An improved smaller biotin ligase for BioID proximity labeling.
The BioID method uses a promiscuous biotin ligase to detect protein-protein associations as well as proximate proteins in living cells. Here we report improvements to the BioID method centered on BioID2, a substantially smaller promiscuous biotin ligase. BioID2 enables more-selective targeting of fusion proteins, requires less biotin supplementation, and exhibits ... enhanced labeling of proximate proteins. Thus BioID2 improves the efficiency of screening for protein-protein associations. We also demonstrate that the biotinylation range of BioID2 can be considerably modulated using flexible linkers, thus enabling application-specific adjustment of the biotin-labeling radius.
Mesh Terms:
Animals, Biotin, Biotinylation, Carbon-Nitrogen Ligases, Escherichia coli Proteins, Humans, Intracellular Signaling Peptides and Proteins, Membrane Proteins, Mice, Molecular Biology, NIH 3T3 Cells, Nuclear Pore Complex Proteins, Nuclear Proteins, Protein Engineering, Protein Interaction Mapping, Recombinant Fusion Proteins, Repressor Proteins
Animals, Biotin, Biotinylation, Carbon-Nitrogen Ligases, Escherichia coli Proteins, Humans, Intracellular Signaling Peptides and Proteins, Membrane Proteins, Mice, Molecular Biology, NIH 3T3 Cells, Nuclear Pore Complex Proteins, Nuclear Proteins, Protein Engineering, Protein Interaction Mapping, Recombinant Fusion Proteins, Repressor Proteins
Mol Biol Cell
Date: Apr. 15, 2016
PubMed ID: 26912792
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