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Literature summary for 6.3.4.14 extracted from

  • Chou, C.Y.; Tong, L.
    Structural and biochemical studies on the regulation of biotin carboxylase by substrate inhibition and dimerization (2011), J. Biol. Chem., 286, 24417-24425.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

Crystallization (Comment) Organism
sitting drop vapor diffusion method, using 17% (w/v) PEG3350 and 0.14 M CaCl2, (in complex with two ADP and two Ca2+ ions), or 0.1 M Bis-tris (pH 6.2), 15% (w/v) PEG3350, 0.1 M NH4Cl, and 5% (w/v) n-octyl-beta-D-glucoside (in complex with two ADP and one Mg2+ ion), or 18% (w/v) PEG3350, 0.17 M CsCl, and 4% (v/v) methanol (R16E mutant in complex with bicarbonate and Mg2+-ADP) Escherichia coli

Protein Variants

Protein Variants Comment Organism
R16E the mutant has a 2fold loss in catalytic activity compared with the wild type enzyme. The mutation significantly destabilizes the dimer Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
ADP
-
Escherichia coli
ATP ATP shows substrate inhibition which is competitive against bicarbonate Escherichia coli
CTP less than 40% residual activity at 20 mM Escherichia coli
GTP less than 40% residual activity at 20 mM Escherichia coli
TTP less than 40% residual activity at 20 mM Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1 2 ATP mutant enzyme R16E, in 100 mM HEPES (pH 8.0), 5 mM MgCl2, at 30°C Escherichia coli
0.1 2 ATP wild type enzyme, in 100 mM HEPES (pH 8.0), 5 mM MgCl2, at 30°C Escherichia coli
1.9
-
HCO3- wild type enzyme, with 1.5 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli
5
-
HCO3- wild type enzyme, with 7.5 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli
6.5
-
HCO3- wild type enzyme, with 10 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli
11.4
-
HCO3- wild type enzyme, with 20 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ contains Ca2+ Escherichia coli
Mg2+ dependent on Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + biotin-carboxyl-carrier protein + HCO3- Escherichia coli
-
ADP + phosphate + carboxybiotin-carboxyl-carrier protein
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli P24182
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + biotin-carboxyl-carrier protein + HCO3-
-
Escherichia coli ADP + phosphate + carboxybiotin-carboxyl-carrier protein
-
?

Subunits

Subunits Comment Organism
dimer
-
Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.2
-
ATP mutant enzyme R16E, in 100 mM HEPES (pH 8.0), 5 mM MgCl2, at 30°C Escherichia coli
0.3
-
HCO3- wild type enzyme, with 20 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli
0.31
-
HCO3- wild type enzyme, with 1.5 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli
0.32
-
HCO3- wild type enzyme, with 10 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli
0.36
-
ATP wild type enzyme, in 100 mM HEPES (pH 8.0), 5 mM MgCl2, at 30°C Escherichia coli
0.36
-
HCO3- wild type enzyme, with 7.5 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
Escherichia coli

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
27
-
ATP wild type enzyme, in 100 mM HEPES (pH 8.0), 5 mM MgCl2, at 30°C Escherichia coli

General Information

General Information Comment Organism
metabolism the biotin carboxylase subunit of acetyl-CoA carboxylase catalyzes the Mg2+-ATP-dependent carboxylation of biotin Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
26
-
HCO3- wild type enzyme, with 20 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli
49
-
HCO3- wild type enzyme, with 10 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli
71
-
HCO3- wild type enzyme, with 7.5 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli
162
-
HCO3- wild type enzyme, with 1.5 mM Mg-ATP, in 100 mM HEPES (pH 8.0), at 30°C Escherichia coli
1667
-
ATP mutant enzyme R16E, in 100 mM HEPES (pH 8.0), 5 mM MgCl2, at 30°C Escherichia coli
3000
-
ATP wild type enzyme, in 100 mM HEPES (pH 8.0), 5 mM MgCl2, at 30°C Escherichia coli