Efficient resonance assignment of proteins in MAS NMR by simultaneous intra- and inter-residue 3D correlation spectroscopy
Author(s)Daviso, Eugenio; Eddy, Matthew Thomas; Andreas, Loren; Herzfeld, Judith; Griffin, Robert Guy
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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 [superscript 15]N–[superscript 13]C transfers. However, the SPECIFIC-CP transfer efficiency is often compromised by molecular dynamics and probe performance. Here we show that one-bond ZF-TEDOR [superscript 15]N– [superscript 13]C 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.
DepartmentMassachusetts Institute of Technology. Department of Chemistry; Francis Bitter Magnet Laboratory (Massachusetts Institute of Technology)
Journal of Biomolecular NMR
Daviso, Eugenio, Matthew T. Eddy, Loren B. Andreas, Robert G. Griffin, and Judith Herzfeld. “Efficient Resonance Assignment of Proteins in MAS NMR by Simultaneous Intra- and Inter-Residue 3D Correlation Spectroscopy.” Journal of Biomolecular NMR 55, no. 3 (January 19, 2013): 257–265.
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