Journal of Biomolecular NMR

, Volume 67, Issue 3, pp 233–241 | Cite as

NOESY-WaterControl: a new NOESY sequence for the observation of under-water protein resonances

  • Allan M. Torres
  • Gang ZhengEmail author
  • William S. Price


Highly selective and efficient water signal suppression is indispensable in biomolecular 2D nuclear Overhauser effect spectroscopy (NOESY) experiments. However, the application of conventional water suppression schemes can cause a significant or complete loss of the biomolecular resonances at and around the water chemical shift (ω2). In this study, a new sequence, NOESY-WaterControl, was developed to address this issue. The new sequence was tested on lysozyme and bovine pancreatic trypsin inhibitor (BPTI), demonstrating its efficiency in both water suppression and, more excitingly, preserving water-proximate biomolecular resonances in ω2. The 2D NOESY maps obtained using the new sequence thus provide more information than the maps obtained with conventional water suppression, thereby lessening the number of experiments needed to complete resonance assignments of biomolecules. The 2D NOESY-WaterControl map of BPTI showed strong bound water and exchangeable proton signals in ω1 but these signals were absent in ω2, indicating the possibility of using the new sequence to discriminate bound water and exchangeable proton resonances from non-labile proton resonances with similar chemical shifts to water.


Biomolecule Diffusion NMR NOESY PGSE Protein NMR Solvent signal suppression Stimulated echo 

Supplementary material

10858_2017_100_MOESM1_ESM.doc (56 kb)
Supplementary material 1 (DOC 56 KB)


  1. Brown SC, Weber PL, Mueller L (1988) Toward complete 1H NMR spectra in proteins. J Magn Reson 77:166–169ADSGoogle Scholar
  2. Chakrabarti G, Kim S, Gupta ML, Barton JS, Himes RH (1999) Stabilization of tubulin by deuterium oxide. Biochemistry 38:3067–3072CrossRefGoogle Scholar
  3. Hoult DI (1976) Solvent peak saturation with single phase and quadrature Fourier transformation. J Magn Reson 21:337–347ADSGoogle Scholar
  4. Hoult DI, Richards RE (1975) Critical factors in the design of sensitive high resolution nuclear magnetic resonance spectrometers. Proc R Soc Lond A 344:311–340ADSCrossRefGoogle Scholar
  5. Hwang TL, Shaka AJ (1995) Water suppression that works. Excitation sculpting using arbitrary wave-forms and pulsed-field gradients. J Magn Reson 112A:275–279ADSCrossRefGoogle Scholar
  6. Krishna NR (1976) A method for solvent peak suppression in FTNMR spectra by double resonance. J Magn Reson 22:555–559ADSGoogle Scholar
  7. Kumar A, Ernst RR, Wüthrich K (1980) A two-dimensional nuclear Overhauser enhancement (2D NOE) experiment for the elucidation of complete proton-proton cross-relaxation networks in biological macromolecules. Biochem Biophys Res Commun 95:1–6CrossRefGoogle Scholar
  8. Liu M, Mao XA, Ye C, Huang H, Nicholson JK, Lindon JC (1998) Improved WATERGATE pulse sequences for solvent suppression in NMR spectroscopy. J Magn Reson 132:125–129ADSCrossRefGoogle Scholar
  9. Momot KI, Kuchel PW (2004) Convection-compensating PGSE experiment incorporating excitation-sculpting water suppression (CONVEX). J Magn Reson 169:92–101ADSCrossRefGoogle Scholar
  10. Otting G, Wüthrich K (1989) Studies of protein hydration in aqueous solution by direct NMR observation of individual protein-bound water molecules. J Am Chem Soc 111:1871–1875CrossRefGoogle Scholar
  11. Otting G, Liepinsh E, Farmer BT II, Wüthrich K (1991a) Protein hydration studied with homonuclear 3D 1H NMR experiments. J Biomol NMR 1:209–215CrossRefGoogle Scholar
  12. Otting G, Liepinsh E, Wüthrich K (1991b) Protein hydration in aqueous solution. Science 254:974–980ADSCrossRefGoogle Scholar
  13. Piotto M, Saudek V, Sklenář V (1992) Gradient-tailored excitation for single-quantum NMR spectroscopy of aqueous solutions. J Biomol NMR 2:661–665CrossRefGoogle Scholar
  14. Sklenář V, Piotto M, Leppik R, Saudek V (1993) Gradient-tailored water suppression for 1H–15N HSQC experiments optimized to retain full sensitivity. J Magn Reson 102A:241–245ADSCrossRefGoogle Scholar
  15. Stejskal EO, Schaefer J (1974a) Data routing in quadrature FT NMR. J Magn Reson 13:249–251ADSGoogle Scholar
  16. Stejskal EO, Schaefer J (1974b) Comparisons of quadrature and single-phase Fourier transform NMR. J Magn Reson 14:160–169ADSGoogle Scholar
  17. Tanner JE (1970) Use of the stimulated echo in nmr diffusion studies. J Chem Phys 52:2523–2526ADSCrossRefGoogle Scholar
  18. Wüthrich K (1986) 2D NMR with biopolymers. In: Bradbury EM, Nicolini C (ed) NMR in the life sciences, 1st edn. Plenum Press, New York, pp 11–22CrossRefGoogle Scholar
  19. Wüthrich K, Otting G, Liepinsh E (1992) Protein hydration in aqueous solution. Faraday Discuss 93:35–45ADSCrossRefGoogle Scholar
  20. Zheng G, Stait-Gardner T, Anil Kumar PG, Torres AM, Price WS (2008) PGSTE-WATERGATE: an STE-based PGSE NMR sequence with excellent solvent suppression. J Magn Reson 191:159–163ADSCrossRefGoogle Scholar
  21. Zheng G, Torres AM, Price WS (2016) WaterControl: self-diffusion based solvent signal suppression enhanced by selective inversion. Magn Reson Chem. doi: 10.1002/mrc.4420

Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  1. 1.Nanoscale Organisation and Dynamics Group, School of Science and HealthWestern Sydney UniversityPenrithAustralia

Personalised recommendations