6.3.4.15: biotin-[biotin carboxyl-carrier protein] ligase
This is an abbreviated version!
For detailed information about biotin-[biotin carboxyl-carrier protein] ligase, go to the full flat file.
Word Map on EC 6.3.4.15
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6.3.4.15
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biotinylation
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streptavidin
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bioid
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proximity-dependent
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bap
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avidin
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avitag
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streptavidin-coated
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biotin-dependent
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transcarboxylase
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proximity-labeling
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biotinyl-5\'-amp
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drug development
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unbiotinylated
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15-amino
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biotin-streptavidin
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diagnostics
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analysis
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synthesis
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carboxylases
- 6.3.4.15
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biotinylation
- streptavidin
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bioid
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proximity-dependent
- bap
- avidin
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avitag
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streptavidin-coated
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biotin-dependent
- transcarboxylase
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proximity-labeling
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biotinyl-5\'-amp
- drug development
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unbiotinylated
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15-amino
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biotin-streptavidin
- diagnostics
- analysis
- synthesis
- carboxylases
Reaction
Synonyms
Acetyl CoA holocarboxylase synthetase, Acetyl coenzyme A holocarboxylase synthetase, bacterial BirA biotin ligase, biotin acetyl-CoA carboxylase ligase, Biotin holoenzyme synthetase, biotin ligase, biotin protein ligase, Biotin--protein ligase, biotin-protein ligase, Biotin-[acetyl coenzyme A carboxylase] synthetase, Biotin-[acetyl-CoA carboxylase] synthetase, Biotin:apocarboxylase ligase, BirA, BirA protein, BPL, group I biotin protein ligase, HCS, Holocarboxylase synthetase, holocarboxylase synthetase 1, More, STK_15250, Synthetase, biotin-[acetyl coenzyme A carboxylase], yBL
ECTree
6.3.4.15 biotin-[biotin carboxyl-carrier protein] ligaseAdvanced search results
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results (Engineering
Engineering on EC 6.3.4.15 - biotin-[biotin carboxyl-carrier protein] ligase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
R40G
site-directed mutagenesis, the structure of the mutant enzyme in both the ligand-free and biotin-bound forms reveals that the mutated loop has collapsed, thus hindering ATP binding. The mutant is catalytically active but shows poor substrate specificity. The affinity for biotin is reduced 3.5fold by the R40G mutation, but binding of ATP to mutant R40G is very weak in the absence or in the presence of biotin. Comparison to the equivalent mutant of Escherichia coli BirA, overview
DELTA1-81
elimination of N-terminal domain plus part of the central domain, loss of ligase activity
DELTA2-63
elimination of N-terminal domain, mutant shows normal ligase activity
DELTA2-65
elimination of N-terminal domain, mutant shows normal ligase activity
DELTA1-81
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elimination of N-terminal domain plus part of the central domain, loss of ligase activity
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DELTA2-63
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elimination of N-terminal domain, mutant shows normal ligase activity
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DELTA2-65
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elimination of N-terminal domain, mutant shows normal ligase activity
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G115S
I11V/P85S/ M157T/K163E/I224T
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17fold increase in the ability to use desthiobiotin as a substrate
L237P
the mutant shows reduced catalytic activity compared to the wild-type enzyme
Q41R/M157T/I224T/E215D/T225A
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10fold increase in the ability to use desthiobiotin as a substrate
R118G
D571N
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naturally occuring mutation, important in positioning K579 in the AMP binding site
D615Y
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naturally occuring mutation in the loop between alpha3 and beta6 that cover AMP, may coordinate oxygens of the AMP phosphate
D634Y
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naturally occuring mutation of a solvent exposed residue, distal to active site on alpha4 and alpha5, respectively
D715G
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naturally occuring mutation on beta9, may be involved in capping and stabilising the catalytic domain structure
G518E
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naturally occuring mutation close to the active site and part of the ligand-binding loop, the mutation may not allow as much flexibility of this loop. As part of the hydrophobic pocket, making this polar residue would perturb biotin binding
G581S
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naturally occuring mutation of a residue involved in hydrophobic interactions with biotin in the binding pocket
G582R
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naturally occuring mutation of a residue involved in hydrophobic interactions with biotin in the binding pocket
L470S
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naturally occuring mutation in an unstructured loop distal to the ligand-binding site
R508W/N511K
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naturally occuring mutation of a residue critical in loop covering the ligand-binding site. R508 co-ordinates to the backbone carbonyl N712 to form a salt bridge, removing this would result in a more flexible loop, analogous to EcBPL R118 and R121 that co-ordinate oxygens within the AMP phosphate group
T462I
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naturally occuring mutation in an unstructured loop distal to the ligand-binding site
V547G
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naturally occuring mutation of a buried hydrophobic residue in alpha2 that reside near the AMP-binding beta5 strand
V550M
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naturally occuring mutation of a buried hydrophobic residue in alpha2 that reside near the AMP-binding beta5 strand
Y456C
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naturally occuring mutation in an unstructured loop distal to the ligand-binding site
R69A
the binding constant for biotin is nearly the same as that observed for the wild type protein. Mutant does not undergo self-botinylation
R48A/K111A
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site-directed mutagenesis, comparison of the nucleotide binding mode in the double mutant to that in the wild-type enzyme, the active site loop Gly45-Trp53 is disordered in the mutant. Interface charge distribution in the double mutant complex, overview
R48K
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site-directed mutagenesis, comparison of the nucleotide binding mode in the mutant to that in the wild-type enzyme, the active site loop Gly45-Trp53 is disordered in the mutant
R48A/K111A
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site-directed mutagenesis, comparison of the nucleotide binding mode in the double mutant to that in the wild-type enzyme, the active site loop Gly45-Trp53 is disordered in the mutant. Interface charge distribution in the double mutant complex, overview
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R48K
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site-directed mutagenesis, comparison of the nucleotide binding mode in the mutant to that in the wild-type enzyme, the active site loop Gly45-Trp53 is disordered in the mutant
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DELTA1-81
elimination of N-terminal domain plus part of the central domain, complete loss of ligase activity
DELTA2-65
elimination of N-terminal domain, complete loss of ligase activity
DELTA2-74
elimination of N-terminal domain plus part of the central domain, complete loss of ligase activity
F123G
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the mutation has minimal effect upon the Km value for MgATP and biotin. In contrast, the kcat for biotin and MgATP are significantly compromised to about 10% that of wild type enzyme
F123R
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the mutation has minimal effect upon the Km value for MgATP and biotin. In contrast, the kcat for biotin and MgATP are significantly compromised to about 10% that of wild type enzyme
R125A
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the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme
R125N
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the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme
R125A
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the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme
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R125N
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the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme
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F123G
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the mutation has minimal effect upon the Km value for MgATP and biotin. In contrast, the kcat for biotin and MgATP are significantly compromised to about 10% that of wild type enzyme
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F123R
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the mutation has minimal effect upon the Km value for MgATP and biotin. In contrast, the kcat for biotin and MgATP are significantly compromised to about 10% that of wild type enzyme
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DELTA1-81
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elimination of N-terminal domain plus part of the central domain, complete loss of ligase activity
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DELTA2-65
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elimination of N-terminal domain, complete loss of ligase activity
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DELTA2-74
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elimination of N-terminal domain plus part of the central domain, complete loss of ligase activity
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A43G
additional information
G115S
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site-directed mutagenesis, the mutant enzyme shows a highly increased rate of biotinyl-5'-AMP intermediate formation, but no release of the intermediate
R118G
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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
R118G
mutation of Arg118 to glycine in EcBPL causes altered affinity of BPL for biotin, increased dissociation rates for biotin and biotinyl-5'-AMP, and consequently, promiscuous biotinylation of inappropriate proteins
A43G
catalytic efficiency is 2fold lower than catalytic efficiency of wild-type enzyme
A43G
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catalytic efficiency is 2fold lower than catalytic efficiency of wild-type enzyme
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additional information
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disruption of BPL affects important metabolic pathways such as fatty acid biosynthesis and gluconeogenesis
additional information
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disruption of BPL affects important metabolic pathways such as fatty acid biosynthesis and gluconeogenesis
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additional information
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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
additional information
mutational analysis, overview. The C-terminal 87 amino acids of biotin carboxyl carrier protein, i.e. BCCP-87, are recognised and biotinylated by BPL as well as full-length BCCP due to the fact that this peptide adopts the tertiary structure that is required for the interaction with BPL
additional information
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HCS mutants with deletions up to Ala235 or Thr266 still show catalytic activity and can complement an enzyme-deficient Escherichia coli birA- strain. Enzyme deficiency can cause symptoms like ketoacidosis, feeding difficulties, hypotonia, seizures, developmental delay and dermal abnormalities such as rashes, dryness of the skin and alopecia, multiple carboxylase deficiency is caused by a lack of activity of the biotin-dependent enzymes, phenotypes, overview. Molecular modelling of mutations causing HCS deficiency, detailed overview
additional information
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construction of biotinylated magnetic nanoparticles, with or without introduced strepatavidin, by displaying biotin acceptor peptide or biotin carboxyl carrier protein, BCCP, on the surface of bacterial magnetic particles for analysis of in vitro biotinylation, method development and evaluation, overview
additional information
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construction of biotinylated magnetic nanoparticles, with or without introduced strepatavidin, by displaying biotin acceptor peptide or biotin carboxyl carrier protein, BCCP, on the surface of bacterial magnetic particles for analysis of in vitro biotinylation, method development and evaluation, overview
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