Abstract
The effect of mechanical force on the free-energy surface that governs a chemical reaction is largely unknown. The combination of protein engineering with single-molecule force-clamp spectroscopy allows us to study the influence of mechanical force on the rate at which a protein disulfide bond is reduced by nucleophiles in a bimolecular substitution reaction (SN2). We found that cleavage of a protein disulfide bond by hydroxide anions exhibits an abrupt reactivity ‘switch’ at ∼500 pN, after which the accelerating effect of force on the rate of an SN2 chemical reaction greatly diminishes. We propose that an abrupt force-induced conformational change of the protein disulfide bond shifts its ground state, drastically changing its reactivity in SN2 chemical reactions. Our experiments directly demonstrate the action of a force-activated switch in the chemical reactivity of a single molecule.
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Acknowledgements
We thank L. Dougan, J. Alegre, P. Kosuri and other members of the Fernández laboratory for critical reading of the manuscript. S.G.-M. thanks the Generalitat de Catalunya for a postdoctoral fellowship through the NANO and Beatriu de Pinós programs, and also the Fundación Caja Madrid for financial support. This work was supported by NIH Grants (to J.M.F.).
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S.G.M. and J.M.F. conceived and designed the experiments; S.G.M., J.L., R.S. and T.K. performed the experiments; S.G.M., J.L., R.S. and T.K analysed the data; S.G.M. contributed materials and analysis tools; S.G.M and J.M.F. wrote the paper.
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Garcia-Manyes, S., Liang, J., Szoszkiewicz, R. et al. Force-activated reactivity switch in a bimolecular chemical reaction. Nature Chem 1, 236–242 (2009). https://doi.org/10.1038/nchem.207
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DOI: https://doi.org/10.1038/nchem.207
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