In vivo proximity labeling for the detection of protein-protein and protein-RNA interactions.

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 Res

authors

Beck DB,Narendra V,Drury WJ 3rd,Casey R,Jansen PW,Yuan ZF,Garcia BA,Vermeulen M,Bonasio R

doi

10.1021/pr500196b

subject

Has Abstract

pub_date

2014-12-05 00:00:00

pages

6135-43

issue

12

eissn

1535-3893

issn

1535-3907

journal_volume

13

pub_type

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