Solid-state NMR spectroscopy of 10% 13C labeled ubiquitin: spectral simplification and stereospecific assignment of isopropyl groups

Abstract

We describe the simplification of 13C–13C correlation spectra obtained from a microcrystalline protein sample expressed on a growth medium of 10% fully 13C labeled glucose diluted in 90% natural abundance glucose as compared to a fully labeled sample. Such a labeling scheme facilitates the backbone and side-chain resonance assignment of Phe, Tyr, His, Asp, Asn, Ile, Lys and Pro and yields an unambiguous stereospecific assignment of the valine Cγ1, Cγ2 13C resonances and of Leucine Cδ2.

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Acknowledgements

We acknowledge scientific discussions with Matthias Ernst, Ansgar Siemer, Stephan Grzesiek, Hans-Jürgen Sass, David Sargent and Alvar Gossert. We thank Andreas Hunkeler and Urban Meier for technical assistance and Barth van Rossum for carefully reading the manuscript. The research was supported by the ETH Zurich (TH grant 30/03-1) and the Swiss National Science Foundation (SNF).

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Correspondence to Beat H. Meier.

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Schubert, M., Manolikas, T., Rogowski, M. et al. Solid-state NMR spectroscopy of 10% 13C labeled ubiquitin: spectral simplification and stereospecific assignment of isopropyl groups. J Biomol NMR 35, 167–173 (2006). https://doi.org/10.1007/s10858-006-9025-x

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Key words

  • MAS
  • resonance assignment
  • solid state NMR
  • stereoselective isotope labeling
  • stereospecific assignment