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F 1 F 2-selective NMR spectroscopy

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Abstract

Fourier transform NMR spectroscopy has provided unprecedented insight into the structure, interaction and dynamic motion of proteins and nucleic acids. Conventional biomolecular NMR relies on the acquisition of three-dimensional and four-dimensional (4D) data matrices to establish correlations between chemical shifts in the frequency domains F 1, F 2, F 3 and F 1, F 2, F 3, F 4 respectively. While rich in information, these datasets require a substantial amount of acquisition time, are visually highly unintuitive, require expert knowledge to process, and sample dark and bright regions of the frequency domains equally. Here, we present an alternative approach to obtain multidimensional chemical shift correlations for biomolecules. This strategy focuses on one narrow frequency range, F 1 F 2, at a time and records the resulting F 3 F 4 correlation spectrum by two-dimensional NMR. As a result, only regions of the frequency domain that contain signals in F 1 F 2 (“bright regions”) are sampled. F 1 F 2 selection is achieved by Hartmann–Hahn cross-polarization using weak radio frequency fields. This approach reveals information equivalent to that of a conventional 4D experiment, while the dimensional reduction may shorten the total acquisition time and simplifies spectral processing, interpretation and comparative analysis. Potential applicability of the F 1 F 2-selective approach is illustrated by de novo assignment, structural and dynamics studies of ubiquitin and fatty-acid binding protein 4 (FABP4). Further extension of this concept may spawn new selective NMR experiments to aid studies of site-specific structural dynamics, protein–protein interactions and allosteric modulation of protein structure.

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Abbreviations

CP:

Cross-polarization

FID:

Free induction decay

FT:

Fourier transform

HSQC:

Heteronuclear single-quantum coherence

NOE:

Nuclear overhauser effect

NUS:

Non-uniform sampling

PEP:

Preservation of equivalent pathways

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Authors and Affiliations

  1. Department of Molecular and Cellular Physiology, Graduate School of Medicine, Kyoto University, Sakyo-ku Yoshida Konoe-cho, Kyoto, 606-8501, Japan

    Erik Walinda

  2. Department of Molecular Engineering, Graduate School of Engineering, Kyoto University, Nishikyo-ku Kyoto-Daigaku Katsura, Kyoto, 615-8510, Japan

    Daichi Morimoto, Masahiro Shirakawa & Kenji Sugase

Authors
  1. Erik Walinda
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  2. Daichi Morimoto
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  3. Masahiro Shirakawa
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  4. Kenji Sugase
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Corresponding author

Correspondence to Kenji Sugase.

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Walinda, E., Morimoto, D., Shirakawa, M. et al. F 1 F 2-selective NMR spectroscopy. J Biomol NMR 68, 41–52 (2017). https://doi.org/10.1007/s10858-017-0113-x

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