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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.

SUBMITTER: Beck DB

PROVIDER: S-EPMC4261942 | biostudies-literature | 2014 Dec

REPOSITORIES: biostudies-literature

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

Beck David B DB Narendra Varun V Drury William J WJ Casey Ryan R Jansen Pascal W T C PW Yuan Zuo-Fei ZF Garcia Benjamin A BA Vermeulen Michiel M Bonasio Roberto R

Journal of proteome research 20141029 12

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 o ...[more]

PMID: 25311790

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