A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells.

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.
Mesh Terms:
Animals, Binding Sites, Biotin, Biotinylation, Carbon-Nitrogen Ligases, Escherichia coli Proteins, HEK293 Cells, Humans, Laminin, Protein Interaction Mapping, Recombinant Fusion Proteins, Repressor Proteins
J. Cell Biol.
Date: Mar. 19, 2012
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