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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References
Séraphin, B. EMBO J. 14, 2089–2098 (1995).
Ford, C.F. et al. Protein Expr. Purif. 2, 95–107 (1991).
Sheibani, N. Prep. Biochem. Biotechnol. 29, 77–90 (1999).
Dougherty, W.G. et al. Virology 171, 356–364 (1989).
Senger, B. et al. EMBO J. 17, 2196–2207 (1998).
European Patent Application No. 98 115448,7.
Gottschalk, A. et al. RNA 4, 374–393 (1998).
Riedel, N. et al. Proc. Natl. Acad. Sci. USA 83, 8097–8101 (1986).
Shevchenko, A. et al. Proc. Natl. Acad. Sci. USA 93, 14440–14445 (1996).
Wilm, M. et al. Nature 379, 466–469 (1996).
Fortes P. et al. Genes & Devel. 13, in press (1999).
Görlich, D. et al. Cell 87, 21–32 (1996).
Wickner, R.B. Microbiol. Rev. 60, 250–265 (1996).
Caspary, F. et al. EMBO J. 18, 3463–3474 (1999).
Fromont, R.M. et al. Nat. Genet. 16, 277–282 (1997).
http://www.embl-heidelberg.de/ExternalInfo/seraphin/TAP.html.
Puig, O. et al. Yeast 14, 1139–1146 (1998).
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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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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DOI: https://doi.org/10.1038/13732
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