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Literature summary for 6.3.4.6 extracted from

  • Fan, C.; Chou, C.Y.; Tong, L.; Xiang, S.
    Crystal structure of urea carboxylase provides insights into the carboxyltransfer reaction (2012), J. Biol. Chem., 287, 9389-9398.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.3.4.6

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Kluyveromyces lactis

Crystallization (Commentary)

Crystallization (Comment) Organism
sitting drop vapor diffusion method, using 2.4 M ammonium sulfate and 0.1 M bis-Tris, pH 7.0 Kluyveromyces lactis

Protein Variants

Protein Variants Comment Organism
D1321A the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Kluyveromyces lactis
D1584N the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Kluyveromyces lactis
E1792A the mutant shows reduced catalytic efficiency compared to the wild type enzyme Kluyveromyces lactis
K1605A the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Kluyveromyces lactis
Y1324F the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Kluyveromyces lactis
Y1628F the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Kluyveromyces lactis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.32
-
 Urea wild type enzyme, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
0.39
-
 Urea mutant enzyme E1792A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
1 1.3 Urea mutant enzyme D1564N, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
7.7
-
 Urea mutant enzyme Y1324F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
15
-
 Urea mutant enzyme Y1628F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
78
-
 Urea mutant enzyme D1321A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
493
-
 Urea mutant enzyme K1605A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
137000
-
 1 * 137000, calculated from amino acid sequence Kluyveromyces lactis
141000
-
 gel filtration Kluyveromyces lactis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + urea + HCO3- + H+ Kluyveromyces lactis
-
 ADP + phosphate + urea-1-carboxylate
-
 ?
ATP + urea + HCO3- + H+ Kluyveromyces lactis ATCC 8585
-
 ADP + phosphate + urea-1-carboxylate
-
 ?

Organism

Organism UniProt Comment Textmining
Kluyveromyces lactis Q6CP22
-
-
Kluyveromyces lactis ATCC 8585 Q6CP22
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA agarose column chromatography and Sephacryl S-300 gel filtration Kluyveromyces lactis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + urea + HCO3- + H+
-
 Kluyveromyces lactis ADP + phosphate + urea-1-carboxylate
-
 ?
ATP + urea + HCO3- + H+
-
 Kluyveromyces lactis ATCC 8585 ADP + phosphate + urea-1-carboxylate
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 137000, calculated from amino acid sequence Kluyveromyces lactis
monomer 1 * 141000, dynamic light scattering Kluyveromyces lactis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.44
-
 Urea mutant enzyme K1605A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
3.16
-
 Urea mutant enzyme D1321A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
45.8
-
 Urea mutant enzyme Y1324F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
61.8
-
 Urea mutant enzyme D1564N, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
64.9
-
 Urea mutant enzyme Y1628F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
68.7
-
 Urea mutant enzyme E1792A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
210.8
-
 Urea wild type enzyme, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Kluyveromyces lactis
biotin
-
 Kluyveromyces lactis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0029
-
 Urea mutant enzyme K1605A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
0.041
-
 Urea mutant enzyme D1321A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
4.3
-
 Urea mutant enzyme Y1628F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
5.5
-
 Urea mutant enzyme D1564N, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
5.9
-
 Urea mutant enzyme Y1324F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
176
-
 Urea mutant enzyme E1792A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis
659
-
 Urea wild type enzyme, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C Kluyveromyces lactis