Advertisement

Journal of Biomolecular NMR

, Volume 37, Issue 3, pp 195–204 | Cite as

Mixed-time parallel evolution in multiple quantum NMR experiments: sensitivity and resolution enhancement in heteronuclear NMR

  • Jinfa Ying
  • Jordan H. Chill
  • John M. Louis
  • Ad Bax
Article

Abstract

A new strategy is demonstrated that simultaneously enhances sensitivity and resolution in three- or higher-dimensional heteronuclear multiple quantum NMR experiments. The approach, referred to as mixed-time parallel evolution (MT-PARE), utilizes evolution of chemical shifts of the spins participating in the multiple quantum coherence in parallel, thereby reducing signal losses relative to sequential evolution. The signal in a given PARE dimension, t 1, is of a non-decaying constant-time nature for a duration that depends on the length of t 2, and vice versa, prior to the onset of conventional exponential decay. Line shape simulations for the 1H–15N PARE indicate that this strategy significantly enhances both sensitivity and resolution in the indirect 1H dimension, and that the unusual signal decay profile results in acceptable line shapes. Incorporation of the MT-PARE approach into a 3D HMQC-NOESY experiment for measurement of HN–HN NOEs in KcsA in SDS micelles at 50°C was found to increase the experimental sensitivity by a factor of 1.7±0.3 with a concomitant resolution increase in the indirectly detected 1H dimension. The method is also demonstrated for a situation in which homonuclear 13C–13C decoupling is required while measuring weak H3′–2′OH NOEs in an RNA oligomer.

Keywords

KcsA Mixed-time Multiple quantum NMR NOESY Parallel evolution Resolution enhancement RNA Sensitivity enhancement 

Notes

Acknowledgements

We thank Ed Nikonowicz (Rice U.) for the Helix-35 RNA sample. This work was supported by the Intramural Research Program of the NIDDK, NIH, and by the Intramural Antiviral Target Program of the Office of the Director, NIH.

Supplementary material

10858_2006_9120_MOESM1_ESM.pdf (202 kb)
(PDF 204 kb)

References

  1. Bax A, Kay LE, Sparks SW, Torchia DA (1989) Line narrowing of amide proton resonances in 2D NMR-spectra of proteins. J Am Chem Soc 111:408–409CrossRefGoogle Scholar
  2. Chill JH, Louis JM, Baber JL, Bax A (2006a) Measurement of 15N relaxation in the detergent-solubilized tetrameric KcsA potassium channel. J Biomol NMR 36:123–136CrossRefGoogle Scholar
  3. Chill JH, Louis JM, Miller C, Bax A (2006b) NMR study of tetrameric KcsA potassium channel in detergent micelles. Protein Sci 15:684–698CrossRefGoogle Scholar
  4. Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) NMRpipe – a multidimensional spectral processing system based on Unix pipes. J Biomol NMR 6:277–293CrossRefGoogle Scholar
  5. Doyle DA, Cabral JM, Pfuetzner RA, Kuo AL, Gulbis JM, Cohen SL, Chait BT, MacKinnon R (1998) The structure of the potassium channel: molecular basis of K+ conduction and selectivity. Science 280:69–77CrossRefADSGoogle Scholar
  6. Geen H, Freeman R (1991) Band-selective radiofrequency pulses. J Magn Reson 93:93–141Google Scholar
  7. Griesinger C, Sørensen OW, Ernst RR (1986) Correlation of connected transitions by two-dimensional NMR spectroscopy. J Chem Phys 85:6837–6852CrossRefADSGoogle Scholar
  8. Griffey RH, Redfield AG (1987) Proton-detected heteronuclear edited and correlated nuclear-magnetic-resonance and nuclear Overhauser effect in solution. Q Rev Biophys 19:51–82CrossRefGoogle Scholar
  9. Grzesiek S, Anglister J, Bax A (1993) Correlation of backbone amide and aliphatic side-chain resonances in 13C/15N-enriched proteins by isotropic mixing of 13C magnetization. J Magn Reson Ser B 101:114–119CrossRefGoogle Scholar
  10. Grzesiek S, Bax A (1995) Spin-locked multiple quantum coherence for signal enhancement in heteronuclear multidimensional NMR experiments. J Biomol NMR 6:335–339CrossRefGoogle Scholar
  11. Gschwind RM, Gemmecker G, Kessler H (1998) A spin system labeled and highly resolved ed-H(CCO)NH-TOCSY experiment for the facilitated assignment of proton side chains in partially deuterated samples. J Biomol NMR 11:191–198CrossRefGoogle Scholar
  12. Ikura M, Kay LE, Bax A (1990) A novel approach for sequential assignment of 1H, 13C, and 15N spectra of larger proteins: heteronuclear triple-resonance three-dimensional NMR spectroscopy. Application to calmodulin. Biochemistry 29:4659–4667CrossRefGoogle Scholar
  13. Kupče E, Freeman R (1995) Adiabatic pulses for wideband inversion and broadband decoupling. J Magn Reson A 115:273–276CrossRefGoogle Scholar
  14. Logan TM, Olejniczak ET, Xu RX, Fesik SW (1993) A general-method for assigning NMR spectra of denatured proteins using 3D HC(CO)Nh ToCSY triple resonance experiments. J Biomol NMR 3(2):225–231CrossRefGoogle Scholar
  15. Marino JP, Diener JL, Moore PB, Griesinger C (1997) Multiple-quantum coherence dramatically enhances the sensitivity of CH and CH2 correlations in uniformly 13C-labeled RNA. J Am Chem Soc 119:7361–7366Google Scholar
  16. Nikonowicz EP, Sirr A, Legault P, Jucker FM, Baer LM, Pardi A (1992) Preparation of 13C and 15N labelled RNAs for heteronuclear multi-dimensional NMR studies. Nucleic Acids Res 20(17):4507–4513CrossRefGoogle Scholar
  17. Ottiger M, Delaglio F, Bax A (1998) Measurement of J and Dipolar couplings from simplified two-dimensional NMR spectra. J Magn Reson 131:373–378CrossRefADSGoogle Scholar
  18. Pervushin K, Eletsky A (2003) A new strategy for backbone resonance assignment in large proteins using a MQ-HACACO experiment. J Biomol NMR 25:147–152CrossRefGoogle Scholar
  19. Piotto M, Saudek V, Sklenár V (1992) Gradient-tailored excitation for single-quantum NMR spectroscopy of aqueous solutions. J Biomol NMR 2(6):661–665CrossRefGoogle Scholar
  20. Plateau P, Gueron M (1982) Exchangeable proton NMR without base-line distortion, using new strong-pulse sequences. J Am Chem Soc 104:7310–7311CrossRefGoogle Scholar
  21. Shang Z, Swapna GVT, Rios CB, Montelione GT (1997) Sensitivity enhancement of triple-resonance protein NMR spectra by proton evolution of multiple-quantum coherences using a simultaneous 1H and 13C constant-time evolution period. J Am Chem Soc 119:9274–9278CrossRefGoogle Scholar
  22. Ying J, Bax A (2006) Determination of 2′-OH proton positions in helical RNA by simultaneously measured heteronuclear scalar couplings and NOEs. J Am Chem Soc 128:8372–8373CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Jinfa Ying
    • 1
  • Jordan H. Chill
    • 1
  • John M. Louis
    • 1
  • Ad Bax
    • 1
  1. 1.Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney DiseasesNational Institutes of HealthBethesdaUSA

Personalised recommendations