一种用于蛋白质复合物表征和蛋白质组探索的通用蛋白质纯化方法。

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

作者信息

Rigaut G, Shevchenko A, Rutz B, Wilm M, Mann M, Séraphin B

机构信息

European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.

出版信息

Nat Biotechnol. 1999 Oct;17(10):1030-2. doi: 10.1038/13732.

DOI:10.1038/13732

PMID:10504710

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.

摘要

我们开发了一种通用程序，可利用新型串联亲和纯化（TAP）标签在天然条件下纯化以天然水平表达的蛋白质。TAP标签能够从相对少量的细胞中快速纯化复合物，而无需事先了解复合物的组成、活性或功能。结合质谱分析，TAP策略可用于鉴定与给定目标蛋白相互作用的蛋白质。TAP方法已在酵母中进行了测试，但应该适用于其他细胞或生物体。