Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.5.4.5 extracted from

  • Johansson, E.; Neuhard, J.; Willemoes, M.; Larsen, S.
    Structural, kinetic, and mutational studies of the zinc ion environment in tetrameric cytidine deaminase (2004), Biochemistry, 43, 6020-6029.
    View publication on PubMed
Search for more literature for 3.5.4.5

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of wild-type enzyme and mutant enzymes R56A, R56Q and C53H/R56Q in Escherichia coli Bacillus subtilis

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapour diffusion method. R56A and R56Q crystallize in the same space group as the wild-type enzyme with two subunits in the asymmetric unit, whereas C53H/R56Q can not crystallize in this crystal form but is crystallized in another space group, P3(2)21, with a full tetramer in the asymmetric unit Bacillus subtilis

Protein Variants

Protein Variants Comment Organism
C53H attempts to express and purify the mutant enzyme are unsuccessful Bacillus subtilis
C53H/R56Q mutant enzyme contains the same amount of zinc as the wild-type enzyme. Because of dissociation into its inactive subunits, it is impossible to determine the kinetic parameters for the mutant enzyme Bacillus subtilis
R56A mutant enzyme contains the same amount of zinc as the wild-type enzyme. Similar KM-value but decreased Vmax-value compared to wild-type enzyme Bacillus subtilis
R56D zinc-binding capacity of mutant enzyme is reduced Bacillus subtilis
R56Q mutant enzyme contains the same amount of zinc as the wild-type enzyme. Similar KM-value but decreased Vmax-value compared to wild-type enzyme Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0075
-
 deoxycytidine pH 7.8, 25°C, mutant enzyme R56A Bacillus subtilis
0.029
-
 deoxycytidine pH 7.8, 25°C, mutant enzyme R56Q Bacillus subtilis
0.14
-
 cytidine pH 7.6, 25°C, mutant enzyme R56A Bacillus subtilis
0.216
-
 cytidine pH 7.6, 25°C, wild-type enzyme Bacillus subtilis
0.236
-
 deoxycytidine pH 7.8, 25°C, wild-type enzyme Bacillus subtilis
0.267
-
 cytidine pH 7.6, 25°C, mutant enzyme R56Q Bacillus subtilis

Metals/Ions

Metals/Ions Comment Organism Structure
Zinc wild-type enzyme and mutant enzymes R56A, R56Q and C53H/R56Q contain about 1 mol of zinc per subunit. Mutant enzyme R56D contains 0.2 mol of zinc per subunit Bacillus subtilis

Organism

Organism UniProt Comment Textmining
Bacillus subtilis P19079
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Bacillus subtilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cytidine + H2O
-
 Bacillus subtilis uridine + NH3
-
 ?
deoxycytidine + H2O
-
 Bacillus subtilis deoxyuridine + NH3
-
 ?