title: An improved smaller biotin ligase for BioID proximity labeling.
abstract: 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.
authors.affiliation: Sanford Children's Health Research Center, Sanford Research, Sioux Falls, SD 57104.
authors.forename: Dae In
authors.initials: DI
authors.lastname: Kim
authors.affiliation: Sanford Children's Health Research Center, Sanford Research, Sioux Falls, SD 57104.
authors.forename: Samuel C
authors.initials: SC
authors.lastname: Jensen
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306.
authors.forename: Kyle A
authors.initials: KA
authors.lastname: Noble
authors.affiliation: Sanford Children's Health Research Center, Sanford Research, Sioux Falls, SD 57104.
authors.forename: Birendra
authors.initials: B
authors.lastname: Kc
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306.
authors.forename: Kenneth H
authors.initials: KH
authors.lastname: Roux
authors.affiliation: Sanford-Burnham Proteomics Facility, Sanford-Burnham Medical Research Institute, La Jolla, CA 92037.
authors.forename: Khatereh
authors.initials: K
authors.lastname: Motamedchaboki
authors.affiliation: Sanford Children's Health Research Center, Sanford Research, Sioux Falls, SD 57104 Department of Pediatrics, Sanford School of Medicine, University of South Dakota, Sioux Falls, SD 57105 kyle.roux@sanfordhealth.org.
authors.forename: Kyle J
authors.initials: KJ
authors.lastname: Roux
mesh_terms: D000818:Animals; D001710:Biotin / Q000378:metabolism; D019921:Biotinylation; D019731:Carbon-Nitrogen Ligases / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D041681:NIH 3T3 Cells; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods; D025941:Protein Interaction Mapping / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C512772:NUP107 protein, human; C440251:NUP160 protein, human; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C413025:SUN2 protein, human; D001710:Biotin; D019731:Carbon-Nitrogen Ligases; C058125:birA protein, E coli