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Literature summary for 6.3.4.15 extracted from

  • Choi-Rhee, E.; Schulman, H.; Cronan, J.E.
    Promiscuous protein biotinylation by Escherichia coli biotin protein ligase (2004), Protein Sci., 13, 3043-3050.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.3.4.15

Cloned(Commentary)

Cloned (Comment) Organism
gene birA, expression of His-tagged wild-type and mutant enzymes in strain JM109, expression of enzyme in strain BM4092 unbiotinylated form Escherichia coli

Protein Variants

Protein Variants Comment Organism
G115S site-directed mutagenesis, the mutant enzyme shows a highly increased rate of biotinyl-5'-AMP intermediate formation, but no release of the intermediate Escherichia coli
R118G site-directed mutagenesis, the mutant enzyme shows a wider substrate specificity compared to the wild-type enzyme, the mutant biotinylates bovine serum albumin, chloramphenicol acetyltransferase, immunoglobin heavy and light chains, and RNase A in vitro, it also self-biotinylates in vivo and in vitro, the activity depends on proximity, distance to the substrate can be bridged by a linker like avidin, overview Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
 Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from strain JM109 by nickel affinity chromatography Escherichia coli

Reaction

Reaction Comment Organism Reaction ID
ATP + biotin + [biotin carboxyl-carrier protein]-L-lysine = AMP + diphosphate + [biotin carboxyl-carrier protein]-N6-biotinyl-L-lysine reaction mechanism, the first step is the reversible formation of a biotinyl-5'-AMP intermediate followed by irreversible transfer of the biotinyl group to the protein Escherichia coli
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information substrate specificities of wild-type enzyme and of mutant R118G, the first biotinylates only a single protein substrate, while the mutant shows a wider substrate spectrum Escherichia coli ?
-
 ?

Synonyms

Synonyms Comment Organism
biotin protein ligase
-
 Escherichia coli
BirA
-
 Escherichia coli
BPL
-
 Escherichia coli

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Escherichia coli