What is BirA protein?
BirA (bifunctional biotin-[acetylCoA carboxylase] holoenzyme synthetase/ DNA-binding transcriptional repressor, bio-5′-AMP-binding) is a bifunctional protein that exhibits biotin ligase activity and also acts as the DNA binding transcriptional repressor of the biotin operon.
What is BirA ligase?
BirA is the Escherichia coli biotin ligase that site-specifically biotinylates a lysine side chain within a 15-amino acid acceptor peptide (also known as Avi-tag).
What is BirA enzyme?
The enzyme BirA is a key reagent because of its ability to biotinylate proteins at a specific residue in a recognition sequence. We report a rapid, efficient, and economical method for the production, purification, and application of this enzyme.
What is the function of BirA?
In certain bacteria, such as Escherichia coli and Staphylococcus aureus, BirA is a bi-functional protein that serves as a transcriptional repressor to regulate biotin biosynthesis genes, as well as acting as a ligase to catalyze the biotinylation of biotin-dependent enzymes.
How does AviTag work?
Using AviTag, one can label a protein of interest with biotin by simply creating a fusion in which AviTag is added to either the N or C terminus of the protein. BirA has been shown to effectively biotinylate AviTag expressing proteins in bacteria, yeast, insect and mammalian cells.
How do you Biotinylate a protein?
coli biotin holoenzyme synthetase, also known as biotin ligase (BirA, P06709). The most common way of targeting a protein of interest is by fusing the protein at its N-terminus, C-terminus or at an internal loop to a 15 amino acid peptide ( GLNDIFEAQKIEWHE ), termed AviTag or Acceptor Peptide (AP).
What is biotin ligase?
Biotin-protein ligase (EC 6.3. 4.15) activates biotin to form biotinyl 5′ adenylate and transfers the biotin to biotin-accepting proteins. It also functions as a biotin operon repressor. The protein is encoded by the birA gene. Biomix A, B, and d-biotin can be stored at -20°C.
How do you take BioID?
The following steps outline the BioID process. (1) Ligate bait cDNA and BioID in-frame and in an appropriate expression vector. (2) Express BioID fusion protein in cells to (3) generate stably expressing cells. (4) Induce biotinylation with excess biotin prior to (5) cell lysis and protein denaturation.
Which amino acids can be biotinylated?
Chemical structure of biotin. The molecule is comprised of a ureido ring joined with a tetrahydrothiophene ring. A coenzyme for carboxylase enzymes, biotin is required for the synthesis of fatty acids, isoleucine and valine.
What is Bira white?
Product description. BIRA White Ale is a deliciously different wheat beer with low bitterness, a hint of spicy citrus with a soft finish – a refreshing all day craft beer, brewed with the finest natural ingredients mixed with barrels of passion.
How do you tag proteins?
Tagging can be done via cloning into vectors or added using CRISPR-Cas9 gene editing to tag an endogenous protein. By using an affinity tag, you can isolate or immobilize a protein for additional proteomic studies.
What is SUMO tag?
SUMO Tag Definition Sumo tag is most frequently used as N-end fusion sequence in yeast to increase the expression and solubility of the desired recombinant protein. SUMO proteins are similar to ubiquitin in their folded structure but possess only about 20% homology to the amino acid sequence of ubiquitin.
What kind of enzyme is the Bira gene?
Our BirA enzyme is E. coli wild-type, encoded by the birA gene, and purified to greater than 99% purity by traditional methods.
How is Bira used for site specific biotinylation?
Enzymatic biotinylation with E. coli biotin ligase (BirA) is highly specific in covalently attaching biotin to the 15 amino acid AviTag peptide, giving a homogeneous product with high yield. AviTag can conveniently be added genetically at the N-terminus, C-terminus or in exposed loops of a target protein.
What is the function of Bira biotin protein ligase?
BirA Biotin-protein ligase (EC 188.8.131.52) activates biotin to form biotinyl 5′-adenylate and transfers the biotin to biotin-accepting proteins (such as AviTag™ peptide). It also functions as a biotin operon repressor.
How does Bira regulate the activity of acetyl-CoA?
In E. coli and other bacteria, BirA regulates the enzyme activity of acetyl-CoA carboxylase through biotinylation of the carboxyl carrier protein (BCCP) subunit . BirA also helps coordinate biotin synthesis by regulating transcription of the biotin biosynthetic operon , , .