Abstract:
:Accurate and sensitive detection of protein-protein and protein-RNA 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-protein and protein-RNA interactions in the nucleus of mammalian cells. Thus, IPL provides additional temporal and spatial information for the characterization of biological interactions in vivo.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Beck DB,Narendra V,Drury WJ 3rd,Casey R,Jansen PW,Yuan ZF,Garcia BA,Vermeulen M,Bonasio Rdoi
10.1021/pr500196bsubject
Has Abstractpub_date
2014-12-05 00:00:00pages
6135-43issue
12eissn
1535-3893issn
1535-3907journal_volume
13pub_type
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