Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Brief Communication
  • Published:

SUnSET, a nonradioactive method to monitor protein synthesis

Abstract

We developed a nonradioactive fluorescence-activated cell sorting–based assay, called surface sensing of translation (SUnSET), which allows the monitoring and quantification of global protein synthesis in individual mammalian cells and in heterogeneous cell populations. We demonstrate here, using mouse dendritic and T cells as a model, that SUnSET offers a technical alternative to classical radioactive labeling methods for the study of mRNA translation and cellular activation.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Translation monitoring using puromycin-labeled proteins.
Figure 2: Translation quantification in different cell populations using SUnSET.

Similar content being viewed by others

References

  1. Sonenberg, N. & Hinnebusch, A.G. Mol. Cell 28, 721–729 (2007).

    Article  CAS  Google Scholar 

  2. Sabatini, D.M. Nat. Rev. Cancer 6, 729–773 (2006).

    Article  CAS  Google Scholar 

  3. Nathans, D. Fed. Proc. 23, 984–989 (1964).

    CAS  PubMed  Google Scholar 

  4. Hansen, W.J., Lingappa, V.R. & Welch, W.J. J. Biol. Chem. 269, 26610–26613 (1994).

    CAS  PubMed  Google Scholar 

  5. Nemoto, N., Miyamoto-Sato, E. & Yanagawa, H. FEBS Lett. 462, 43–46 (1999).

    Article  CAS  Google Scholar 

  6. Starck, S.R. et al. Chem. Biol. 11, 999–1008 (2004).

    Article  CAS  Google Scholar 

  7. Lelouard, H. et al. J. Cell Biol. 164, 667–675 (2004).

    Article  CAS  Google Scholar 

  8. Anderson, P. & Kedersha, N. J. Cell Biol. 172, 803–808 (2006).

    Article  CAS  Google Scholar 

  9. Hurtley, S.M. & Helenius, A. Annu. Rev. Cell Biol. 5, 277–307 (1989).

    Article  CAS  Google Scholar 

  10. Lelouard, H. et al. J. Cell Biol. 179, 1427–1439 (2007).

    Article  CAS  Google Scholar 

  11. Zheng, Y. et al. J. Immunol. 178, 2163–2170 (2007).

    Article  CAS  Google Scholar 

  12. Choo, A.Y. et al. Proc. Natl. Acad. Sci. USA 105, 17414–17419 (2008).

    Article  CAS  Google Scholar 

  13. Smith, W.B. et al. Neuron 45, 765–779 (2005).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

E.K.S. and G.C. are supported by fellowships from the Ministère de la Recherche and la Fondation pour la Recherche Médicale. This work is supported by grants to P.P. from Ligue National Contre le Cancer, the Human Frontier Science Program and the European Network of Excellence DC-THERA. We thank E. Gatti for useful discussions and manuscript proofreading and members of the Plateforme d'Imagerie Commune du Site de Luminy (PICsL) for expert technical assistance.

Author information

Authors and Affiliations

Authors

Contributions

E.K.S., G.C. and M.C. performed experiments. E.K.S. and P.P. designed experiments and wrote the paper.

Corresponding author

Correspondence to Philippe Pierre.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–5, Supplementary Methods (PDF 2887 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schmidt, E., Clavarino, G., Ceppi, M. et al. SUnSET, a nonradioactive method to monitor protein synthesis. Nat Methods 6, 275–277 (2009). https://doi.org/10.1038/nmeth.1314

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth.1314

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing