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Literature summary extracted from

  • Howarth, M.; Ting, A.Y.
    Imaging proteins in live mammalian cells with biotin ligase and monovalent streptavidin (2008), Nat. Protoc., 3, 534-545.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
6.3.4.15 diagnostics BirA can be useful for specific in vivo labeling of proteins in cell cultures by biotinylation Escherichia coli

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.4.15 expression as His6-tagged enzyme in strain BL21(DE3) Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
6.3.4.15 additional information mammalian cell surface proteins tagged with a 15-amino acid peptide, i.e. the AP-tag, are specifically biotinylated by Escherichia coli biotin ligase BirA, whereas endogenous proteins are not modified, overview. BirA labels the AP-tag specifically in different cellular compartments: at the cell surface, in the endoplasmic reticulum, in the cytosol, and in the nucleus of mammalian cells, including human, mouse, rat and hamster cell lines, and primary neuron culture. BirA labeling may also be used in living animals, for example in a transgenic mouse expressing cytosolic BirA, overview. Benefit of streptavidin labeling over Ab labeling, method development, detailed overview Escherichia coli

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.4.15 Mg2+
-
 Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.4.15 ATP + biotin + apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)] Escherichia coli
-
 AMP + diphosphate + [acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.4.15 Escherichia coli
-
 gene birA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.4.15 recombinant His6-tagged enzyme from strain BL21(DE3) by nickel affinity chromatography Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.4.15 ATP + biotin + apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
-
 Escherichia coli AMP + diphosphate + [acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
-
 ?
6.3.4.15 additional information mammalian cell surface proteins tagged with a 15-amino acid peptide, i.e. the AP-tag, are specifically biotinylated by Escherichia coli biotin ligase BirA, whereas endogenous proteins are not modified, overview. BirA labels the AP-tag specifically in different cellular compartments: at the cell surface, in the endoplasmic reticulum, in the cytosol, and in the nucleus. BirA labeling may also be used in living animals, for example in a transgenic mouse expressing cytosolic BirA, overview Escherichia coli ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.3.4.15 biotin ligase
-
 Escherichia coli
6.3.4.15 BirA
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.4.15 37
-
 assay at Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.4.15 ATP
-
 Escherichia coli