Abstract
We present an approach for the assignment of protein NMR resonances that combines established and new concepts: (a) Based on published reduced dimensionality methods, two 5-dimensional experiments are proposed. (b) Multi-way decomposition (PRODECOMP) applied simultaneously to all acquired NMR spectra provides the assignment of resonance frequencies under conditions of very low signal-to-noise. (c) Each resulting component characterizes all spin ½ nuclei in a (doubly-labeled) CβHn–CαH–C′–NH–CαH–CβHn fragment in an unambiguous manner, such that sequentially neighboring components have about four atoms in common. (d) A new routine (SHABBA) determines correlations for all component pairs based on the common nuclei; high correlation values yield sequential chains of a dozen or more components. (e) The potentially error-prone peak picking is delayed to the last step, where it helps to place the component chains within the protein sequence, and thus to achieve the final backbone assignment. The approach was validated by achieving complete backbone resonance assignments for ubiquitin.
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Acknowledgments
This work was supported by research grants from the Swedish Research Council (621-2003-4048) and the EU (LSHG-CT-2005-018988). Data from the BioMagResBank were used.
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Staykova, D.K., Fredriksson, J., Bermel, W. et al. Assignment of protein NMR spectra based on projections, multi-way decomposition and a fast correlation approach. J Biomol NMR 42, 87–97 (2008). https://doi.org/10.1007/s10858-008-9265-z
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DOI: https://doi.org/10.1007/s10858-008-9265-z