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Literature summary 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

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.5.4.5 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

EC Number Protein Variants Comment Organism
3.5.4.5 C53H attempts to express and purify the mutant enzyme are unsuccessful Bacillus subtilis
3.5.4.5 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
3.5.4.5 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
3.5.4.5 R56D zinc-binding capacity of mutant enzyme is reduced Bacillus subtilis
3.5.4.5 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.4.5 0.0075
-
 deoxycytidine pH 7.8, 25°C, mutant enzyme R56A Bacillus subtilis
3.5.4.5 0.029
-
 deoxycytidine pH 7.8, 25°C, mutant enzyme R56Q Bacillus subtilis
3.5.4.5 0.14
-
 cytidine pH 7.6, 25°C, mutant enzyme R56A Bacillus subtilis
3.5.4.5 0.216
-
 cytidine pH 7.6, 25°C, wild-type enzyme Bacillus subtilis
3.5.4.5 0.236
-
 deoxycytidine pH 7.8, 25°C, wild-type enzyme Bacillus subtilis
3.5.4.5 0.267
-
 cytidine pH 7.6, 25°C, mutant enzyme R56Q Bacillus subtilis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.4.5 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

EC Number Organism UniProt Comment Textmining
3.5.4.5 Bacillus subtilis P19079
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.4.5
-
 Bacillus subtilis

Substrates and Products (Substrate)

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