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  • Review Article
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Ensemble determination by NMR data deconvolution

Nature Reviews Chemistry volume 7, pages 511–524 (2023)Cite this article

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Abstract

Nuclear magnetic resonance (NMR) is the spectroscopic technique of choice for determining molecular conformations in solution at atomic resolution. As solution NMR spectra are rich in structural and dynamic information, the way in which the data should be acquired and handled to deliver accurate ensembles is not trivial. This Review provides a guide to the NMR experiment selection and parametrization process, the generation of viable theoretical conformer pools and the deconvolution of time-averaged NMR data into a conformer ensemble that accurately represents a flexible molecule in solution. In addition to reviewing the key elements of solution ensemble determination of flexible mid-sized molecules, the feasibility and pitfalls of data deconvolution are discussed with a comparison of the performance of representative algorithms.

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Fig. 1: Workflow of NMR data deconvolution.
Fig. 2: Nonlinear scaling of nuclear Overhauser effect-derived interproton distances.
Fig. 3: Nuclear Overhauser effect build-up curves.
Fig. 4: Common errors observed in nuclear Overhauser effect build-up curves.
Fig. 5: Maximum nuclear Overhauser effect enhancement.
Fig. 6: Compounds used for method validation and comparison.

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Data availability

The original NMR spectroscopic (FIDs) and computational data (structure files); input and output files for NAMFIS, NMR_FIT, DISCON and StereoFitter; the simulated and experimental data for validation of the deconvolution algorithms; and the installation files, along with instructions for the installation of NAMFIS, are available at Zenodo: https://doi.org/10.5281/zenodo.6866837.

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Acknowledgements

The authors thank S. Wlodek (OpenEye) for giving us early access to NMR_FIT, O. Atasoylu for his help with DISCON, and P. Monje and A. Navarro-Vazquez for their guidance with StereoFitter. The authors are grateful to J. Bogaerts and A. Demkiv for their Linux support.

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Authors and Affiliations

  1. Department of Chemistry—BMC, Uppsala University, Uppsala, Sweden

    Lianne H. E. Wieske, Stefan Peintner & Máté Erdélyi

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  1. Lianne H. E. Wieske
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Supplementary information

Glossary

Initial-rate approximation

The assumption that the nuclear Overhauser effect observed at short mixing times, in the linear regime of the build-up curve, is proportional to the interproton distance.

Longitudinal relaxation

The time needed for a given spin to fall back to equilibrium. Also known as spin–lattice relaxation.

Mixing time

The time delay in the nuclear Overhauser effect (NOE) spectroscopy pulse sequence during which cross-relaxation occurs, giving rise to the NOE. Within the linear build-up regime, the longer the mixing time, the more time is given for the magnetization to transfer between spins, resulting in stronger NOE cross peaks.

NOE build-up curves

Series of nuclear Overhauser effect (NOE) experiments with at least four mixing times recorded to determine the NOE build-up rate, which is the rate of NOE signal evolution. At short enough mixing times, the intensity of the NOE cross peaks linearly correlates to the mixing time, and the slope of the signal build-ups provides population-averaged interproton distances.

Non-uniform sampling

A way of recording 2D NMR spectra whereby only a subset of increments (number of points in the f1 dimension) is collected, and the missing increments need to be reconstructed before Fourier transform.

Nuclear Overhauser effect

(NOE). A through-space dipole–dipole interaction between spins; the magnitude of the effect depends on the distance separation of the spins, among other factors.

Rotating-frame Overhauser effect spectroscopy

(ROESY). A variation of the nuclear Overhauser effect spectroscopy sequence with the difference that the signals of a ROESY are always positive, independent of the correlation time and spectrometer frequency.

Solution ensembles

Collection of different conformations a molecule adopts in solution.

Solvent suppression

A pulse or pulse combination used with the aim of removing a strong solvent signal from a spectrum, to avoid overloading of the analogue-to-digital converter (ADC).

t 1 noise

Noise visible in 2D spectra along the indirect (f1) axis owing to too few increments, resulting in a truncated free induction delay, and after Fourier transformation, manifesting as wiggles in the baseline; most commonly seen at intense singlets within a spectrum. To a certain extent, it can be compensated with weighting functions.

T 2 relaxation

Process by which the transverse components of magnetization dephase, causing the signal intensity to exponentially decay. The time constant T2 is the time required for the transverse magnetization to fall to ~37% (1/e) of its initial value.

Total correlation spectroscopy

(TOCSY). A homonuclear experiment that allows for the detection of covalently connected spins but is not restricted to the closest neighbours.

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Wieske, L.H.E., Peintner, S. & Erdélyi, M. Ensemble determination by NMR data deconvolution. Nat Rev Chem 7, 511–524 (2023). https://doi.org/10.1038/s41570-023-00494-x

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