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PMID:22412018
| Citation |
Roux, KJ, Kim, DI, Raida, M and Burke, B (2012) A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells. J. Cell Biol. 196:801-10 |
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| Abstract |
We have developed a new technique for proximity-dependent labeling of proteins in eukaryotic cells. Named BioID for proximity-dependent biotin identification, this approach is based on fusion of a promiscuous Escherichia coli biotin protein ligase to a targeting protein. BioID features proximity-dependent biotinylation of proteins that are near-neighbors of the fusion protein. Biotinylated proteins may be isolated by affinity capture and identified by mass spectrometry. We apply BioID to lamin-A (LaA), a well-characterized intermediate filament protein that is a constituent of the nuclear lamina, an important structural element of the nuclear envelope (NE). We identify multiple proteins that associate with and/or are proximate to LaA in vivo. The most abundant of these include known interactors of LaA that are localized to the NE, as well as a new NE-associated protein named SLAP75. Our results suggest BioID is a useful and generally applicable method to screen for both interacting and neighboring proteins in their native cellular environment. |
| Links |
PubMed PMC3308701 Online version:10.1083/jcb.201112098 |
| Keywords |
Animals; Binding Sites; Biotin/metabolism; Biotinylation; Carbon-Nitrogen Ligases/genetics; Carbon-Nitrogen Ligases/metabolism; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; HEK293 Cells; Humans; Laminin/genetics; Laminin/metabolism; Protein Interaction Mapping/methods; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Repressor Proteins/genetics; Repressor Proteins/metabolism |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0009305: protein biotinylation |
ECO:0000314: |
P |
Figure 2 shows that "BirA* promiscuously biotinylates endogenous proteins in mammalian cells." |
complete | ||||
See also
References
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