Identification of Protein Networks Associated with the PAK1-PIX-GIT1-Paxillin Signaling Complex by Mass Spectrometry
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- 作者：Mark W. Mayhew ; Donna J. Webb ; Mykola Kovalenko ; Leanna Whitmore ; Jay W. Fox ; Alan F. Horwitz
- 刊名：Journal of Proteome Research
- 出版年：2006
- 出版时间：September 2006
- 年：2006
- 卷：5
- 期：9
- 页码：2417 - 2423
- 全文大小：329K
- 年卷期：v.5,no.9(September 2006)
- ISSN：1535-3907
文摘
The process of cell motility involves coordinate signaling events among proteins associated in interactiveintegrin-linked networks. Mass spectrometric analysis of immunoprecipitation-derived protein mixtureshave provided efficient means of identifying proteomes. In this study, we investigate strategies toenhance the detection of interactome proteins for the known signaling module: PAK1, 
PIX, GIT1, andpaxillin. Our results indicate that near-endogenous expression levels of bait protein enhances theidentification of associated proteins, and that phosphatase inhibition augments the detection of specificprotein interactions. Following the analysis of a large pool of spectral data, we have identified andmapped clusters of proteins that either share common interactions among the four bait proteins ofinterest or are exclusive to single bait proteins. Taken together, these data indicate that biochemicalmanipulations can enhance the ability for LC-MS/MS to identify interactome proteins, and thatqualitative screening of multiple samples leads to the compilation of proteins associated with a knownplexus.
PIX, GIT1, andpaxillin. Our results indicate that near-endogenous expression levels of bait protein enhances theidentification of associated proteins, and that phosphatase inhibition augments the detection of specificprotein interactions. Following the analysis of a large pool of spectral data, we have identified andmapped clusters of proteins that either share common interactions among the four bait proteins ofinterest or are exclusive to single bait proteins. Taken together, these data indicate that biochemicalmanipulations can enhance the ability for LC-MS/MS to identify interactome proteins, and thatqualitative screening of multiple samples leads to the compilation of proteins associated with a knownplexus.