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

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Abstract

We introduce an efficient approach for sequential protein backbone assignment based on two complementary proton-detected 4D solid-state NMR experiments that correlate \( {\text{H}}_{{\text{i}}}^{{\text{N}}} \)/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.

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Acknowledgments

We thank Karin Giller and Brigitta Angerstein for expert technical assistance. This work was supported by the Max Planck Society, the Leibniz-Institut für Molekulare Pharmakologie and the DFG (Emmy Noether Fellowship to A.L). A.L. and S.X. acknowledge funding from the CRC803 (DFG).

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Authors and Affiliations

  1. Department of NMR-based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077, Göttingen, Germany

    ShengQi Xiang, Veniamin Chevelkov, Stefan Becker & Adam Lange

  2. Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Str. 10, 13125, Berlin, Germany

    Veniamin Chevelkov & Adam Lange

  3. Institut für Biologie, Humboldt-Universität zu Berlin, Berlin, Germany

    Adam Lange

Authors
  1. ShengQi Xiang
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  2. Veniamin Chevelkov
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  3. Stefan Becker
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  4. Adam Lange
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Correspondence to Adam Lange.

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Xiang, S., Chevelkov, V., Becker, S. et al. Towards automatic protein backbone assignment using proton-detected 4D solid-state NMR data. J Biomol NMR 60, 85–90 (2014). https://doi.org/10.1007/s10858-014-9859-6

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  • DOI: https://doi.org/10.1007/s10858-014-9859-6

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