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Literature summary extracted from

  • Chou, C.Y.; Yu, L.P.; Tong, L.
    Crystal structure of biotin carboxylase in complex with substrates and implications for its catalytic mechanism (2009), J. Biol. Chem., 284, 11690-11697.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
6.3.4.14 wild-type Escherichia coli biotin carboxylase and mutant E296A in complex with its substrates biotin, bicarbonate, and Mg-ADP, at 2.0 A resolution. Residue Glu296 is the general base that extracts the proton from bicarbonate, and Arg338 is the residue that stabilizes the enolate biotin intermediate in the carboxylation reaction. The B domain of biotin carboxylase is positioned closer to the active site, leading to a 2-A shift in the bound position of the adenine nucleotide and bringing it near the bicarbonate for catalysis. One of the oxygenatoms of bicarbonate is located in the correct position to initiate the nucleophilic attack on ATP to form the carboxyphosphate intermediate. The phosphate group, derived from decomposition of carboxyphosphate, is the general base that extracts the proton on this N1 atom Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
6.3.4.14 E296A 50fold decrease in catalytic efficiency, crystallization data Escherichia coli
6.3.4.14 R338A 250fold decrease in catalytic efficiency Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.4.14 16.2
-
 HCO3- pH 8.5, wild-type Escherichia coli
6.3.4.14 18.9
-
 HCO3- pH 8.5, mutant R338A Escherichia coli
6.3.4.14 22.7
-
 biotin pH 7.4, mutant E296A Escherichia coli
6.3.4.14 35.1
-
 biotin pH 7.4 wild-type Escherichia coli
6.3.4.14 41.8
-
 biotin pH 7.4, mutant R338A Escherichia coli
6.3.4.14 57.5
-
 HCO3- pH 8.5, mutant E296A Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
6.3.4.14 Escherichia coli P24182
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.4.14 ATP + biotin + HCO3-
-
 Escherichia coli ADP + phosphate + carboxybiotin
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.3.4.14 0.0019
-
 HCO3- pH 8.5, mutant R338A Escherichia coli
6.3.4.14 0.0025
-
 biotin pH 7.4, mutant R338A Escherichia coli
6.3.4.14 0.028
-
 biotin pH 7.4, mutant E296A Escherichia coli
6.3.4.14 0.034
-
 HCO3- pH 8.5, mutant E296A Escherichia coli
6.3.4.14 0.44
-
 HCO3- pH 8.5, wild-type Escherichia coli
6.3.4.14 0.58
-
 biotin pH 7.4 wild-type Escherichia coli

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.3.4.14 0.00006
-
 biotin pH 7.4, mutant R338A Escherichia coli
6.3.4.14 0.0001
-
 HCO3- pH 8.5, mutant R338A Escherichia coli
6.3.4.14 0.000594
-
 HCO3- pH 8.5, mutant E296A Escherichia coli
6.3.4.14 0.00123
-
 biotin pH 7.4, mutant E296A Escherichia coli
6.3.4.14 0.0165
-
 biotin pH 7.4 wild-type Escherichia coli
6.3.4.14 0.0272
-
 HCO3- pH 8.5, wild-type Escherichia coli