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13C–13C NOESY spectra of a 480 kDa protein: solution NMR of ferritin

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Abstract

Molecular size has limited solution NMR analyses of proteins. We report 13C–13C NOESY experiments on a 480 kDa protein, the multi-subunit ferritin nanocage with gated pores. By exploiting 13C-resonance-specific chemical shifts and spin diffusion effects, we identified 75% of the amino acids, with intraresidue C–C connectivities between nuclei separated by 1–4 bonds. These results show the potential of 13C–13C NOESY for solution studies of molecular assemblies >100 kDa.

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Acknowledgements

The work was supported in part by MIUR (COFIN 2005), NIH grant DK20251 (ECT and MM) and the CHRCO Foundation (ECT). The authors are grateful to Dr. **aofeng Liu for advice on recombinant ferritin structure/function and expression and to Dr. Rainer Kümmerle for the competent insights on spectral acquisition.

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Authors and Affiliations

  1. Department of Chemistry, CERM, University of Florence, Via Luigi Sacconi, 6, Sesto Fiorentino, FI, 50019, Italy

    Manolis Matzapetakis, Paola Turano & Ivano Bertini

  2. CeBIC (Council for BioIron at CHORI), Children’s Hospital Oakland Research Institute, Oakland, CA, 94609, USA

    Manolis Matzapetakis & Elizabeth C. Theil

  3. Department of Nutritional Sciences and Toxicology, University of California-Berkeley, Berkeley, CA, 94720, USA

    Manolis Matzapetakis & Elizabeth C. Theil

Authors
  1. Manolis Matzapetakis
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  2. Paola Turano
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  3. Elizabeth C. Theil
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  4. Ivano Bertini
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Correspondence to Ivano Bertini.

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Matzapetakis, M., Turano, P., Theil, E.C. et al. 13C–13C NOESY spectra of a 480 kDa protein: solution NMR of ferritin. J Biomol NMR 38, 237–242 (2007). https://doi.org/10.1007/s10858-007-9163-9

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  • DOI: https://doi.org/10.1007/s10858-007-9163-9

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