Recent advances in stable-isotope enrichment and heteronuclear multidimensional NMR techniques have transformed NMR into a more powerful and versatile method for the structural and dynamic characterization of biomolecules. Current efforts still focus on improving the methodology to increase the number of systems amenable to NMR analysis, to generate better structures and to investigate biomolecular motions in solution.
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Highly efficient heteronuclear polarization transfer using dipolar-echo edited R-symmetry sequences in solid-state NMR.
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State Key Laboratory of Catalysis, Dalian National Laboratory for Clean Energy, Dalian Institute of Chemical Physics, Chinese Academy of Sciences Zhongshan Road 457 Dalian 116023 China
In solid-state NMR, dipolar-based heteronuclear polarization transfer has been extensively used for sensitivity enhancement and multidimensional correlations, but its efficiency often suffers from undesired spin interactions and hardware limitations. Herein, we propose a novel dipolar-echo edited R-symmetry (DEER) sequence, which is further incorporated into the INEPT-type scheme, dubbed DEER-INEPT, for achieving highly efficient heteronuclear polarization transfer. Numerical simulations and NMR experiments demonstrate that DEER-INEPT offers significantly improved robustness, enabling efficient polarization transfer under a wide range of MAS conditions, from slow to ultrafast rates, outperforming existing methods.
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Department of Chemistry and Francis Bitter Magnet Laboratory, Massachusetts Institute of Technology, Cambridge, MA 02139, United States. Electronic address:
In magic angle spinning (MAS) experiments involving uniformly C/N labeled proteins, C-C and C-N dipolar recoupling experiments are now routinely used to measure direct dipole-dipole couplings that constrain distances and torsion angles and determine molecular structures. When the distances are short (<4 Å), the direct couplings dominate the evolution of the spin system, and the C-C and C-N J-couplings (scalar couplings) are ignored. However, for structurally interesting >4 Å distances, the dipolar and J-couplings are generally of comparable magnitude, and the variation in J must be included in order to optimize the precision of the experiment.
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Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0520, U.S.A. Electronic address:
Dynamic nuclear polarization (DNP) enhanced magic angle spinning (MAS) solid-state NMR carried out at 25 K enables rapid acquisition of multi-dimensional C-N correlation spectra for protein structure studies and resonance assignment. Under commonly used DNP conditions, solvent deuteration reduces H-N cross polarization (CP) efficiencies, necessitates more careful optimization, and requires longer high-power N radio-frequency pulses. The sensitivity of 2D heteronuclear correlation experiments is potentially impaired.
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