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Towards automatic protein backbone assignment using proton-detected 4D solid-state NMR data.

MPG-Autoren
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Chevelkov,  V.
Department of NMR Based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Becker,  S.
Department of NMR Based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Lange,  A.
Department of NMR Based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

Volltexte (frei zugänglich)

2069432.pdf
(Verlagsversion), 984KB

Ergänzendes Material (frei zugänglich)

2069432_Suppl.pdf
(Ergänzendes Material), 616KB

Zitation

Xiang, S. Q., Chevelkov, V., Becker, S., & Lange, A. (2014). Towards automatic protein backbone assignment using proton-detected 4D solid-state NMR data. Journal of Biomolecular NMR, 60(2-3), 85-90. doi:10.1007/s10858-014-9859-6.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-0024-246E-D
Zusammenfassung
We introduce an efficient approach for sequential protein backbone assignment based on two complementary proton-detected 4D solid-state NMR experiments that correlate HNi /Ni with CAi/COi or CAi−1/COi−1. The resulting 4D spectra exhibit excellent sensitivity and resolution and are amenable to (semi-)automatic assignment approaches. This strategy allows to obtain sequential connections with high confidence as problems related to peak overlap and multiple assignment possibilities are avoided. Non-uniform sampling schemes were implemented to allow for the acquisition of 4D spectra within a few days. Rather moderate hardware requirements enable the successful demonstration of the method on deuterated type III secretion needles using a 600 MHz spectrometer at a spinning rate of 25 kHz.