BRENDA - Enzyme Database show

Recombinant human cytidine deaminase: Expression, purification, and characterization

Vincenzetti, S.; Cambi, A.; Neuhard, J.; Garattini, E.; Vita, A.; Protein Expr. Purif. 8, 247-253 (1996)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.5.4.5
cDNA, PCR, vector pTrc99A, cloned and expressed in Escherichia coli DH5alpha
Homo sapiens
Crystallization (Commentary)
EC Number
Crystallization
Organism
3.5.4.5
-
Escherichia coli
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.4.5
3,4,5,6-Tetrahydrouridine
recombinant CDA
Homo sapiens
3.5.4.5
3-deazauridine
recombinant enzyme
Homo sapiens
3.5.4.5
5-fluoro-2'-deoxyuridine
recombinant enzyme
Homo sapiens
3.5.4.5
5-{(1E)-3-[(5-carboxypentyl)amino]-3-oxoprop-1-en-1-yl}-5,6-dihydrocytidine
recombinant enzyme
Homo sapiens
3.5.4.5
CMP
recombinant CDA
Homo sapiens
3.5.4.5
thymine riboside
recombinant CDA
Homo sapiens
3.5.4.5
uridine
recombinant enzyme
Homo sapiens
3.5.4.5
uridine
-
Escherichia coli
3.5.4.5
UTP
recombinant CDA
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.4.5
0.039
-
cytidine
recombinant CDA
Homo sapiens
3.5.4.5
0.039
-
deoxycytidine
recombinant CDA
Homo sapiens
3.5.4.5
0.04
-
5-methylcytidine
recombinant CDA
Homo sapiens
3.5.4.5
0.1
-
5-iododeoxycytidine
recombinant CDA
Homo sapiens
3.5.4.5
0.11
-
cytosine arabinoside
recombinant CDA
Homo sapiens
3.5.4.5
0.28
-
5-azacytidine
recombinant CDA
Homo sapiens
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.4.5
15000
-
4 * 15000, SDS-PAGE, purified enzyme, homotetramer
Homo sapiens
3.5.4.5
16000
-
estimated from cDNA open reading frame coding for human cytidine deaminase
Homo sapiens
3.5.4.5
16170
-
deduced from cDNA nucleotide sequence
Homo sapiens
3.5.4.5
32000
-
2 * 32000, homodimer, SDS-PAGE
Escherichia coli
3.5.4.5
57000
-
gel filtration
Escherichia coli
3.5.4.5
57000
-
gel filtration
Homo sapiens
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.5.4.5
cytidine + H2O
Bacillus subtilis
-
uridine + NH3
-
-
?
3.5.4.5
cytidine + H2O
Escherichia coli
-
uridine + NH3
-
-
?
3.5.4.5
cytidine + H2O
Homo sapiens
-
uridine + NH3
-
-
?
3.5.4.5
cytidine + H2O
Escherichia coli DH5-alpha
-
uridine + NH3
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.5.4.5
Bacillus subtilis
-
-
-
3.5.4.5
Escherichia coli
-
strain DH5alpha
-
3.5.4.5
Escherichia coli DH5-alpha
-
strain DH5alpha
-
3.5.4.5
Homo sapiens
-
human
-
Purification (Commentary)
EC Number
Commentary
Organism
3.5.4.5
-
Homo sapiens
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
3.5.4.5
granulocyte
-
Homo sapiens
-
3.5.4.5
leukocyte
-
Homo sapiens
-
3.5.4.5
liver
-
Homo sapiens
-
3.5.4.5
lymphocyte
-
Homo sapiens
-
3.5.4.5
placenta
-
Homo sapiens
-
3.5.4.5
spleen
-
Homo sapiens
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.5.4.5
105
-
-
Homo sapiens
Storage Stability
EC Number
Storage Stability
Organism
3.5.4.5
-20°C, 20 mM Tris-HCl, pH 7.5, 1 mM DDT, 1 mM EDTA, 20% glycerol, enzyme retains full activity for at least 1 month, 80% of activity lost in absence of glycerol
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
1-beta-D-arabinofuranosylcytosine + H2O
-
209665
Escherichia coli
?
-
-
-
?
3.5.4.5
1-beta-D-arabinofuranosylcytosine + H2O
-
209665
Homo sapiens
?
-
-
-
?
3.5.4.5
1-beta-D-arabinofuranosylcytosine + H2O
-
209665
Escherichia coli DH5-alpha
?
-
-
-
?
3.5.4.5
5-azacytidine + H2O
-
209665
Homo sapiens
5-azauridine + NH3
-
-
-
-
3.5.4.5
5-azadeoxycytidine + H2O
-
209665
Homo sapiens
5-azadeoxyuridine + NH3
-
-
-
?
3.5.4.5
5-methylcytidine + H2O
-
209665
Homo sapiens
5-methyluridine + NH3
-
-
-
-
3.5.4.5
cytidine + H2O
-
209665
Bacillus subtilis
uridine + NH3
-
-
-
?
3.5.4.5
cytidine + H2O
-
209665
Escherichia coli
uridine + NH3
-
-
-
?
3.5.4.5
cytidine + H2O
-
209665
Homo sapiens
uridine + NH3
-
-
-
?
3.5.4.5
cytidine + H2O
-
209665
Escherichia coli DH5-alpha
uridine + NH3
-
-
-
?
3.5.4.5
deoxycytidine + H2O
-
209665
Escherichia coli
deoxyuridine + NH3
-
-
-
?
3.5.4.5
deoxycytidine + H2O
-
209665
Homo sapiens
deoxyuridine + NH3
-
-
-
?
3.5.4.5
deoxycytidine + H2O
-
209665
Escherichia coli DH5-alpha
deoxyuridine + NH3
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
3.5.4.5
dimer
2 * 32000, homodimer, SDS-PAGE
Escherichia coli
3.5.4.5
tetramer
4 * 15000, SDS-PAGE, purified enzyme, homotetramer; 4 * 16000, amino acid sequence
Homo sapiens
3.5.4.5
tetramer
-
Bacillus subtilis
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.5.4.5
3.5
10.5
deoxycytidine as substrate, beyond these values irreversible inactivation occurs
Homo sapiens
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.4.5
cDNA, PCR, vector pTrc99A, cloned and expressed in Escherichia coli DH5alpha
Homo sapiens
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
3.5.4.5
-
Escherichia coli
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.4.5
3,4,5,6-Tetrahydrouridine
recombinant CDA
Homo sapiens
3.5.4.5
3-deazauridine
recombinant enzyme
Homo sapiens
3.5.4.5
5-fluoro-2'-deoxyuridine
recombinant enzyme
Homo sapiens
3.5.4.5
5-{(1E)-3-[(5-carboxypentyl)amino]-3-oxoprop-1-en-1-yl}-5,6-dihydrocytidine
recombinant enzyme
Homo sapiens
3.5.4.5
CMP
recombinant CDA
Homo sapiens
3.5.4.5
thymine riboside
recombinant CDA
Homo sapiens
3.5.4.5
uridine
recombinant enzyme
Homo sapiens
3.5.4.5
uridine
-
Escherichia coli
3.5.4.5
UTP
recombinant CDA
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.4.5
0.039
-
cytidine
recombinant CDA
Homo sapiens
3.5.4.5
0.039
-
deoxycytidine
recombinant CDA
Homo sapiens
3.5.4.5
0.04
-
5-methylcytidine
recombinant CDA
Homo sapiens
3.5.4.5
0.1
-
5-iododeoxycytidine
recombinant CDA
Homo sapiens
3.5.4.5
0.11
-
cytosine arabinoside
recombinant CDA
Homo sapiens
3.5.4.5
0.28
-
5-azacytidine
recombinant CDA
Homo sapiens
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.4.5
15000
-
4 * 15000, SDS-PAGE, purified enzyme, homotetramer
Homo sapiens
3.5.4.5
16000
-
estimated from cDNA open reading frame coding for human cytidine deaminase
Homo sapiens
3.5.4.5
16170
-
deduced from cDNA nucleotide sequence
Homo sapiens
3.5.4.5
32000
-
2 * 32000, homodimer, SDS-PAGE
Escherichia coli
3.5.4.5
57000
-
gel filtration
Escherichia coli
3.5.4.5
57000
-
gel filtration
Homo sapiens
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.5.4.5
cytidine + H2O
Bacillus subtilis
-
uridine + NH3
-
-
?
3.5.4.5
cytidine + H2O
Escherichia coli
-
uridine + NH3
-
-
?
3.5.4.5
cytidine + H2O
Homo sapiens
-
uridine + NH3
-
-
?
3.5.4.5
cytidine + H2O
Escherichia coli DH5-alpha
-
uridine + NH3
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.4.5
-
Homo sapiens
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
3.5.4.5
granulocyte
-
Homo sapiens
-
3.5.4.5
leukocyte
-
Homo sapiens
-
3.5.4.5
liver
-
Homo sapiens
-
3.5.4.5
lymphocyte
-
Homo sapiens
-
3.5.4.5
placenta
-
Homo sapiens
-
3.5.4.5
spleen
-
Homo sapiens
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.5.4.5
105
-
-
Homo sapiens
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
3.5.4.5
-20°C, 20 mM Tris-HCl, pH 7.5, 1 mM DDT, 1 mM EDTA, 20% glycerol, enzyme retains full activity for at least 1 month, 80% of activity lost in absence of glycerol
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
1-beta-D-arabinofuranosylcytosine + H2O
-
209665
Escherichia coli
?
-
-
-
?
3.5.4.5
1-beta-D-arabinofuranosylcytosine + H2O
-
209665
Homo sapiens
?
-
-
-
?
3.5.4.5
1-beta-D-arabinofuranosylcytosine + H2O
-
209665
Escherichia coli DH5-alpha
?
-
-
-
?
3.5.4.5
5-azacytidine + H2O
-
209665
Homo sapiens
5-azauridine + NH3
-
-
-
-
3.5.4.5
5-azadeoxycytidine + H2O
-
209665
Homo sapiens
5-azadeoxyuridine + NH3
-
-
-
?
3.5.4.5
5-methylcytidine + H2O
-
209665
Homo sapiens
5-methyluridine + NH3
-
-
-
-
3.5.4.5
cytidine + H2O
-
209665
Bacillus subtilis
uridine + NH3
-
-
-
?
3.5.4.5
cytidine + H2O
-
209665
Escherichia coli
uridine + NH3
-
-
-
?
3.5.4.5
cytidine + H2O
-
209665
Homo sapiens
uridine + NH3
-
-
-
?
3.5.4.5
cytidine + H2O
-
209665
Escherichia coli DH5-alpha
uridine + NH3
-
-
-
?
3.5.4.5
deoxycytidine + H2O
-
209665
Escherichia coli
deoxyuridine + NH3
-
-
-
?
3.5.4.5
deoxycytidine + H2O
-
209665
Homo sapiens
deoxyuridine + NH3
-
-
-
?
3.5.4.5
deoxycytidine + H2O
-
209665
Escherichia coli DH5-alpha
deoxyuridine + NH3
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.5.4.5
dimer
2 * 32000, homodimer, SDS-PAGE
Escherichia coli
3.5.4.5
tetramer
4 * 15000, SDS-PAGE, purified enzyme, homotetramer; 4 * 16000, amino acid sequence
Homo sapiens
3.5.4.5
tetramer
-
Bacillus subtilis
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.5.4.5
3.5
10.5
deoxycytidine as substrate, beyond these values irreversible inactivation occurs
Homo sapiens