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A primary hydrogen–deuterium isotope effect observed at the single-molecule level

Nature Chemistry volume 2, pages 921–928 (2010)Cite this article

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Abstract

The covalent chemistry of reactants tethered within a single protein pore can be monitored by observing the time-dependence of ionic current flow through the pore, which responds to bond making and breaking in individual reactant molecules. Here we use this ‘nanoreactor’ approach to examine the reaction of a quinone with a thiol to form a substituted hydroquinone by reductive 1,4-Michael addition. Remarkably, a primary hydrogen–deuterium isotope effect is readily detected at the single-molecule level during prototropic rearrangement of an initial adduct. The observation of individual reaction intermediates allows the measurement of an isotope effect whether or not the step involved is rate limiting, which would not be the case in an ensemble measurement.

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Figure 1: Single-molecule quinone chemistry.
Figure 2: Single-molecule reactions of UQ (1) and UQ(D) (1) with the thiol of PSH at pH 6.5.
Figure 3: Single-molecule reactions of UQ (1) and UQ(D) (1) with the thiol of PSH at pH 4.5.
Figure 4: Reaction of UQ (1) and UQ(D) (1) with glutathione in bulk solution.
Figure 5: QuB simulations of the reaction of UQ (1) and UQ(D) (1) with the thiol of PSH.

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Acknowledgements

This work was supported by the MRC.

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  1. Siran Lu and Wen-Wu Li: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Chemistry, University of Oxford, Oxford, OX1 3TA, UK

    Siran Lu, Wen-Wu Li, Dvir Rotem, Ellina Mikhailova & Hagan Bayley

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Contributions

S.L. and W-W.L. performed the kinetic measurements and analysed the data. W-W.L. synthesized deuterated UQ (1). S.L. and E.M. made the PSH pore. S.L, W-W.L. and D.R. performed simulations using QuB. S.L., W-W.L. and H.B. planned the research and wrote the paper.

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Correspondence to Hagan Bayley.

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Lu, S., Li, WW., Rotem, D. et al. A primary hydrogen–deuterium isotope effect observed at the single-molecule level. Nature Chem 2, 921–928 (2010). https://doi.org/10.1038/nchem.821

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