Skip to main content
SpringerLink
Log in

A generic protein purification method for protein complex characterization and proteome exploration

  • In the Laboratory
  • Published:

From Nature Biotechnology

View current issue Submit your manuscript

Abstract

We have developed a generic procedure to purify proteins expressed at their natural level under native conditions using a novel tandem affinity purification (TAP) tag. The TAP tag allows the rapid purification of complexes from a relatively small number of cells without prior knowledge of the complex composition, activity, or function. Combined with mass spectrometry, the TAP strategy allows for the identification of proteins interacting with a given target protein. The TAP method has been tested in yeast but should be applicable to other cells or organisms.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1: The TAP strategy: rationale and testing.
Figure 2: Proteins purified with the TAP procedure are active.
Figure 3: TAP purification of the Mak31p identifies a new complex.

References

  1. Séraphin, B. EMBO J. 14, 2089–2098 (1995).

    Article  Google Scholar 

  2. Ford, C.F. et al. Protein Expr. Purif. 2, 95–107 (1991).

    Article  CAS  Google Scholar 

  3. Sheibani, N. Prep. Biochem. Biotechnol. 29, 77–90 (1999).

    Article  CAS  Google Scholar 

  4. Dougherty, W.G. et al. Virology 171, 356–364 (1989).

    Article  CAS  Google Scholar 

  5. Senger, B. et al. EMBO J. 17, 2196–2207 (1998).

    Article  CAS  Google Scholar 

  6. European Patent Application No. 98 115448,7.

  7. Gottschalk, A. et al. RNA 4, 374–393 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Riedel, N. et al. Proc. Natl. Acad. Sci. USA 83, 8097–8101 (1986).

    Article  CAS  Google Scholar 

  9. Shevchenko, A. et al. Proc. Natl. Acad. Sci. USA 93, 14440–14445 (1996).

    Article  CAS  Google Scholar 

  10. Wilm, M. et al. Nature 379, 466–469 (1996).

    Article  CAS  Google Scholar 

  11. Fortes P. et al. Genes & Devel. 13, in press (1999).

  12. Görlich, D. et al. Cell 87, 21–32 (1996).

    Article  Google Scholar 

  13. Wickner, R.B. Microbiol. Rev. 60, 250–265 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Caspary, F. et al. EMBO J. 18, 3463–3474 (1999).

    Article  CAS  Google Scholar 

  15. Fromont, R.M. et al. Nat. Genet. 16, 277–282 (1997).

    Article  Google Scholar 

  16. http://www.embl-heidelberg.de/ExternalInfo/seraphin/TAP.html.

  17. Puig, O. et al. Yeast 14, 1139–1146 (1998).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank F. Caspary for testing the method and suggestions on TEV cleavage, G. Stier for TEV protease, and E. Bouveret, F. Caspary, P. Lopez, O. Puig, R. Ramirez-Morales, I. Mattaj, and members of EMBL for support and/or comments on the manuscript. The German Technology Ministry (BMFF) and Glaxo-Wellcome partially supported the M.M. laboratory at EMBL.

Author information

Author notes

  1. Matthias Mann

    Present address: University of Southern Denmark, Campusvej 55, DK-5230, Odense, Denmark

Authors and Affiliations

  1. European Molecular Biology Laboratory, Meyerhofstrasse 1, Heidelberg, D-69117, Germany

    Guillaume Rigaut, Anna Shevchenko, Berthold Rutz, Matthias Wilm, Matthias Mann & Bertrand Séraphin

Authors
  1. Guillaume Rigaut
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anna Shevchenko
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Berthold Rutz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Matthias Wilm
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Matthias Mann
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bertrand Séraphin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bertrand Séraphin.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rigaut, G., Shevchenko, A., Rutz, B. et al. A generic protein purification method for protein complex characterization and proteome exploration. Nat Biotechnol 17, 1030–1032 (1999). https://doi.org/10.1038/13732

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/13732

  • Springer Nature America, Inc.

This article is cited by

Search

Navigation