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J Cell Biol. 2012 Mar 19;196(6):801-10. doi: 10.1083/jcb.201112098. Epub 2012 Mar 12.

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

Author information

1
Sanford Children's Health Research Center, Sanford Research/University of South Dakota, Sioux Falls, SD 57104, USA. Kyle.Roux@sanfordhealth.org

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.

PMID:
22412018
PMCID:
PMC3308701
DOI:
10.1083/jcb.201112098
[Indexed for MEDLINE]
Free PMC Article

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