete picture of the mechanism, explore new regimes, and combine and compare experiments with simulations, we need higher pulling rates and μs-time resolution, now accessible via high-speed force spectroscopy (HS-FS). In this chapter, we provide a step-by-step protocol of HS-FS including sample preparation, measurements and analysis of the acquired data using HS-AFM with an illustrative example on unfolding of a well-studied concatamer made of eight repeats of the titin I91 domain.
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Dates and versions

hal-01838371 , version 1 (05-10-2018)

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Fidan Sumbul, Arin Marchesi, Hirohide Takahashi, Simon Scheuring, Felix Rico. High-Speed Force Spectroscopy for Single Protein Unfolding. Methods in Molecular Biology, 2018, pp.243-264. ⟨10.1007/978-1-4939-8591-3_15⟩. ⟨hal-01838371⟩
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