An improved smaller biotin ligase for BioID proximity labeling

Dae In Kim; Samuel C Jensen; Kyle A Noble; Birendra Kc; Kenneth H Roux; Khatereh Motamedchaboki; Kyle J Roux

doi:10.1091/mbc.E15-12-0844

An improved smaller biotin ligase for BioID proximity labeling

Mol Biol Cell. 2016 Apr 15;27(8):1188-96. doi: 10.1091/mbc.E15-12-0844. Epub 2016 Feb 24.

Authors

Dae In Kim¹, Samuel C Jensen¹, Kyle A Noble², Birendra Kc¹, Kenneth H Roux², Khatereh Motamedchaboki³, Kyle J Roux⁴

Affiliations

¹ Sanford Children's Health Research Center, Sanford Research, Sioux Falls, SD 57104.
² Department of Biological Science, Florida State University, Tallahassee, FL 32306.
³ Sanford-Burnham Proteomics Facility, Sanford-Burnham Medical Research Institute, La Jolla, CA 92037.
⁴ 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.

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.

© 2016 Kim et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Biotin / metabolism
Biotinylation
Carbon-Nitrogen Ligases / genetics
Carbon-Nitrogen Ligases / metabolism*
Escherichia coli Proteins / genetics
Escherichia coli Proteins / metabolism*
Humans
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism
Membrane Proteins / genetics
Membrane Proteins / metabolism
Mice
Molecular Biology / methods*
NIH 3T3 Cells
Nuclear Pore Complex Proteins / genetics
Nuclear Pore Complex Proteins / metabolism
Nuclear Proteins / genetics
Nuclear Proteins / metabolism
Protein Engineering / methods
Protein Interaction Mapping / methods
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism*
Repressor Proteins / genetics
Repressor Proteins / metabolism*

Substances

Escherichia coli Proteins
Intracellular Signaling Peptides and Proteins
Membrane Proteins
NUP107 protein, human
NUP160 protein, human
Nuclear Pore Complex Proteins
Nuclear Proteins
Recombinant Fusion Proteins
Repressor Proteins
SUN2 protein, human
Biotin
Carbon-Nitrogen Ligases
birA protein, E coli

Abstract

Publication types

MeSH terms

Substances

Grants and funding