Abstract
The nuclear spin-lattice relaxation rate 1/T1 depends on the correlation time τc of the molecule bearing the nuclear spin, and can therefore probe changes of τc upon binding of a rapidly moving small ligand to a more slowly moving larger protein. In practice however, the dependence is such that only a small difference in relaxation rate is obtained at high field. Here we present a scheme in which nuclear spins are first hyperpolarized using DNP, and then allowed to relax at low magnetic field in presence of a target protein, which generates a large T1 contrast. The sample is subsequently transferred into a conventional nuclear magnetic resonance probe (NMR), where the effect of the low-field relaxation is read out using high-field liquid-state NMR. Using only 14 μM of a 13C-labeled reporter ligand, we observe protein binding reliably for protein concentrations as low as 2 μM in a single scan. The scheme is expanded to a label-free ligand via a competitive binding experiment in which the label-free ligand displaces the 13C-labeled reporter ligand.
Data availability
The NMR raw data and processing files are available from KITopen at https://doi.org/10.35097/4df4g6du1awng50h (ref. 36).
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Acknowledgements
This work has received funding from the European Research Council (ERC) via the Synergy grant “Highly Informative Drug Screening by Overcoming NMR Restrictions” (HiSCORE, grant agreement no. 951459), from the “Impuls- und Vernetzungsfonds of the Helmholtz-Association” (grant number VH-NG-1432), and from the Deutsche Forschungsgemeinschaft (DFG, grant number 454252029 - SFB1527).
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P.N. conducted the experiment with support from M.M. and N.L. P.N. analyzed data with support from B.M. A.G., B.M., and P.N. conceived the experiment. N.L. produced the PHD protein, and N.L., A.G., and P.N. wrote and adapted the pulse sequence. B.M. and P.N. wrote the manuscript with contributions by A.G. and N.L. All authors reviewed the manuscript.
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All authors have filed an EU patent application on drug screening using DNP and low-field relaxation as a contrast mechanism (EP 25152466.6). B.M. is co-owner of HyperSpin Scientific UG.
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Communications Chemistry thanks Samuel F. Cousin and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. A peer review file is available.
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Narwal, P., Lorz, N., Minaei, M. et al. Single-scan detection of ligand-binding using hyperpolarization and low-field relaxation. Commun Chem (2026). https://doi.org/10.1038/s42004-026-01934-7
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DOI: https://doi.org/10.1038/s42004-026-01934-7
