Gradient and sensitivity enhancement of 2D TROSY with water flip-back, 3D NOESY-TROSY and TOCSY-TROSY experiments

AbstractPreviously we demonstrated a sensitivity enhancement of the original TROSY experiment by a factor of $$\sqrt 2$$ by the use of the sensitivity enhanced TROSY (en-TROSY) scheme. Here, we develop a gradient and sensitivity enhanced TROSY experiment (gs-TROSY), which is designed to select magnetization transfer pathways that suppress spectral artifacts and reduce the number of required phase cycles while having minimal loss of sensitivity. Both of these experimental methods (en-TROSY and gs- TROSY) have been combined with a water flip-back scheme which provides a further increase in sensitivity for labile NH groups by avoiding water saturation. We also apply these TROSY schemes to 3D NOESY-TROSY and 3D TOCSY-TROSY experiments.

[1]  D. States,et al.  A two-dimensional nuclear overhauser experiment with pure absorption phase in four quadrants☆ , 1982 .

[2]  F. Richards,et al.  NMR sequential assignment of Escherichia coli thioredoxin utilizing random fractional deuteriation. , 1988, Biochemistry.

[3]  A. Bax,et al.  Delineation of .alpha.-helical domains in deuteriated Staphylococcal nuclease by 2D NOE NMR spectroscopy , 1988 .

[4]  E. Geiduschek,et al.  Improving two-dimensional proton NMR NOESY spectra of a large protein by selective deuteriation , 1991 .

[5]  A. Bax,et al.  Improved linear prediction of damped NMR signals using modified forward-backward linear prediction , 1992 .

[6]  Paul A. Keifer,et al.  Pure absorption gradient enhanced heteronuclear single quantum correlation spectroscopy with improved sensitivity , 1992 .

[7]  S. Grzesiek,et al.  The Importance of Not Saturating H2o in Protein NMR : application to Sensitivity Enhancement and Noe Measurements , 1993 .

[8]  P. Domaille,et al.  An Approach to the Structure Determination of Larger Proteins Using Triple Resonance NMR Experiments in Conjunction with Random Fractional Deuteration , 1996 .

[9]  G. Zhu,et al.  Sensitivity Enhancement in Transverse Relaxation Optimized NMR Spectroscopy. , 1998, Angewandte Chemie.

[10]  G. Zhu,et al.  Gradient- and sensitivity-enhanced heteronuclear multiple-quantum correlation spectroscopy. , 1998, Journal of magnetic resonance.

[11]  H. Senn,et al.  HMQC and HSQC experiments with water flip-back optimized for large proteins , 1998, Journal of biomolecular NMR.