Abstract
High signal to noise is a necessity for the quantification of NMR spectral parameters to be translated into accurate and precise restraints on protein structure and dynamics. An important source of long-range structural information is obtained from 1H–1H residual dipolar couplings (RDCs) measured for weakly aligned molecules. For sensitivity reasons, such measurements are generally performed on highly deuterated protein samples. Here we show that high sensitivity is also obtained for protonated protein samples if the pulse schemes are optimized in terms of longitudinal relaxation efficiency and J-mismatch compensated coherence transfer. The new sensitivity-optimized quantitative J-correlation experiment yields important signal gains reaching factors of 1.5 to 8 for individual correlation peaks when compared to previously proposed pulse schemes.
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Acknowledgments
This work was supported by the Commissariat à l’Energie Atomique, the Centre National de la Recherche Scientifique, the French Research Agency (ANR), Human Frontier Science Program Organization and the European Commission (EU-NMR). P.S. and R.S. acknowledge support from the French ministry of education, research, and technology. We thank Beate Bersch, Isabel Ayala, and Jacques Covès (IBS Grenoble) for the preparation of the isotope-labeled protein samples.
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Paul Schanda and Ewen Lescop contributed equally to this work.
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Schanda, P., Lescop, E., Falge, M. et al. Sensitivity-optimized experiment for the measurement of residual dipolar couplings between amide protons. J Biomol NMR 38, 47–55 (2007). https://doi.org/10.1007/s10858-006-9138-2
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DOI: https://doi.org/10.1007/s10858-006-9138-2