Identification of Signaling Protein Complexes by Parallel Affinity Precipitation Coupled with Mass Spectrometry

平行亲和沉淀结合质谱法鉴定信号蛋白复合物

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作者:Heng Lu, Qishan Lin, Jihe Zhao

Abstract

Protein-protein interactions play a pivotal role in both inter- and intra-cellular signaling. Identification of signaling protein complexes can thus shed important new insights into cell communications. We developed a parallel affinity precipitation protocol to overcome the disadvantages of the tandem affinity purification procedure, such as the potential disruption of target protein conformation, subcellular localization or function by epitope tags, the potential need of large amounts of cell culture or generation of stable cell lines, and relatively long duration the two-step precipitation takes. This new simplified assay of protein interaction is quick, economic and specific. This paper describes the details in the design and method of the assay.

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