Abstract
Resonance assignment is the first step in NMR structure determination. For magic angle spinning NMR, this is typically achieved with a set of heteronuclear correlation experiments (NCaCX, NCOCX, CONCa) that utilize SPECIFIC-CP 15N–13C transfers. However, the SPECIFIC-CP transfer efficiency is often compromised by molecular dynamics and probe performance. Here we show that one-bond ZF-TEDOR 15N–13C transfers provide simultaneous NCO and NCa correlations with at least as much sensitivity as SPECIFIC-CP for some non-crystalline samples. Furthermore, a 3D ZF-TEDOR-CC experiment provides heteronuclear sidechain correlations and robustness with respect to proton decoupling and radiofrequency power instabilities. We demonstrate transfer efficiencies and connectivities by application of 3D ZF-TEDOR-DARR to a model microcrystalline protein, GB1, and a less ideal system, GvpA in intact gas vesicles.
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Acknowledgments
We thank Dr. Marvin Bayro, Dr. Galia Debelouchina, Dr. Vladimir Michaelis, Dr. Christopher Turner and Dr. David Ruben for insightful discussions, Mr. Ajay Thakkar, Mr. Mike Mullins, and Dr. David Ruben for technical assistance, Lindsay Clark for preparing the GB1 sample, and Marina Belenky for preparing the gas vesicle sample. Research reported in this publication was supported by the National Institute of Biomedical Imaging and Bioengineering of the National Institutes of Health under awards EB001035, EB-001960 and EB-002926.
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Daviso, E., Eddy, M.T., Andreas, L.B. et al. Efficient resonance assignment of proteins in MAS NMR by simultaneous intra- and inter-residue 3D correlation spectroscopy. J Biomol NMR 55, 257–265 (2013). https://doi.org/10.1007/s10858-013-9707-0
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DOI: https://doi.org/10.1007/s10858-013-9707-0