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Literature summary extracted from

  • Vincenzetti, S.; Quadrini, B.; Mariani, P.; De Sanctis, G.; Cammertoni, N.; Polzonetti, V.; Pucciarelli, S.; Natalini, P.; Vita, A.
    Modulation of human cytidine deaminase by specific amino acids involved in the intersubunit interactions (2007), Proteins, 70, 144-156.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.4.5 expression of wild-type and mutant enzymes in Escherichia coli, complementation of enzyme enzyme-deficient strain by the wild-type enzyme and mutants Y60G, R68Q, and R68G, not by F33 and F137A mutants Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
3.5.4.5 F137A site-directed mutagenesis, inactive monomeric enzyme Homo sapiens
3.5.4.5 R68G site-directed mutagenesis, the mutant R68G shows reduced activity compared to the wild-type enzyme and dissociates very easily in presence of small amounts of SDS Homo sapiens
3.5.4.5 R68Q site-directed mutagenesis, the mutant shows reduced activity and thermal stability compared to the wild-type enzyme, but the quaternary structure of R68Q is not affected by the mutation Homo sapiens
3.5.4.5 Y33F site-directed mutagenesis, inactive monomeric enzyme Homo sapiens
3.5.4.5 Y33G site-directed mutagenesis, inactive monomeric enzyme Homo sapiens
3.5.4.5 Y33S site-directed mutagenesis, inactive monomeric enzyme Homo sapiens
3.5.4.5 Y60G site-directed mutagenesis, the mutant shows a more compact quaternary structure with respect to the wild-type enzyme, and a slightly reduced activity as well as altered thermal stability Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.4.5 3,4,5,6-Tetrahydrouridine i.e. THU Homo sapiens
3.5.4.5 5-fluorozebularine
-
Homo sapiens
3.5.4.5 6-[3-(5-cytidyl)acryloylamino]hexanoic acid i.e. CV6 Homo sapiens
3.5.4.5 uridine product inhibition Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.4.5 additional information
-
additional information kinetics of wild-type and mutant enzymes Homo sapiens
3.5.4.5 0.023
-
2'-deoxycytidine pH 7.5, 37°C, recombinant mutant R68G Homo sapiens
3.5.4.5 0.031
-
cytidine pH 7.5, 37°C, recombinant mutant R68G Homo sapiens
3.5.4.5 0.039
-
cytidine pH 7.5, 37°C, recombinant wild-type enzyme Homo sapiens
3.5.4.5 0.039
-
2'-deoxycytidine pH 7.5, 37°C, recombinant wild-type enzyme Homo sapiens
3.5.4.5 0.051
-
2'-deoxycytidine pH 7.5, 37°C, recombinant mutant R68Q Homo sapiens
3.5.4.5 0.11
-
cytosine arabinoside pH 7.5, 37°C, recombinant wild-type enzyme Homo sapiens
3.5.4.5 0.118
-
cytidine pH 7.5, 37°C, recombinant mutant R68Q Homo sapiens
3.5.4.5 0.15
-
cytosine arabinoside pH 7.5, 37°C, recombinant mutant R68G Homo sapiens
3.5.4.5 0.19
-
cytosine arabinoside pH 7.5, 37°C, recombinant mutant R68Q Homo sapiens
3.5.4.5 0.27
-
cytidine pH 7.5, 37°C, recombinant mutant Y60G Homo sapiens
3.5.4.5 0.385
-
cytosine arabinoside pH 7.5, 37°C, recombinant mutant Y60G Homo sapiens
3.5.4.5 0.4
-
2'-deoxycytidine pH 7.5, 37°C, recombinant mutant Y60G Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
3.5.4.5 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.4.5 recombinant wild-type enzyme by affinity chromatography on a cytidine analog 6-[3,5(cytidyl) acryloylamino]hexanoic acid resin, recombinant inactive mutants by anion exchange chromatography, ultrafiltration, and gel filtration Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.4.5 2'-deoxycytidine + H2O
-
Homo sapiens 2'-deoxyuridine + NH3
-
?
3.5.4.5 cytidine + H2O
-
Homo sapiens uridine + NH3
-
?
3.5.4.5 cytosine arabinoside + H2O
-
Homo sapiens ?
-
?
3.5.4.5 additional information substrate specificity of wild-type and mutant enzymes, overview Homo sapiens ?
-
?

Subunits

EC Number Subunits Comment Organism
3.5.4.5 tetramer homotetramer, residue Y33 is involved in enzyme folding, enzyme structure analysis using circular dichroism, molecular modeling, overview Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
3.5.4.5 T-CDA
-
Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.4.5 30 62 mutant Y60G shows an unusual behavior with a mean of 75% activity for temperatures within this range and three peaks of maximum activity at about 30°C, 46°C, and 62°C Homo sapiens
3.5.4.5 58
-
recombinant wild-type enzyme Homo sapiens

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.5.4.5 15 65 mutant Y60G shows an unusual behavior with a mean of 75% activity for temperatures within this range and three peaks of maximum activity at about 30°C, 46°C, and 62°C Homo sapiens
3.5.4.5 37 75 60% of maximal activity at 75°C for the wild-type enzyme, 35% for mutant R68Q Homo sapiens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.4.5 15 75 wild-type enzyme and mutant R68Q are stable, while the stability of mutant Y60G is decreased above 65°C Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.4.5 7.5
-
assay at Homo sapiens

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.5.4.5 0.0003
-
5-fluorozebularine pH 7.5, 37°C, recombinant wild-type enzyme Homo sapiens
3.5.4.5 0.00042
-
3,4,5,6-Tetrahydrouridine pH 7.5, 37°C, recombinant mutant R68Q Homo sapiens
3.5.4.5 0.0006
-
5-fluorozebularine pH 7.5, 37°C, recombinant mutant R68Q Homo sapiens
3.5.4.5 0.0017
-
5-fluorozebularine pH 7.5, 37°C, recombinant mutant Y60G Homo sapiens
3.5.4.5 0.0067
-
3,4,5,6-Tetrahydrouridine pH 7.5, 37°C, recombinant wild-type enzyme Homo sapiens
3.5.4.5 0.008
-
3,4,5,6-Tetrahydrouridine pH 7.5, 37°C, recombinant mutant Y60G Homo sapiens
3.5.4.5 0.015
-
6-[3-(5-cytidyl)acryloylamino]hexanoic acid pH 7.5, 37°C, recombinant mutant Y60G Homo sapiens
3.5.4.5 0.045
-
6-[3-(5-cytidyl)acryloylamino]hexanoic acid pH 7.5, 37°C, recombinant wild-type enzyme Homo sapiens
3.5.4.5 0.1
-
uridine pH 7.5, 37°C, recombinant mutant Y60G Homo sapiens
3.5.4.5 0.12
-
uridine pH 7.5, 37°C, recombinant mutant R68Q Homo sapiens
3.5.4.5 0.4
-
uridine pH 7.5, 37°C, recombinant wild-type enzyme Homo sapiens