TROSY pulse sequence for simultaneous measurement of the 15N R1 and {1H}–15N NOE in deuterated proteins

Communication
  • 42 Downloads

Abstract

A TROSY-based NMR experiment is described for simultaneous measurement of the 15N longitudinal relaxation rate constant R1 and the {1H}–15N nuclear Overhauser enhancement. The experiment is based on the observation that the TROSY mixing pulse sequence element symmetrically exchanges 1H and 15N magnetizations. The accuracy of the proposed technique is validated by comparison to independent measurements of both relaxation parameters for the protein ubiquitin. The simultaneous experiment is approximately 20–33% shorter than conventional sequential measurements.

Keywords

Dynamics Longitudinal relaxation Nuclear Overhauser enhancement Protein Spin–lattice relaxation TROSY 

Notes

Acknowledgements

Support from National Institutes of Health Grants R01 GM050291 (A.G.P.) and T32 GM008281 (P.A.O.) is acknowledged gratefully. We thank Mark Rance (University of Cincinnati) for helpful discussions. Some of the work presented here was conducted at the Center on Macromolecular Dynamics by NMR Spectroscopy located at the New York Structural Biology Center, supported by a Grant from the NIH National Institute of General Medical Sciences (P41 GM118302). A.G.P. is a member of the New York Structural Biology Center.

Supplementary material

10858_2018_181_MOESM1_ESM.txt (5 kb)
Supplementary material 1 (TXT 5 KB)
10858_2018_181_MOESM2_ESM.txt (9 kb)
Supplementary material 2 (TXT 8 KB)
10858_2018_181_MOESM3_ESM.csv (6 kb)
Supplementary material 3 (CSV 6 KB)

References

  1. Abyzov A et al (2016) Identification of dynamic modes in an intrinsically disordered protein using temperature-dependent NMR relaxation. J Am Chem Soc 138:6240–6251CrossRefGoogle Scholar
  2. Cavanagh J, Fairbrother WJ, Palmer AG III, Skelton NJ (2007) Protein NMR spectroscopy, 2nd edn. Academic Press, Burlington, pp 679–724CrossRefGoogle Scholar
  3. Delaglio F et al (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293CrossRefGoogle Scholar
  4. Favier A, Brutscher B (2011) Recovering lost magnetization: polarization enhancement in biomolecular NMR. J Biomol NMR 49:9–15CrossRefGoogle Scholar
  5. Ferrage F, Cowburn D, Ghose R (2009) Accurate sampling of high-frequency motions in proteins by steady-state 15N-{1H} nuclear Overhauser effect measurements in the presence of cross-correlated relaxation. J Am Chem Soc 131:6048–6049CrossRefGoogle Scholar
  6. Findeisen M, Brand T, Berger S (2007) A 1H-NMR thermometer suitable for cryoprobes. Magn Reson Chem 45:175–178CrossRefGoogle Scholar
  7. Gairi M et al (2015) An optimized method for 15N R1 relaxation rate measurements in non-deuterated proteins. J Biomol NMR 62:209–220CrossRefGoogle Scholar
  8. Gill ML, Byrd RA, Palmer AG (2016) Dynamics of GCN4 facilitate DNA interaction: a model-free analysis of an intrinsically disordered region. Phys Chem Chem Phys 18:5839–5849CrossRefGoogle Scholar
  9. Hsu A, O’Brien PA, Bhattacharya S, Rance M, Palmer AG (2017) Enhanced spectral density mapping through combined multiple-field deuterium 13CH2D methyl spin relaxation NMR spectroscopy. Methods.  https://doi.org/10.1016/j.ymeth.2017.12.020 Google Scholar
  10. Lakomek NA, Ying J, Bax A (2012) Measurement of 15N relaxation rates in perdeuterated proteins by TROSY-based methods. J Biomol NMR 53:209–221CrossRefGoogle Scholar
  11. Palmer AG (2004) NMR characterization of the dynamics of biomacromolecules. Chem Rev 104:3623–3640CrossRefGoogle Scholar
  12. Peng JW, Wagner G (1995) Frequency spectrum of NH bonds in eglin c from spectral density mapping at multiple fields. Biochemistry 34:16733–16752CrossRefGoogle Scholar
  13. Zhu G, Xia Y, Nicholson LK, Sze KH (2000) Protein dynamics measurements by TROSY-based NMR experiments. J Magn Reson 143:423–426ADSCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular BiophysicsColumbia UniversityNew YorkUSA

Personalised recommendations