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

@article{Rigaut1999AGP,
  title={A generic protein purification method for protein complex characterization and proteome exploration},
  author={Guillaume Rigaut and Anna Shevchenko and Berthold Rutz and Matthias Wilm and Matthias Mann and Bertrand S{\'e}raphin},
  journal={Nature Biotechnology},
  year={1999},
  volume={17},
  pages={1030-1032}
}
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… Expand
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The TAP method is developed as a tool that allows rapid purification under native conditions of complexes, even when expressed at their natural level, and is a very useful procedure for protein purification and proteome exploration. Expand
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A tag based on protein G and the streptavidin-binding peptide resulted in a tenfold increase in protein-complex yield and improved the specificity of the procedure, allowing purification of protein complexes that were hitherto not amenable to TAP and use of less starting material, leading to higher success rates and enabling systematic interaction proteomics projects. Expand
Identification of protein complexes from filamentous fungi with tandem affinity purification.
TLDR
This work codon-optimized tags and established TAP, previously not applicable to filamentous fungi, for the model organism Aspergillus nidulans, and identified by this method the trimeric Velvet complex VelB/VeA/LaeA or the eight subunit COP9 signalosome. Expand
Targeted purification of SnAvi-tagged proteins.
TLDR
The application of the SnAvi-tag-a TAP-tag useful in different expression systems is described, which is multifunctional and facilitates among others the in vivo visualization of tagged proteins and their cell type specific activation. Expand
Tandem affinity purification and identification of GPCR-associated protein complexes.
TLDR
A detailed protocol of the TAP method applied to GPCRs is presented and it is shown that the native, seven transmembrane structure is used as bait to purify GAPCs from mammalian cells expressing receptors at physiological levels. Expand
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