Abstract
Experimental sensitivity remains a major drawback for the application of NMR spectroscopy to fragile and low concentrated biomolecular samples. Here we describe an efficient polarization enhancement mechanism in longitudinal-relaxation enhanced fast-pulsing triple-resonance experiments. By recovering undetectable 1H polarization originating from longitudinal relaxation during the pulse sequence, the steady-state 15N polarization becomes enhanced by up to a factor of ~5 with respect to thermal equilibrium yielding significant sensitivity improvements compared to conventional schemes. The benefits of BEST-TROSY experiments at high magnetic field strength are illustrated for various protein applications, but they will be equally useful for other protonated macromolecular systems.
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Acknowledgments
We thank Drs J. Boisbouvier, S. Hediger, and M. Plevin for stimulating discussion and critical reading of this manuscript, and I. Ayala for expert protein production. This work was supported by the Commissariat à l’Energie Atomique, the Centre National de la Recherche Scientifique, the University of Grenoble, and the French research agency (grant ANR 08-BLAN-210) and the European Commission (FP7-I3 BIO-NMR contract No. 261863).
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Favier, A., Brutscher, B. Recovering lost magnetization: polarization enhancement in biomolecular NMR. J Biomol NMR 49, 9–15 (2011). https://doi.org/10.1007/s10858-010-9461-5
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DOI: https://doi.org/10.1007/s10858-010-9461-5