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Literature summary extracted from

  • Mochalkin, I.; Miller, J.R.; Evdokimov, A.; Lightle, S.; Yan, C.; Stover, C.K.; Waldrop, G.L.
    Structural evidence for substrate-induced synergism and half-sites reactivity in biotin carboxylase (2008), Protein Sci., 17, 1706-1718.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
6.3.4.14 biotin substrate induced synergism, the reaction inpresence of biotin is 1100fold faster than in its absence Pseudomonas aeruginosa
6.3.4.14 biotin substrate-induced synergism, the reaction in presence of biotin is 1100fold faster than in its absence Staphylococcus aureus
6.3.4.14 biotin substrate-induced synergism, the reaction in presence of biotin is 1100fold faster than in its absence Escherichia coli

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.4.14 gene accC, expression in Escherichia coli srrain BL21-AI as His-tagged enzyme Pseudomonas aeruginosa
6.3.4.14 gene accC, expression in Escherichia coli strain BL21-AI as His-tagged enzyme Escherichia coli
6.3.4.14 gene accC, overexpression in Escherichia coli strain BL21(DE3) as His-tagged enzyme Staphylococcus aureus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
6.3.4.14 enzyme in complex with ATP analogues AMP-PNP and ADP-CF2P, hanging drop vapour diffusion, from 0.1 M Bis-Tris, pH 6.5, 0.2 M CaCl2, 45% methylpentanediol, and 10% ethylene glycol or 0.1 M KCl, 3-8% PEG 8000 and 20% ethylene glycol, respectively, X-ray diffraction structure determination and analysis at 2.05 and 2.69 A resolution, respectively, structure modelling Escherichia coli
6.3.4.14 recombinant apoenzyme or enzyme in complex with ATP analogue AMP-PCP, sitting drop vapour diffusion, from 0.1 M HEPES, pH 7.0, 0.2 M MgCl2, 15-20% PEG 3350, and 10% ethylene glycol in the well solution, X-ray diffraction structure determination and analysis at 2.4 A resolution, structure modelling Pseudomonas aeruginosa
6.3.4.14 recombinant enzyme in complex with ATP analogue AMP-PNP, sitting drop vapour diffusion, from 0.2 M KCl, 20% PEG 3350, and 20% ethylene glycol in the well solution, X-ray diffraction structure determination and analysis at 2.10, structure modelling Staphylococcus aureus

Protein Variants

EC Number Protein Variants Comment Organism
6.3.4.14 G165V site-directed mutagenesis, the active site mutant has the residue of the Staphylococcus aureus enzyme and shows increased Km for ATP and 100fold decreased reaction velocity compared to the wild-type enzyme Escherichia coli
6.3.4.14 G166V site-directed mutagenesis, the active site mutant has the residue of the Staphylococcus aureus enzyme and shows increased Km for ATP and 100fold decreased reaction velocity compared to the wild-type enzyme Escherichia coli
6.3.4.14 K116Q site-directed mutagenesis, the phosphate binding site mutant has the residue of the Staphylococcus aureus enzyme and shows a 50fold increased Km for ATP compared to the wild-type enzyme Escherichia coli
6.3.4.14 K159Q site-directed mutagenesis, the active site mutant has the residue of the Staphylococcus aureus enzyme and shows a 90fold higher Km for ATP compared to the wild-type enzyme Escherichia coli
6.3.4.14 R292A site-directed mutagenesis, the mutant has a Km for ATP similar to the wild-type enzyme Escherichia coli

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.4.14 Mg2+ binding sites structure Staphylococcus aureus
6.3.4.14 Mg2+ binding sites structure Escherichia coli
6.3.4.14 Mg2+ binding sites structure Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.4.14 ATP + biotin-carboxyl-carrier protein + CO2 Staphylococcus aureus
-
 ADP + phosphate + carboxybiotin-carboxyl-carrier protein
-
 ?
6.3.4.14 ATP + biotin-carboxyl-carrier protein + CO2 Escherichia coli
-
 ADP + phosphate + carboxybiotin-carboxyl-carrier protein
-
 ?
6.3.4.14 ATP + biotin-carboxyl-carrier protein + CO2 Pseudomonas aeruginosa
-
 ADP + phosphate + carboxybiotin-carboxyl-carrier protein
-
 ?
6.3.4.14 ATP + biotin-carboxyl-carrier protein + CO2 Staphylococcus aureus Mu50
-
 ADP + phosphate + carboxybiotin-carboxyl-carrier protein
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.4.14 Escherichia coli
-
 gene accC
-
6.3.4.14 Pseudomonas aeruginosa
-
-
-
6.3.4.14 Staphylococcus aureus
-
-
-
6.3.4.14 Staphylococcus aureus Mu50
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.4.14 recombinant His-tagged enzyme from Escherichia coli srrain BL21-AI by nickel affinity and anion exchange chromatography, and gel filtration Pseudomonas aeruginosa
6.3.4.14 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Staphylococcus aureus
6.3.4.14 recombinant His-tagged enzyme from Escherichia coli strain BL21-AI by nickel affinity chromatography and gel filtration Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.4.14 ATP + biotin-carboxyl-carrier protein + CO2
-
 Staphylococcus aureus ADP + phosphate + carboxybiotin-carboxyl-carrier protein
-
 ?
6.3.4.14 ATP + biotin-carboxyl-carrier protein + CO2
-
 Escherichia coli ADP + phosphate + carboxybiotin-carboxyl-carrier protein
-
 ?
6.3.4.14 ATP + biotin-carboxyl-carrier protein + CO2
-
 Pseudomonas aeruginosa ADP + phosphate + carboxybiotin-carboxyl-carrier protein
-
 ?
6.3.4.14 ATP + biotin-carboxyl-carrier protein + CO2
-
 Staphylococcus aureus Mu50 ADP + phosphate + carboxybiotin-carboxyl-carrier protein
-
 ?
6.3.4.14 additional information active site structure, substrate binding and structure-function relationship analysis, overview Staphylococcus aureus ?
-
 ?
6.3.4.14 additional information active site structure, substrate binding and structure-function relationship analysis, overview Escherichia coli ?
-
 ?
6.3.4.14 additional information active site structure, substrate binding and structure-function relationship analysis, overview Pseudomonas aeruginosa ?
-
 ?
6.3.4.14 additional information active site structure, substrate binding and structure-function relationship analysis, overview Staphylococcus aureus Mu50 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
6.3.4.14 dimer homodimer Staphylococcus aureus
6.3.4.14 dimer homodimer Escherichia coli
6.3.4.14 dimer homodimer Pseudomonas aeruginosa

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.4.14 ATP
-
 Staphylococcus aureus
6.3.4.14 ATP
-
 Escherichia coli
6.3.4.14 ATP
-
 Pseudomonas aeruginosa