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Atomic-resolution monitoring of protein maturation in live human cells by NMR

Nature Chemical Biology volume 9, pages 297–299 (2013)Cite this article

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Abstract

We use NMR directly in live human cells to describe the complete post-translational maturation process of human superoxide dismutase 1 (SOD1). We follow, at atomic resolution, zinc binding, homodimer formation and copper uptake, and discover that copper chaperone for SOD1 oxidizes the SOD1 intrasubunit disulfide bond through both copper-dependent and copper-independent mechanisms. Our approach represents a new strategy for structural investigation of endogenously expressed proteins in a physiological (cellular) environment.

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Figure 1: Zn(II) added to the culture medium promotes binding of one Zn2+ ion per apo-SOD1 subunit in the cytoplasm.
Figure 2: Cu(II) addition to the culture medium induces Cu(I) binding to a fraction of cytoplasmic SOD1.
Figure 3: The redox state of SOD1 is influenced by both copper binding and the presence of CCS.

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References

  1. Reckel, S., Hänsel, R., Löhr, F. & Dötsch, V. Prog. Nucl. Magn. Reson. Spectrosc. 51, 91–101 (2007).

    Article  CAS  Google Scholar 

  2. Burz, D.S. & Shekhtman, A. PLoS ONE 3, e2571 (2008).

    Article  Google Scholar 

  3. Inomata, K. et al. Nature 458, 106–109 (2009).

    Article  CAS  Google Scholar 

  4. Ogino, S. et al. J. Am. Chem. Soc. 131, 10834–10835 (2009).

    Article  CAS  Google Scholar 

  5. Selenko, P. et al. Nat. Struct. Mol. Biol. 15, 321–329 (2008).

    Article  CAS  Google Scholar 

  6. Culotta, V.C., Yang, M. & O'Halloran, T.V. Biochim. Biophys. Acta 1763, 747–758 (2006).

    Article  CAS  Google Scholar 

  7. Michiels, C., Raes, M., Toussaint, O. & Remacle, J. Free Radic. Biol. Med. 17, 235–248 (1994).

    Article  CAS  Google Scholar 

  8. Lindberg, M.J., Tibell, L. & Oliveberg, M. Proc. Natl. Acad. Sci. USA 99, 16607–16612 (2002).

    Article  CAS  Google Scholar 

  9. Banci, L. et al. Proc. Natl. Acad. Sci. USA 104, 11263–11267 (2007).

    Article  CAS  Google Scholar 

  10. Aricescu, A.R., Lu, W. & Jones, E.Y. Acta Crystallogr. D Biol. Crystallogr. 62, 1243–1250 (2006).

    Article  Google Scholar 

  11. Chang, L.Y., Slot, J.W., Geuza, H.J. & Crapo, J.D. J. Cell Biol. 107, 2169–2179 (1988).

    Article  CAS  Google Scholar 

  12. Sturtz, L.A., Diekert, K., Jensen, L.T., Lill, R. & Culotta, V.C. J. Biol. Chem. 276, 38084–38089 (2001).

    CAS  PubMed  Google Scholar 

  13. Banci, L., Bertini, I., Cramaro, F., Del Conte, R. & Viezzoli, M.S. Biochemistry 42, 9543–9553 (2003).

    Article  CAS  Google Scholar 

  14. Banci, L., Bertini, I., Cantini, F., D'Amelio, N. & Gaggelli, E. J. Biol. Chem. 281, 2333–2337 (2006).

    Article  CAS  Google Scholar 

  15. Banci, L., Barbieri, L., Bertini, I., Cantini, F. & Luchinat, E. PLoS ONE 6, e23561 (2011).

    Article  CAS  Google Scholar 

  16. Kim, B.E., Nevitt, T. & Thiele, D.J. Nat. Chem. Biol. 4, 176–185 (2008).

    Article  CAS  Google Scholar 

  17. Puig, S. & Thiele, D.J. Curr. Opin. Chem. Biol. 6, 171–180 (2002).

    Article  CAS  Google Scholar 

  18. Banci, L. et al. Nature 465, 645–648 (2010).

    Article  CAS  Google Scholar 

  19. Schmidt, P.J. et al. J. Biol. Chem. 274, 23719–23725 (1999).

    Article  CAS  Google Scholar 

  20. Furukawa, Y., Torres, A.S. & O'Halloran, T.V. EMBO J. 23, 2872–2881 (2004).

    Article  CAS  Google Scholar 

  21. Carroll, M.C. et al. Proc. Natl. Acad. Sci. USA 101, 5964–5969 (2004).

    Article  CAS  Google Scholar 

  22. Leitch, J.M., Yick, P.J. & Culotta, V.C. J. Biol. Chem. 284, 24679–24683 (2009).

    Article  CAS  Google Scholar 

  23. Proescher, J.B., Son, M., Elliott, J.L. & Culotta, V.C. Hum. Mol. Genet. 17, 1728–1737 (2008).

    Article  CAS  Google Scholar 

  24. Banci, L. et al. Proc. Natl. Acad. Sci. USA 109, 13555–13560 (2012).

    Article  CAS  Google Scholar 

  25. Reckel, S., Lopez, J.J., Löhr, F., Glaubitz, C. & Dötsch, V. ChemBioChem 13, 534–537 (2012).

    Article  CAS  Google Scholar 

  26. Schanda, P. & Brutscher, B. J. Am. Chem. Soc. 127, 8014–8015 (2005).

    Article  CAS  Google Scholar 

  27. Freshney, R. Culture of Animal Cells: A Manual of Basic Technique, 2nd edn. (Wiley-Liss, 1987).

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Acknowledgements

We would like to thank E.Y. Jones and D.I. Stuart for critically reading the manuscript and providing advice. This work was supported by the Access to Research Infrastructures activities in the Seventh Framework Programme of the European Commission (Bio-NMR contract 261863 and P-CUBE contract 227764), by the Italian grant program MIUR-PRIN 2009 'Biologia strutturale meccanicistica: avanzamenti metodologici e biologici' and by the European Integrated Structural Biology Infrastructure (INSTRUCT), which is part of the European Strategy Forum on Research Infrastructures (ESFRI) and supported by national member subscriptions. Specifically, we thank the INSTRUCT Core Centres CERM (Italy) and the University of Oxford. A.R.A. was supported by a UK Medical Research Council Career Development Award fellowship.

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Author notes

  1. Ivano Bertini: Deceased.

Authors and Affiliations

  1. CERM, Magnetic Resonance Center, University of Florence, Florence, Italy

    Lucia Banci, Letizia Barbieri, Ivano Bertini, Enrico Luchinat & Erica Secci

  2. Department of Chemistry, University of Florence, Florence, Italy

    Lucia Banci & Ivano Bertini

  3. Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK

    Yuguang Zhao & A Radu Aricescu

Authors
  1. Lucia Banci
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  2. Letizia Barbieri
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  3. Ivano Bertini
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  4. Enrico Luchinat
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  5. Erica Secci
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  6. Yuguang Zhao
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  7. A Radu Aricescu
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Contributions

L. Banci, I.B. and A.R.A. conceived the work; L. Banci, L. Barbieri, E.L. and A.R.A. designed the experiments; L. Barbieri and Y.Z. grew and transfected the human cells; L. Barbieri cloned the genes and produced the cell samples of SOD1, CCS, Atox1, Grx1 and Trx; E.S. produced the cell sample of Mia40; L. Barbieri and E.L. produced and analyzed the E. coli cell samples; E.L. performed the NMR experiments and analyzed the data; L. Banci, L. Barbieri, I.B., E.L. and A.R.A. wrote the paper.

Corresponding authors

Correspondence to Lucia Banci or A Radu Aricescu.

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The authors declare no competing financial interests.

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Banci, L., Barbieri, L., Bertini, I. et al. Atomic-resolution monitoring of protein maturation in live human cells by NMR. Nat Chem Biol 9, 297–299 (2013). https://doi.org/10.1038/nchembio.1202

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