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Literature summary for 3.5.4.5 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)

Cloned (Comment) Organism
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

Protein Variants Comment Organism
F137A site-directed mutagenesis, inactive monomeric enzyme Homo sapiens
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
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
Y33F site-directed mutagenesis, inactive monomeric enzyme Homo sapiens
Y33G site-directed mutagenesis, inactive monomeric enzyme Homo sapiens
Y33S site-directed mutagenesis, inactive monomeric enzyme Homo sapiens
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

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

KM Value [mM]

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

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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)

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

Subunits

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

Synonyms

Synonyms Comment Organism
T-CDA
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
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
58
-
recombinant wild-type enzyme Homo sapiens

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
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
37 75 60% of maximal activity at 75°C for the wild-type enzyme, 35% for mutant R68Q Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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

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

Ki Value [mM]

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