Resolution enhancement by homonuclear J-decoupling: application to three-dimensional solid-state magic angle spinning NMR spectroscopy |
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Authors: | Lichi Shi Xiaohu Peng Mumdooh A M Ahmed Dale Edwards Leonid S Brown Vladimir Ladizhansky |
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Institution: | Department of Physics and Biophysics Interdepartmental Group, University of Guelph, 50 Stone Road East, Guelph, ON, Canada N1G 2W1. |
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Abstract: | We describe a simple protocol to achieve homonuclear J-decoupling in the indirect dimensions of multidimensional experiments, and to enhance spectral resolution of the backbone Calpha carbons in the 3D NCACX experiment. In the proposed protocol, the refocusing of the Calpha-CO homonuclear J-couplings is achieved by applying an off-resonance selective pi pulse to the CO spectral region in the middle of Calpha chemical shift evolution. As is commonly used in solution NMR, a compensatory echo period is used to refocus the unwanted chemical shift evolution of Calpha spins, which takes place during the off-resonance selective pulse. The experiments were carried out on the beta1 immunoglobulin binding domain of protein G (GB1). In GB1, such implementation results in significantly reduced line widths, and leads to an overall sensitivity enhancement. |
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Keywords: | Solid-state NMR Magic angle spinning Uniformly 13C 15N labeled proteins Assignments Homonuclear decoupling Resolution enhancement |
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