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 | 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 | 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
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 | 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 | Comment | Organism |
---|---|---|
tetramer | homotetramer, residue Y33 is involved in enzyme folding, enzyme structure analysis using circular dichroism, molecular modeling, overview | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
T-CDA | - |
Homo sapiens |
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 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 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Homo sapiens |
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 |