Information on EC 6.3.4.15 - biotin-[acetyl-CoA-carboxylase] ligase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
6.3.4.15
-
RECOMMENDED NAME
GeneOntology No.
biotin-[acetyl-CoA-carboxylase] ligase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + biotin + apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)] = AMP + diphosphate + [acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
amination
-
-
-
-
carboxylic acid formation
-
-
carboxamide formation
-
additional information
biotinylation, mediates attachment of biotin to a fusion protein of a biotin acceptor peptide and GLuc, EC 1.13.12.5
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biotin metabolism
-
-
biotin-carboxyl carrier protein assembly
-
-
Metabolic pathways
-
-
biotin biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
biotin:apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)] ligase (AMP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY hide
37340-95-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain CBF562
-
-
Manually annotated by BRENDA team
strain CBF562
-
-
Manually annotated by BRENDA team
strain AMB-1
-
-
Manually annotated by BRENDA team
strain AMB-1
-
-
Manually annotated by BRENDA team
biotin-deficient animals
-
-
Manually annotated by BRENDA team
-
E5R5T0
UniProt
Manually annotated by BRENDA team
-
E5R5T0
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + biotin + apo-[acetyl CoA carboxylase]
AMP + diphosphate + acetyl CoA carboxylase
show the reaction diagram
no coupling is observed in ATP and biotin binding to the BPL
-
-
?
ATP + biotin + apo-[acetyl-CoA carboxylase 1]
AMP + diphosphate + acetyl-CoA carboxylase 1
show the reaction diagram
-
cytoplasmic acetyl-CoA carboxylase isozyme
-
-
?
ATP + biotin + apo-[acetyl-CoA carboxylase 2]
AMP + diphosphate + acetyl-CoA carboxylase 2
show the reaction diagram
ATP + biotin + apo-[acetyl-CoA carboxylase]
AMP + diphosphate + acetyl-CoA carboxylase
show the reaction diagram
ATP + biotin + apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
AMP + diphosphate + [acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
show the reaction diagram
ATP + biotin + apo-[biotin carboxyl carrier protein]
AMP + diphosphate + biotin carboxyl carrier protein
show the reaction diagram
-
i.e. BCCP, substrate is the recombinantly expressed His-tagged biotinoyl domain, BCCP87, of Escherichia coli BCCP, binding pattern, overview
-
-
?
ATP + biotin + apo-[biotin carboxyl carrier protein]
AMP + diphosphate + biotinylated biotin carboxyl carrier protein
show the reaction diagram
ATP + biotin + apo-[propionyl-CoA carboxylase]
AMP + diphosphate + propionyl-CoA carboxylase
show the reaction diagram
-
-
-
-
?
ATP + biotin + apocarboxylase
AMP + diphosphate + holocarboxylase
show the reaction diagram
-
-
-
-
?
ATP + biotin + Avi-tagged thioredoxin
?
show the reaction diagram
ATP + biotin + C-terminal domain of apo-biotin carboxyl carrier protein
AMP + diphosphate + biotinylated C-terminal domain of acetyl CoA carboxylase
show the reaction diagram
-
Escherichia coli BPL biotinylates both the homologous BCCP domain BCCP87 and the corresponding domain from Mycobacterium tuberculosis
-
-
?
ATP + biotin + C-terminal domain of apo-biotin carboxyl carrier protein
AMP + diphosphate + biotinylated C-terminal domain of apo-biotin carboxyl carrier protein
show the reaction diagram
-
Mycobacterium tuberculosis BPL specifically biotinylates the homologous BCCP domain BCCP87, but not the Escherichia coli domain BCCP87
-
-
?
ATP + biotin + carboxyl carrier protein
?
show the reaction diagram
-
the biotinylated product, holo biotin carboxyl carrier protein, forms a stable complex with the enzyme, which barely dissociates at room temperature
-
-
?
ATP + biotin + chimeric protein of glutathione S-transferase and green fluorescence protein with biotin carboxyl carrier proteinDelta100
?
show the reaction diagram
-
-
-
-
?
ATP + biotin + fluorescein-labeled peptide 85-11
ADP + diphosphate + biotinyl-fluorescein-labeled peptide 85-11
show the reaction diagram
-
peptide 85-11 has the sequence MAGGLNDIFEAQKIEWHE with the lysine residue being targeted for biotinylation, development of a fluorescence polarization technology-based assay method, overview
-
-
?
ATP + biotin + GLNDIFEAQKIEWH
AMP + diphosphate + biotinylated GLNDIFEAQKIEWH
show the reaction diagram
-
i.e. Schatz' peptide, synthetic biotinable minimal peptide
-
-
?
ATP + biotin + protein p67
AMP + diphosphate + biotinyl-protein p67
show the reaction diagram
ATP + biotin + yeast acceptor peptide
?
show the reaction diagram
-
-
-
-
?
ATP + biotin + [Escherichia coli apo-biotin-carboxyl-carrier-protein mutant M136S]-L-lysine
AMP + diphosphate + [Escherichia coli biotin-carboxyl-carrier-protein mutant M136S]-biotinyl-L-lysine
show the reaction diagram
-
biotin carboxyl carrier protein (BCCP) is one subunit or domain of biotin-dependent enzymes. Biotin carboxyl carrier protein becomes an active substrate for carboxylation and carboxyl transfer, after biotinylation of its canonical lysine residue by biotin protein ligase (BPL). BCCP carries a characteristic local sequence surrounding the canonical lysine residue, typically -M-K-M-. Sulfolobus tokodaii is unique in that its biotin carboxyl carrier protein has serine replaced for the methionine C-terminal to the lysine. This biotin carboxyl carrier protein is biotinylated by its own biotin protein ligase, but not by Escherichia coli biotin protein ligase. Likewise, Escherichia coli biotin carboxyl carrier protein is not biotinylated by Sulfolobus tokodaii biotin protein ligase
-
-
?
ATP + biotin + [Escherichia coli apo-biotin-carboxyl-carrier-protein]-L-lysine
AMP + diphosphate + [Escherichia coli biotin-carboxyl-carrier-protein]-biotinyl-L-lysine
show the reaction diagram
-
biotin carboxyl carrier protein (BCCP) is one subunit or domain of biotin-dependent enzymes. Biotin carboxyl carrier protein becomes an active substrate for carboxylation and carboxyl transfer, after biotinylation of its canonical lysine residue by biotin protein ligase (BPL). BCCP carries a characteristic local sequence surrounding the canonical lysine residue, typically -M-K-M-. Sulfolobus tokodaii is unique in that its biotin carboxyl carrier protein has serine replaced for the methionine C-terminal to the lysine. This biotin carboxyl carrier protein is biotinylated by its own biotin protein ligase, but not by Escherichia coli biotin protein ligase. Likewise, Escherichia coli biotin carboxyl carrier protein is not biotinylated by Sulfolobus tokodaii biotin protein ligase
-
-
?
ATP + biotin + [Sulfolobus tokodaii apo-biotin-carboxyl-carrier-protein mutant S136M]-lysine
AMP + diphosphate + [Sulfolobus tokodaii biotin-carboxyl-carrier-protein mutant S136M]-biotinyllysine
show the reaction diagram
ATP + biotin + [Sulfolobus tokodaii apo-biotin-carboxyl-carrier-protein]-lysine
AMP + diphosphate + [Sulfolobus tokodaii biotin-carboxyl-carrier-protein]-biotinyllysine
show the reaction diagram
ATP + cis-propargyl biotin + apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
AMP + diphosphate + cis-propargyl bioti-containing [acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
show the reaction diagram
-
-
-
-
?
ATP + cis-propargyl biotin + apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
AMP + diphosphate + cis-propargyl biotin-containing [acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
show the reaction diagram
-
-
-
-
?
ATP + desthiobiotin azide + apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
AMP + diphosphate + desthiobiotin azide-containing [acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
show the reaction diagram
-
-
-
-
?
avitag-tagged maltose-binding protein + biotin + ATP
maltose-binding protein-avitag-biotin + AMP + diphosphate
show the reaction diagram
avitagged BirA + biotin + ATP
BirA-avitag-biotin + AMP + diphosphate
show the reaction diagram
-
recombinant Saccharomyces cerevisiae cells
-
-
?
avitagged BirA + biotin + biotin + ATP
BirA-avitag-biotin + AMP + diphosphate
show the reaction diagram
-
the recombinant avitagged enzyme biotinylates itself at the avitag sequence
-
-
?
biotin + ATP
biotinyl-5'-AMP + diphosphate
show the reaction diagram
-
first half-reaction of BPL
-
-
?
biotin + ATP + apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
AMP + diphosphate + [acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
show the reaction diagram
-
catalyzes the covalent attachment of the biotin prosthetic group to a specific lysine of the biotin carboxyl carrier domain of biotin-dependent carboxylases in a two-step reaction
-
r
biotinyl-5'-AMP + apocarboxylase
holocarboxylase + AMP
show the reaction diagram
D-biotin + ATP
biotinyl-5'-AMP + diphosphate
show the reaction diagram
-
first half-reaction of BPL
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + biotin + apo-[acetyl-CoA carboxylase 1]
AMP + diphosphate + acetyl-CoA carboxylase 1
show the reaction diagram
-
cytoplasmic acetyl-CoA carboxylase isozyme
-
-
?
ATP + biotin + apo-[acetyl-CoA carboxylase 2]
AMP + diphosphate + acetyl-CoA carboxylase 2
show the reaction diagram
-
mitochondrial acetyl-CoA carboxylase isozyme
-
-
?
ATP + biotin + apo-[acetyl-CoA carboxylase]
AMP + diphosphate + acetyl-CoA carboxylase
show the reaction diagram
ATP + biotin + apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
AMP + diphosphate + [acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
show the reaction diagram
ATP + biotin + apocarboxylase
AMP + diphosphate + holocarboxylase
show the reaction diagram
-
-
-
-
?
avitag-tagged maltose-binding protein + biotin + ATP
maltose-binding protein-avitag-biotin + AMP + diphosphate
show the reaction diagram
-
recombinant Saccharomyces cerevisiae cells
-
-
?
avitagged BirA + biotin + ATP
BirA-avitag-biotin + AMP + diphosphate
show the reaction diagram
-
recombinant Saccharomyces cerevisiae cells
-
-
?
biotin + ATP
biotinyl-5'-AMP + diphosphate
show the reaction diagram
-
first half-reaction of BPL
-
-
?
biotinyl-5'-AMP + apocarboxylase
holocarboxylase + AMP
show the reaction diagram
-
second half-reaction of BPL, the apocarboxylase is the biotin-carboxyl-carrier protein, which is carboxylated after biotin binding by the biotin carboxylase, BCCP, EC 6.3.4.14
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
biotin
cis-propargyl biotin
desthiobiotin azide
-
can substitute for biotin
additional information
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(3aS,4S,6aR)-4-(5-[1-[3-(naphthalen-2-yloxy)propyl]-1H-1,2,3-triazol-4-yl]pentyl)tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one
-
-
(3aS,4S,6aR)-4-(5-[1-[4-(naphthalen-1-yloxy)butyl]-1H-1,2,3-triazol-4-yl]pentyl)tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one
-
-
(3aS,4S,6aR)-4-(5-[1-[4-(naphthalen-2-yloxy)butyl]-1H-1,2,3-triazol-4-yl]pentyl)tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one
-
-
(3aS,4S,6aR)-4-[-[1-[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl]methyl]triazol-4-yl]pentyl]-1,3,3a,4,6,6ahexahydrothieno[3,4-d]imidazol-2-one
-
competitive inhibitor
(3aS,4S,6aR)-4-[4-[1-[[(3aR,4R,6R,6aR)-4-(6-aminopurin-9-yl)-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-6-yl]methyl]triazol-4-yl]butyl]-1,3,3a,4,6,6a-hexa hydrothieno[3,4-d]imidazol-2-one
-
-
(3aS,4S,6aR)-4-[5-[1-(4-phenoxybutyl)-1H-1,2,3-triazol-4-yl]pentyl]tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one
-
-
(3aS,4S,6aR)-4-[5-[1-[[(3aR,4R,6R,6aR)-4-(6-aminopurin-9-yl)-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-6-yl]methyl]triazol-4-yl]pentyl]-1,3,3a,4,6,6a-hexa hydrothieno[3,4-d]imidazol-2-one
-
-
(3aS,4S,6aR)-4-[5-[3-[[(3aR,4R,6R,6aR)-4-(6-aminopurin-9-yl)-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-6-yl]methyl]triazol-4-yl]pentyl]-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-2-one
-
-
(3aS,4S,6aR)-4-[6-[1-[[(3aR,4R,6R,6aR)-4-(6-aminopurin-9-yl)-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-6-yl]methyl]triazol-4-yl]hexyl]-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-2-one
-
-
2-(4-[5-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentyl]-1H-1,2,3-triazol-1-yl)ethyl 2,2-dimethylpropanoate
-
-
5-methyl-3-[3-(4-[5-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentyl]-1H-1,2,3-triazol-1-yl)propyl]-1,3-benzoxazol-2(3H)-one
-
-
5-methyl-3-[4-(4-[5-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentyl]-1H-1,2,3-triazol-1-yl)butyl]-1,3-benzoxazol-2(3H)-one
-
competitive inhibitor versus biotin
biotin acetylene
-
i.e. (3aS,4S,6aR)-4-hept-6-ynyl-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-2-one
biotinol-5'-AMP
-
-
biotinyl-5'-AMP
-
selective inhibitor
diphosphate
-
endproduct inhibition
GLNDIFEAQKIEWH
-
i.e. Schatz' peptide, synthetic biotinable minimal peptide, competitively inhibits self-biotinylation
glycerol
-
inhibits the recombinant enzyme at 10%
MgAMP
-
endproduct inhibition, mechanism of inhibition, overview
NaCl
-
inhibits the purified enzyme at 0.1 M, but not the immobilized enzyme in crude extract of recombinant yeast cells
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0052
apo-[acetyl-CoA:carbon-dioxide ligase (ADP-forming)]
-
pH 8.0, 37°C
0.018 - 0.42
ATP
0.0033
avitag-tagged maltose-binding protein
-
recombinant enzyme
-
0.000026 - 0.26
biotin
0.000424
d-biotin
-
pH 8.0, 37°C
0.2 - 5.1
MgATP2-
0.13
yeast acceptor peptide
-
at 25°C
-
0.12
[Escherichia coli apo-biotin-carboxyl-carrier-protein mutant M136S]-L-lysine
-
pH 8.0, 37°C, wild type enzyme
-
0.011
[Escherichia coli apo-biotin-carboxyl-carrier-protein]-L-lysine
-
pH 8.0, 37°C, wild type enzyme
-
0.03 - 0.101
[Sulfolobus tokodaii apo-biotin-carboxyl-carrier-protein mutant S136M]-lysine
-
0.00044 - 0.00058
[Sulfolobus tokodaii apo-biotin-carboxyl-carrier-protein]-lysine
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
440 - 360000
ATP
0.037 - 450000
biotin
0.081 - 0.32
MgATP2-
0.0017
yeast acceptor peptide
Saccharomyces cerevisiae
-
at 25°C
-
0.58
[Escherichia coli apo-biotin-carboxyl-carrier-protein mutant M136S]-L-lysine
Escherichia coli
-
pH 8.0, 37°C, wild type enzyme
-
0.165
[Escherichia coli apo-biotin-carboxyl-carrier-protein]-L-lysine
Escherichia coli
-
pH 8.0, 37°C, wild type enzyme
-
0.0097 - 0.165
[Sulfolobus tokodaii apo-biotin-carboxyl-carrier-protein mutant S136M]-lysine
-
0.0022 - 0.004
[Sulfolobus tokodaii apo-biotin-carboxyl-carrier-protein]-lysine
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.49
[Escherichia coli apo-biotin-carboxyl-carrier-protein mutant M136S]-L-lysine
Escherichia coli
-
pH 8.0, 37°C, wild type enzyme
42694
15
[Escherichia coli apo-biotin-carboxyl-carrier-protein]-L-lysine
Escherichia coli
-
pH 8.0, 37°C, wild type enzyme
42693
0.32 - 1.6
[Sulfolobus tokodaii apo-biotin-carboxyl-carrier-protein mutant S136M]-lysine
12261
3.8 - 9.1
[Sulfolobus tokodaii apo-biotin-carboxyl-carrier-protein]-lysine
10432
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.01
(3aS,4S,6aR)-4-(5-[1-[3-(naphthalen-2-yloxy)propyl]-1H-1,2,3-triazol-4-yl]pentyl)tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one
-
Ki above 0.01 mM, wild type enzyme, at pH 7.5 and 37°C
0.01
(3aS,4S,6aR)-4-(5-[1-[4-(naphthalen-1-yloxy)butyl]-1H-1,2,3-triazol-4-yl]pentyl)tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one
-
Ki above 0.01 mM, wild type enzyme, at pH 7.5 and 37°C
0.0017
(3aS,4S,6aR)-4-(5-[1-[4-(naphthalen-2-yloxy)butyl]-1H-1,2,3-triazol-4-yl]pentyl)tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one
-
wild type enzyme, at pH 7.5 and 37°C
0.00117
(3aS,4S,6aR)-4-[-[1-[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl]methyl]triazol-4-yl]pentyl]-1,3,3a,4,6,6ahexahydrothieno[3,4-d]imidazol-2-one
-
wild type enzyme, at pH 7.5 and 37°C
0.01
(3aS,4S,6aR)-4-[5-[1-(4-phenoxybutyl)-1H-1,2,3-triazol-4-yl]pentyl]tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one
-
Ki above 0.01 mM, wild type enzyme, at pH 7.5 and 37°C
0.01
2-(4-[5-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentyl]-1H-1,2,3-triazol-1-yl)ethyl 2,2-dimethylpropanoate
-
Ki above 0.01 mM, wild type enzyme, at pH 7.5 and 37°C
0.01
5-methyl-3-[3-(4-[5-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentyl]-1H-1,2,3-triazol-1-yl)propyl]-1,3-benzoxazol-2(3H)-one
-
Ki above 0.01 mM, wild type enzyme, at pH 7.5 and 37°C
0.00009
5-methyl-3-[4-(4-[5-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentyl]-1H-1,2,3-triazol-1-yl)butyl]-1,3-benzoxazol-2(3H)-one
-
wild type enzyme, at pH 7.5 and 37°C
0.36 - 0.62
ATP
0.0003 - 0.0076
biotin acetylene
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
8.24
MgAMP
Escherichia coli
-
pH 8.0, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000046
-
-
0.049
-
purified recombinant enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.3
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36
-
wild type enzyme and mutant enzymes birA311, birA825, birA301, and birA352
65
binding assays at
80
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
lung fibroblasts, quantitative determination of HCS in wild-type IMR-90 fibroblasts and in those recombinantly overexpressing human telomerase, evaluation as a cell model for investigation of HCS in epigenetic regulation, histone biotinylation in aging cells, overview
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
the longer Met58 isoform
Manually annotated by BRENDA team
additional information
-
isozyme subcellular localization, immunofluorescence study, overview
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain CDC 1551 / Oshkosh)
Mycobacterium tuberculosis (strain CDC 1551 / Oshkosh)
Mycobacterium tuberculosis (strain CDC 1551 / Oshkosh)
Mycobacterium tuberculosis (strain CDC 1551 / Oshkosh)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7909
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liquid-chromatography electrospray ionisation ion-trap mass spectroscopy
26000
2 * 26000, calculated. Equilibrium analytical ultracentrifugation measurements performed on the apoenzyme and its complexes with biotin and bio-5-AMP all yield molecular weights for the protein consistent with a dimer. Regardless of ligation state the Pyrococcus horikoshii enzyme is a dimer
26071
-
2 * 26071 recombinant enzyme, SDS-PAGE and amino acid sequence calculation
26630
-
MALDI TOF mass spectrometry
29500
-
monomeric enzyme, gel filtration
35000
-
x * 35000, recombinant enzyme, SDS-PAGE
35300
-
SDS-PAGE
35310
-
MALDI TOF mass spectrometry
38300
-
2 * 38300, calculated from amino acid sequence
51200
sedimentation equilibrium study
53900
-
recombinant enzyme, light scattering study
56000
-
dimeric enzyme, gel filtration
73000
-
x * 73000, recombinant His9-tagged BPL, SDS-PAGE
76000
-
sedimentation equilibrium analysis
82000
-
SDS-PAGE
265000
-
acetyl-CoA carboxylase-1
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant wild-type and mutant R40G enzymes free and in complex with ligands ATP and/or biotin, hanging drop vapour diffusion method, 0.002 ml of protein solution with 5 or 6 mg/ml wild-type or mutant protein, respectively, is mixed with 0.001 ml of reservoir solution containing 0.1M Mes, 0.2 M ammonium sulfate, 15% w/v PEG 5000 monoethyl ether, pH 6.5, with or without 2 mM D-biotin and 5 mM ATP, 17°C, a few hours for the free enzyme, 4-14 days for the complexed enzyme, X-ray diffraction structure determination and analysis at 2.3-2.55 A resolution, molecular modeling
EcBPL domains determination through X-ray crystallography at 2.3 A resolution, and determination of the crystal structure of EcBPL in complex with biotin
purified recombinant enzyme, 0.003 ml of 5 mg/ml protein in 2.5 mM desthiobiotin in 20 mM Tris-HCl pH 8.0, 50 mM NaCl, 1 mM PMSF and 1 mM DTT, is mixed with 0.003 ml of reservoir solution containing 12-16% w/v of both PEG 4000 and PEG 8000 in 0.1 M HEPES, pH 7.5, X-ray diffraction structure determination and analysis at 2.8 A resolution
structures of dehydrated and hydrated BirA, at 2.69 A and 2.8 A resolution, respectively. Dehydration of BirA crystals traps both the apo and active conformations in its asymmetric unit. The crystal lattice rearrangement due to shrinkage in the dehydrated Mtb-BirA crystals ensues structural order of otherwise flexible ligand-binding loops L4 and L8 in apo BirA. In addition, crystal dehydration results in a shift of 3.5 A in the flexible loop L6, a proline-rich loop unique to Mycobacterium tuberculosis complex as well as around the L11 region. The shift in loop L11 in the C-terminal domain on dehydration emulates the action responsible for the complex formation with its protein ligand biotin carboxyl carrier protein domain of ACCA3. The two subunits A and B, though related by a noncrystallographic twofold symmetry, assemble into an asymmetric dimer representing the ligand-bound and ligand-free states of the protein, respectively
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oil-microbatch method using PEG 20000 as a precipitant
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PhBPL in complex with biotin, ATP and biotinyl-5'-AMP, X-ray diffraction and structure analysis at 1.6 A, 1.6 A, 1.45 A resolution, respectively. Structure-function relationship, modelling, overview
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purified recombinant enzyme, complexed with biotinyl-5'-AMP, biotin, adenosine, or mutant biotin carboxyl carrier protein PhBCCPN76, X-ray diffraction structure determination at 2.0-2.7 A resolution, analysis, and modelling. The non-hydrolyzable intermediate analogue biotinol-5'-AMP does not provide analyzable complex crystals
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purified recombinant native and selenomethionine-labeled enzyme comprising residues 1-125, sitting drop method, 9.87 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 500 mM NaCl, 0.001 ml protein solution is mixed with 0.001 ml precipitation solution containing 10.5% PEG 2000, 0.1 M acetate-NaOH, pH 5.5, the drop is overlaid with a 1:1 mixture of silicone and paraffin oil allowing slow water evaporation from the drop, 22°C, preliminary X-ray diffraction structure determination and analysis at 1.45-1.6 A resolution
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purified recombinant native and selenomethionine-labeled enzyme, sitting drop method, 9.87 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 500 mM NaCl, 0.001 ml protein solution is mixed with 0.001 ml precipitation solution containing 10.5% PEG 2000, 0.1 M acetate-NaOH, pH 5.5, the drop is overlaid with a 1:1 mixture of silicone and paraffin oil allowing slow water evaporation from the drop, 22°C, preliminary X-ray diffraction structure determination and analysis at 1.45-1.6 A resolution
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apoenzyme, in complex with biotin, and complexed with biotinyl-5'-AMP, hanging drop vapor diffusion method, using
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hanging drop vapor diffusion method, using 8-12% (w/v) PEG8000 in 0.1 M Tris pH 7.5 or 8.0 and 10% (v/v) glycerol
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purified recombinant enzyme, hanging-drop vapour-diffusion method, 0.001 ml of 15 mg/ml protein solution is mixed with 0.001 ml of reservoir solution containing 8% PEG 8000 and 0.1 M Tris–HCl pH 8.0., at 4°C or 20°C, crystals appear within 24 h, X-ray diffraction structure determination and analysis at 2.3 A resolution
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
80
-
no denaturation
additional information
-
mutant enzymes birA707, birA361 and birA91 with strikingly altered thermosensitivity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, 1-2 mg/ml purified recombinant His6-tagged enzyme, in 50 mM Tris-HCl, pH 7.5, 0.5 mM EDTA, pH 8.0, 250 mM NaCl, at least 6 months without loss of activity
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4°C, immobilized recombinant enzyme, PBS, completely stable for 4 weeks, loss of activity after 5 months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
HisTrap column chromatography, Ni-NTA resin column chromatography, and Superdex 75 gel filtration
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mutants K277E, R317E, and K321E
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Ni-NTA column chromatography, and Superdex 200 gel filtration
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recombinant C-terminally Strep-tagged BirA from Escherichia coli
recombinant enzyme comprising residues 1-125 from Escherichia coli strain BL21
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recombinant enzyme from Escherichia coli
recombinant enzyme from Escherichia coli strain BL21 in a chromatographic procedure comprising several steps
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recombinant enzyme from Escherichia coli, the human enzyme degrades significantly during isolation
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recombinant enzyme from Saccharomyces cerevisiae, immobilization of the recombinant enzyme
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recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration
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recombinant His-tagged wild-type and mutant enzymes from strain JM109 by nickel affinity chromatography
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recombinant His6-tagged enzyme from strain BL21(DE3) by nickel affinity chromatography
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recombinant His9-tagged BPL from Saccharomyces cerevisiae strain w303 by nickel affinity and cation exchange chromatography
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recombinant human HCS from Hi5 insect cells by nickel affinity chromatography, anion exchange chromatography, and gel filtration
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recombinant protein
Super Q Toyopearl 650M column chromatography, Bio-Scale CHT-20-I column chromatography, and HiLoad 16/60 Superdex 200 gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
coding region of birA biotin-protein ligase gene cloned from genomic DNA by PCR, verfied by sequencing and expressed in mouse erythroleukemic cells
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construction and expression of the C-terminal 87 amino acids of biotin carboxyl carrier protein, i.e. BCCP-87. cross-species activity of BPLs permitts functional cloning of the cDNA for yeast BPL through genetic complementation with a conditional lethal birA- strain of Escherichia coli
cross-species activity of BPLs permitts functional cloning of the cDNA for human BPL through genetic complementation with a conditional lethal birA? strain of Escherichia coli
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cross-species activity of BPLs permitts functional cloning of the cDNA for yeast BPL through genetic complementation with a conditional lethal birA? strain of Escherichia coli
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DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli strain BL21
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doxycycline-regulated expression of a secreted variant of Escherichia coli biotin ligase BirA with target proteins containing a 13-residue biotin acceptor peptide appended to their extracellular domain, co-expression with three different G-protein coupled receptors, i.e. protease-activated receptors 1 and 2, and the platelet ADP receptor, P2Y12, which become biotinylated, in CHO-K1 Tet-On cells on the cell surface, overview
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expressed as thioredoxin- and maltose binding protein-fusion proteins in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli BL21 cells
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expressed in Escherichia coli DH5alpha cells
expression as His6-tagged enzyme in strain BL21(DE3)
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expression in Escherichia coli
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expression in Escherichia coli BL21(DE3)
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expression in Escherichia coli strain JM109
expression in Escherichia coli strains DH5alpha and BL21(DE3)
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expression of BirA from pET-based vector pHBA in Escherichia coli BL21 (lambdaDE3)
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expression of HCS in Spodoptera frugiperda Sf9 cells and in Hi5 insect cells using the baculovirus transfection system
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expression of His9-tagged BPL in Saccharomyces cerevisiae strain w303, functional complementation of the conditional lethal birA85 Escherichia coli strain BM4062
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gene birA, expression of His-tagged wild-type and mutant enzymes in strain JM109, expression of enzyme in strain BM4092 unbiotinylated form
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gene birA, functional expression of N-terminally and C-terminally multiply tagged enzyme in Saccharomyces cerevisiae, low level expression of the surface-displayed enzyme, expression of avitagged-BirA in Saccharomyces cerevisiae, overview
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gene Rv3279c, DNA and amino acid sequence determination and analysis, expression of C-terminally Strep-tagged BirA in Escherichia coli
overexpression in Escherichia coli
overexpression of the enzyme comprising residues 1-125 in Escherichia coli strain BL21
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
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
I272X
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site-directed mutagenesis
K122E
-
site-directed mutagenesis
K277E
-
site-directed mutagenesis
K283E
-
site-directed mutagenesis
K321E
-
site-directed mutagenesis
L237P
the mutant shows reduced catalytic activity compared to the wild-type enzyme
R182E
-
site-directed mutagenesis
R212E
-
site-directed mutagenesis
R213E
-
site-directed mutagenesis
R317E
-
site-directed mutagenesis
D571N
-
naturally occuring mutation, important in positioning K579 in the AMP binding site
D615Y
-
naturally occuring mutation in the loop between alpha3 and beta6 that cover AMP, may coordinate oxygens of the AMP phosphate
D634Y
-
naturally occuring mutation of a solvent exposed residue, distal to active site on alpha4 and alpha5, respectively
D715G
-
naturally occuring mutation on beta9, may be involved in capping and stabilising the catalytic domain structure
G518E
-
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
-
naturally occuring mutation of a residue involved in hydrophobic interactions with biotin in the binding pocket
L470S
-
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
-
naturally occuring mutation in an unstructured loop distal to the ligand-binding site
V547G
-
naturally occuring mutation of a buried hydrophobic residue in alpha2 that reside near the AMP-binding beta5 strand
V550M
-
naturally occuring mutation of a buried hydrophobic residue in alpha2 that reside near the AMP-binding beta5 strand
Y456C
-
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
-
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
-
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
-
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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F123G
-
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
-
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
-
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
diagnostics
-
BirA can be useful for specific in vivo labeling of proteins in cell cultures by biotinylation
drug development
synthesis
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