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Use of biosynthetic fractional 13C-labeling for backbone NMR assignment of proteins

Journal of Biomolecular NMR volume 37, pages 187–193 (2007)Cite this article

Abstract

We describe a simple approach to classify amino acid residue types in NMR spectra of proteins for supporting the backbone resonance assignments. It makes use of the differences in biosynthetic pathways of the 20 amino acids in Escherichia coli. Therefore, it is distinct from the parameters routinely exploited in the backbone resonance assignment such as chemical shifts and spin topology information. The combination of biosynthetically directed fractional 13C-labeling and uniform 15N-labeling enables us to obtain both residue-type specific information and sequential connectivities from a single protein sample. The residue-type classification exploiting biosynthetic pathways can be used for accelerating the conventional backbone assignment procedure.

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Acknowledgments

The authors thank Sara Züger, Dalibor Hrstka, and Katja Karjalainen for the preparation of the samples. This work is supported by the Academy of Finland and by the Biocentrum Helsinki. JF gratefully acknowledges support from the Human Science Frontier Program.

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Affiliations

  1. Research Program in Structural Biology and Biophysics, Institute of Biotechnology, University of Helsinki, P.O. Box 65, Helsinki, 00014, Finland

    Hideo Iwai

  2. Zentrum für Molekulare Biologie, Universität Heidelberg, Im Neuenheimer Feld 282, Heidelberg, 69120, Germany

    Jocelyne Fiaux

Authors
  1. Hideo Iwai
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  2. Jocelyne Fiaux
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Correspondence to Hideo Iwai.

Electronic supplementary material

A table ofsIvalues measured for the samples of 434(1–63) produced under different culture conditions

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Iwai, H., Fiaux, J. Use of biosynthetic fractional 13C-labeling for backbone NMR assignment of proteins. J Biomol NMR 37, 187–193 (2007). https://doi.org/10.1007/s10858-006-9124-8

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  • DOI: https://doi.org/10.1007/s10858-006-9124-8

Keywords

  • Isotope labeling
  • Backbone resonance assignment
  • Amino acid metabolism