Daulat Avais M, Maurice Pascal, Jockers Ralf
Institut Cochin, Université Paris Descartes, INSERM, Paris, France.
Methods Mol Biol. 2011;746:399-409. doi: 10.1007/978-1-61779-126-0_23.
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.
首个串联亲和纯化（TAP）方案于1999年被描述。它最初设计用于酵母RNA剪接中蛋白质复合物的纯化，其应用迅速扩展至酵母和哺乳动物细胞的全蛋白质组分析。最近，TAP已应用于G蛋白偶联受体（GPCR）相关蛋白复合物（GAPC）的纯化。这种方法对GPCR特别有吸引力，因为天然的七跨膜结构被用作诱饵，从在生理水平表达受体的哺乳动物细胞中纯化GAPC。本文介绍了应用于GPCR的TAP方法的详细方案。
