Protocol

Receptor Signal Transduction Protocols

Volume 746 of the series Methods in Molecular Biology pp 399-409

Date:

Tandem Affinity Purification and Identification of GPCR-Associated Protein Complexes

  • Avais M. Daulat
  • , Pascal Maurice
  • , Ralf JockersAffiliated withInstitut Cochin, Université Paris Descartes, INSERM Email author 

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Abstract

The first tandem affinity purification (TAP) protocol was described in 1999. Originally designed for the purification of protein complexes in yeast RNA splicing, its application rapidly expanded towards whole proteome analysis in yeast and mammalian cells. More recently, TAP has been applied to the purification of G protein-coupled receptor (GPCR)-associated protein complexes (GAPCs). This approach is particularly attractive for GPCRs, as the native, seven transmembrane structure is used as bait to purify GAPCs from mammalian cells expressing receptors at physiological levels. Here, a detailed protocol of the TAP method applied to GPCRs is presented.

Key words

G protein-coupled receptor Tandem affinity purification Protein complexes Proteomics