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

Nature Biotechnology volume 17, pages 1030–1032 (1999)Cite this article

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.

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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.

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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.

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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

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  1. Guillaume Rigaut
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  2. Anna Shevchenko
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  3. Berthold Rutz
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  4. Matthias Wilm
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  6. Bertrand Séraphin
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Correspondence to Bertrand Séraphin.

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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

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