BRENDA - Enzyme Database show

Modulation of human cytidine deaminase by specific amino acids involved in the intersubunit interactions

Vincenzetti, S.; Quadrini, B.; Mariani, P.; De Sanctis, G.; Cammertoni, N.; Polzonetti, V.; Pucciarelli, S.; Natalini, P.; Vita, A.; Proteins 70, 144-156 (2007)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
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
Engineering
EC Number
Amino acid exchange
Commentary
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
Commentary
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
Commentary
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
-
2'-deoxycytidine
pH 7.5, 37°C, recombinant wild-type enzyme
Homo sapiens
3.5.4.5
0.039
-
cytidine
pH 7.5, 37°C, recombinant wild-type enzyme
Homo sapiens
3.5.4.5
0.051
-
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
Primary Accession No. (UniProt)
Commentary
Textmining
3.5.4.5
Homo sapiens
-
-
-
Purification (Commentary)
EC Number
Commentary
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
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.4.5
2'-deoxycytidine + H2O
-
689935
Homo sapiens
2'-deoxyuridine + NH3
-
-
-
?
3.5.4.5
cytidine + H2O
-
689935
Homo sapiens
uridine + NH3
-
-
-
?
3.5.4.5
cytosine arabinoside + H2O
-
689935
Homo sapiens
?
-
-
-
?
3.5.4.5
additional information
substrate specificity of wild-type and mutant enzymes, overview
689935
Homo sapiens
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
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
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
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]
Commentary
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]
Commentary
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
Commentary
Organism
3.5.4.5
7.5
-
assay at
Homo sapiens
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
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
Cloned(Commentary) (protein specific)
EC Number
Commentary
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
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
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 (protein specific)
EC Number
Inhibitors
Commentary
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
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
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
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
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
-
2'-deoxycytidine
pH 7.5, 37°C, recombinant wild-type enzyme
Homo sapiens
3.5.4.5
0.039
-
cytidine
pH 7.5, 37°C, recombinant wild-type enzyme
Homo sapiens
3.5.4.5
0.051
-
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
Purification (Commentary) (protein specific)
EC Number
Commentary
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) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.4.5
2'-deoxycytidine + H2O
-
689935
Homo sapiens
2'-deoxyuridine + NH3
-
-
-
?
3.5.4.5
cytidine + H2O
-
689935
Homo sapiens
uridine + NH3
-
-
-
?
3.5.4.5
cytosine arabinoside + H2O
-
689935
Homo sapiens
?
-
-
-
?
3.5.4.5
additional information
substrate specificity of wild-type and mutant enzymes, overview
689935
Homo sapiens
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
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
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
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] (protein specific)
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
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] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
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 (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.5.4.5
7.5
-
assay at
Homo sapiens