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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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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