Journal of Biomolecular NMR

, Volume 68, Issue 1, pp 41–52 | Cite as

F 1 F 2-selective NMR spectroscopy

  • Erik Walinda
  • Daichi Morimoto
  • Masahiro Shirakawa
  • Kenji SugaseEmail author


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.


Frequency selection Dimensional reduction Cross-polarization Selective excitation Nuclear overhauser effect spectroscopy Relaxation dispersion 





Free induction decay


Fourier transform


Heteronuclear single-quantum coherence


Nuclear overhauser effect


Non-uniform sampling


Preservation of equivalent pathways

Supplementary material

10858_2017_113_MOESM1_ESM.docx (881 kb)
Supplementary material 1 (DOCX 880 KB)


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Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  • Erik Walinda
    • 1
  • Daichi Morimoto
    • 2
  • Masahiro Shirakawa
    • 2
  • Kenji Sugase
    • 2
    Email author
  1. 1.Department of Molecular and Cellular Physiology, Graduate School of MedicineKyoto UniversityKyotoJapan
  2. 2.Department of Molecular Engineering, Graduate School of EngineeringKyoto UniversityKyotoJapan

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