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Anti BirA [9D11c1] Monoclonal Antibody

Invented by Brian Burke

Info

Catalogue Number 154118
Applications IF WB
Antigen/Gene or Protein Targets N-terminal half of E.coli BirA
Synonyms Anti-E.Coli BirA [9D11c1]
Reactivity E.coli
Host Mouse
Immunogen Glutathione-S-transferase fused to E.coli BirA R118G mutant
Immunogen UniProt ID P06709
Positive Control Cells overexpressing an E.coli BirA construct
Molecular Weight (kDa) 35kDa
Notes E.coli BirA is a well-characterized multifunctional protein that either catalyses the transfer of biotin to carboxyl carrier protein (BCCP), a acetyl-CoA carboxylase subunit or, alternatively, binds to the biotin operator resulting in transcriptional repression of the biotin operon (1). A mutant BirA (R118G) shows loss of DNA binding ability and promiscuity of substrate biotinylation (2, 3). These properties of BirA R118G were used to develop an in vivo technique that identifies proteins in a proximity- dependent manner (BioID). In the BioID method, a protein of interest is tagged with BirA R118G and expressed in live cells. Addition of the biotin in the media results in biotinylation of proteins in vicinity of the bait. The biotinylated proteins can then be affinity purified with biotin binding proteins such as strepavidin or avidin even under harsh denaturing conditions and subsequently identified by mass spectrometry or immunoblot analysis (4).

Reference
1. Streaker, E.D., D. Beckett. 2003. Coupling of protein assembly and DNA binding: biotin repressor dimerization precedes biotin operator binding. J Mol Biol. 325(5):937-48.
2. Kwon, K., E.D. Streaker, S. Ruparelia, D. Beckett. 2000. Multiple disordered loops function in corepressor-induced dimerization of the biotin repressor. J Mol Biol. 304(5):821-33.
3. Choi-Rhee, E., H. Schulman, J.E. Cronan. 2004. Promiscuous protein biotinylation by Escherichia coli biotin protein ligase. Protein Sci. 13(11):3043-50
4. Roux, K.J., D. I. Kim, M. Raida, B. Burke. 2012. A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells. J Cell Biol. 196(6):801-10.
Research Area Metabolism

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