In Vivo Proximity Labeling for the Detection of Protein鈥揚rotein and Protein鈥揜NA Interactions
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- 作者：David B. Beck ; Varun Narendra ; William J. Drury ; III ; Ryan Casey ; Pascal W. T. C. Jansen ; Zuo-Fei Yuan ; Benjamin A. Garcia ; Michiel Vermeulen ; Roberto Bonasio
- 刊名：Journal of Proteome Research
- 出版年：2014
- 出版时间：December 5, 2014
- 年：2014
- 卷：13
- 期：12
- 页码：6135-6143
- 全文大小：403K
- ISSN：1535-3907
文摘
Accurate and sensitive detection of protein鈥損rotein and protein鈥揜NA interactions is key to understanding their biological functions. Traditional methods to identify these interactions require cell lysis and biochemical manipulations that exclude cellular compartments that cannot be solubilized under mild conditions. Here, we introduce an in vivo proximity labeling (IPL) technology that employs an affinity tag combined with a photoactivatable probe to label polypeptides and RNAs in the vicinity of a protein of interest in vivo. Using quantitative mass spectrometry and deep sequencing, we show that IPL correctly identifies known protein鈥損rotein and protein鈥揜NA interactions in the nucleus of mammalian cells. Thus, IPL provides additional temporal and spatial information for the characterization of biological interactions in vivo.