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Nat Commun. 2017 Jun 6;8:15690. doi: 10.1038/ncomms15690.

Split-BioID a conditional proteomics approach to monitor the composition of spatiotemporally defined protein complexes.

Author information

1
CellNetworks Junior Research Group Posttranscriptional Regulation of mRNA Expression and Localization, Heidelberg University, INF 328, D-69120 Heidelberg, Germany.
2
Biochemie-Zentrum Heidelberg (BZH), INF 328, D-69120 Heidelberg, Germany.

Abstract

Understanding the function of the thousands of cellular proteins is a central question in molecular cell biology. As proteins are typically part of multiple dynamic and often overlapping macromolecular complexes exerting distinct functions, the identification of protein-protein interactions (PPI) and their assignment to specific complexes is a crucial but challenging task. We present a protein fragments complementation assay integrated with the proximity-dependent biotinylation technique BioID. Activated on the interaction of two proteins, split-BioID is a conditional proteomics approach that allows in a single and simple assay to both experimentally validate binary PPI and to unbiasedly identify additional interacting factors. Applying our method to the miRNA-mediated silencing pathway, we can probe the proteomes of two distinct functional complexes containing the Ago2 protein and uncover the protein GIGYF2 as a regulator of miRNA-mediated translation repression. Hence, we provide a novel tool to study dynamic spatiotemporally defined protein complexes in their native cellular environment.

PMID:
28585547
PMCID:
PMC5467174
DOI:
10.1038/ncomms15690
[Indexed for MEDLINE]
Free PMC Article

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