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Literature summary for 3.5.4.1 extracted from

  • Hall, R.S.; Fedorov, A.A.; Xu, C.; Fedorov, E.V.; Almo, S.C.; Raushel, F.M.
    Three-dimensional structure and catalytic mechanism of cytosine deaminase (2011), Biochemistry, 50, 5077-5085.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.5.4.1

Crystallization (Commentary)

Crystallization (Comment) Organism
the structure of Zn-CDA is determined to a resolution of 1.7 A with phosphonocytosine a potent mimic of the putative tetrahedral intermediate bound in the active site Escherichia coli

Protein Variants

Protein Variants Comment Organism
D313A metal contet: 0.57 Zn, 0.32 Fe. Mutant possesses less than 0.01% of activity of wild-type enzyme Escherichia coli
D313N metal contet: 1.05 Zn, 0.05 Fe. kcat decreased compared to wild-type, Km increased compared to wild-type Escherichia coli
E217A metal contet: 0.9 Zn, 0.08 Fe. Mutant possesses less than 0.01% of activity of wild-type enzyme Escherichia coli
E217Q metal contet: 0.87 Zn, 0.15 Fe. Mutant possesses less than 0.01% of activity of wild-type enzyme Escherichia coli
H246A metal contet: 0.59 Zn, 0.33 Fe. Mutant possesses less than 0.01% of activity of wild-type enzyme Escherichia coli
H246N metal contet: 0.53 Zn, 0.29 Fe. Mutant possesses less than 0.01% of activity of wild-type enzyme Escherichia coli
H246Q metal contet: 0.31 Zn, 0.33 Fe. kcat and Km decreased compared to wil-type Escherichia coli
Q156A metal contet: 0.92 Zn, 0.04 Fe. Mutant possesses less than 0.01% of activity of wild-type enzyme Escherichia coli
Q156N metal contet: 1.40 Zn, 0.04 Fe. Mutant possesses less than 0.01% of activity of wild-type enzyme Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
phosphonocytosine
-
 Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.25
-
 cytosine mutant H246Q, pH 8.5, 30°C Escherichia coli
0.46
-
 isocytosine wild-type, pH 8.5, 30°C Escherichia coli
0.85
-
 3-oxauracil mutant E217A, pH 8.5, 30°C Escherichia coli
0.97
-
 cytosine wild-type, pH 8.5, 30°C Escherichia coli
1.9
-
 cytosine mutant D313N, pH 8.5, 30°C Escherichia coli
4.1
-
 3-oxauracil wild-type, pH 8.5, 30°C Escherichia coli
25
-
 creatinine wild-type, pH 8.5, 30°C Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Iron the enzyme contains 0.20 equivalents of Fe Escherichia coli
Zinc the enzyme contained 0.56 equivalents of Zn Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-oxauracil + H2O
-
 Escherichia coli malonate semialdehyde + CO2 + NH3
-
 ?
creatinine + H2O
-
 Escherichia coli N-methylhydantoin + NH3
-
 ?
cytosine + H2O
-
 Escherichia coli uracil + NH3
-
 ?
isocytosine + H2O
-
 Escherichia coli ?
-
 ?

Synonyms

Synonyms Comment Organism
CDA
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0075
-
 cytosine mutant D313N, pH 8.5, 30°C Escherichia coli
0.18
-
 3-oxauracil mutant E217A, pH 8.5, 30°C Escherichia coli
0.18
-
 cytosine mutant H246Q, pH 8.5, 30°C Escherichia coli
2.3
-
 3-oxauracil wild-type, pH 8.5, 30°C Escherichia coli
5.1
-
 isocytosine wild-type, pH 8.5, 30°C Escherichia coli
5.6
-
 creatinine wild-type, pH 8.5, 30°C Escherichia coli
132
-
 cytosine wild-type, pH 8.5, 30°C Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
 assay at Escherichia coli

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000052
-
 phosphonocytosine pH 8.5, 30°C Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.004
-
 cytosine mutant D313N, pH 8.5, 30°C Escherichia coli
0.21
-
 3-oxauracil mutant E217A, pH 8.5, 30°C Escherichia coli
0.22
-
 creatinine wild-type, pH 8.5, 30°C Escherichia coli
0.56
-
 3-oxauracil wild-type, pH 8.5, 30°C Escherichia coli
0.72
-
 cytosine mutant H246Q, pH 8.5, 30°C Escherichia coli
11
-
 isocytosine wild-type, pH 8.5, 30°C Escherichia coli
140
-
 cytosine wild-type, pH 8.5, 30°C Escherichia coli