Activating Compound | Comment | Organism | Structure |
---|---|---|---|
5-hydroxymethyl-dCTP | natural positive allosteric effector, enzyme is much more effectively regulated by its natural effector, 5-hydroxymethyl-dCTP, than by dCTP, binding of 5-hydroxymethyl-dCTP is much more pH dependent than dCTP | Tequatrovirus T4 | |
5-hydroxymethyl-dCTP | natural positive allosteric effector, enzyme is much more effectively regulated by its natural effector, 5-hydroxymethyl-dCTP, than by dCTP, binding of 5-hydroxymethyl-dCTP is much more pH dependent than dCTP | Escherichia phage T2 | |
5-hydroxymethyl-dCTP | natural positive allosteric effector, enzyme is much more effectively regulated by its natural effector, 5-hydroxymethyl-dCTP, than by dCTP, binding of 5-hydroxymethyl-dCTP is much more pH dependent than dCTP | Enterobacteria phage T6 | |
5-hydroxymethyl-dCTP | natural activator, required for activity | Tequatrovirus T4 | |
dCTP | required for activity | Tequatrovirus T4 | |
dCTP | positive allosteric effector | Tequatrovirus T4 | |
dCTP | positive allosteric effector | Escherichia phage T2 | |
dCTP | positive allosteric effector | Enterobacteria phage T6 | |
dCTP | hexameric form of enzyme is activated by dCTP, while the dimer is not | Tequatrovirus T4 | |
dCTP | wild-type, dCTP required for activity, no activity in absence of dCTP and Mg2+ | Tequatrovirus T4 | |
dCTP | severalfold activation, activation requires presence of Mg2+ | Tequatrovirus T4 | |
dCTP | severalfold activation, activation requires presence of Mg2+ | Escherichia phage T2 | |
dCTP | severalfold activation, activation requires presence of Mg2+ | Enterobacteria phage T6 | |
dCTP | mutants R115E and R115Q: dCTP not required. R115Q: slight, 30-40% activation by 0.02 mM dCTP | Tequatrovirus T4 | |
additional information | the phage dCMP deaminase expression is increased upon infection of Escherichia coli | Tequatrovirus T4 |
Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Tequatrovirus T4 |
Protein Variants | Comment | Organism |
---|---|---|
F112A | molecular weight analysis using HPLC gel filtration in the presence of SDS and dCTP: wild-type is a hexamer, F112A varies from hexamer to dimer | Tequatrovirus T4 |
F112A | site-directed mutagenesis, the mutant enzyme shows reduced turnover and activity compared to the wild-type enzyme | Tequatrovirus T4 |
R115E | mutant enzyme active in the absence of dCTP and Mg2+, little if any activation by dCTP, possessing turnover number or kcat that is about 15% that of wild-type enzyme, specific activity about 40-50% of wild-type enzyme. Molecular weight analysis using HPLC gel filtration in the presence of SDS and dCTP: wild-type is a hexamer, R115E a dimer | Tequatrovirus T4 |
R115E | site-directed mutagenesis, the mutant enzyme shows reduced turnover and activity compared to the wild-type enzyme, but no longer requires deoxycytidine 5-triphosphate for activation in contrast to the wild-type enzyme | Tequatrovirus T4 |
R115Q | mutant enzyme active in the absence of dCTP and Mg2+, possessing turnover number or kcat that is about 15% that of wild-type enzyme, specific activity about 40-50% of wild-type enzyme. Molecular weight analysis using HPLC size gel filtration in the presence of SDS and dCTP: wild-type is a hexamer, R115Q varies from hexamer to dimer | Tequatrovirus T4 |
R115Q | site-directed mutagenesis, the mutant enzyme shows reduced turnover and activity compared to the wild-type enzyme, but no longer requires deoxycytidine 5-triphosphate for activation in contrast to the wild-type enzyme | Tequatrovirus T4 |
General Stability | Organism |
---|---|
2-mercaptoethanol stabilizes | Tequatrovirus T4 |
2-mercaptoethanol stabilizes | Escherichia phage T2 |
EDTA: denaturation, EDTA removes the two resident Zn2+ atoms /subunit | Tequatrovirus T4 |
guanidine-HCl, 6 M, denaturation, complete restoration of activity on removal of denaturant by dilution or dialysis | Tequatrovirus T4 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
dTTP | allosteric inhibitor | Enterobacteria phage T6 | |
dTTP | allosteric inhibitor | Escherichia phage T2 | |
dTTP | allosteric inhibitor; wild-type, 0.1 mM dTTP: 20% inhibition, mutants R115E and R115Q: 0.1 mM dTTP: no inhibition. 0.3 mM dTTP: wild-type 90% inhibition, mutants R115E and R115Q: 30% inhibition | Tequatrovirus T4 | |
EDTA | - |
Tequatrovirus T4 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | - |
Escherichia phage T2 | |
additional information | - |
additional information | kinetic analysis, data and characterization of wild-type and mutants R115E, R115Q and F112A | Tequatrovirus T4 | |
0.021 | - |
dCMP | wild-type, presence of dCTP and Mg2+ | Tequatrovirus T4 | |
0.021 | - |
dCMP | recombinant wild-type enzyme, pH 8.0, 30°C | Tequatrovirus T4 | |
0.127 | - |
dCMP | R115Q, absence of dCTP | Tequatrovirus T4 | |
0.127 | - |
dCMP | recombinant mutant R115Q, pH 8.0, 30°C | Tequatrovirus T4 | |
0.137 | - |
dCMP | R115E, absence of dCTP | Tequatrovirus T4 | |
0.137 | - |
dCMP | recombinant mutant R115E, pH 8.0, 30°C | Tequatrovirus T4 |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Tequatrovirus T4 | |
Mg2+ | required for activation by dCTP and for inhibition by dTTP | Tequatrovirus T4 | |
Mg2+ | required for activation by dCTP and for inhibition by dTTP | Escherichia phage T2 | |
Mg2+ | required for activation by dCTP and for inhibition by dTTP | Enterobacteria phage T6 | |
Zn2+ | two resident Zn2+ atoms /subunit | Tequatrovirus T4 |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
21200 | - |
6 * 21200, wild-type, particularly in the presence of dCTP: hexamer, mutants R115Q and F112A varies from hexamers to dimers or something in between depending on protein concentration, HPLC gel filtration in the presence of SDS | Tequatrovirus T4 |
21200 | - |
2 * 21200, mutant R115E: dimer, mutants R115Q and F112A varies from hexamers to dimers or something in between depending on protein concentration, HPLC gel filtration in the presence of SDS | Tequatrovirus T4 |
38200 | 113000 | recombinant mutant F112A, gel filtration, dependent on protein concentration, in presence and absence of fdCTP | Tequatrovirus T4 |
39800 | 118000 | recombinant mutant R115Q, gel filtration, dependent on protein concentration, in presence and absence of fdCTP | Tequatrovirus T4 |
41600 | 45400 | recombinant mutant R115E, gel filtration, dependent on protein concentration, in presence and absence of fdCTP | Tequatrovirus T4 |
44300 | - |
R115E, HPLC gel filtration | Tequatrovirus T4 |
110000 | 122000 | recombinant wild-type enzyme, gel filtration, dependent on protein concentration, in presence and absence of fdCTP | Tequatrovirus T4 |
111900 | - |
F112A, HPLC gel filtration | Tequatrovirus T4 |
124500 | - |
R115Q, HPLC gel filtration | Tequatrovirus T4 |
127000 | - |
wild-type, HPLC gel filtration | Tequatrovirus T4 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
dCMP + H2O | Tequatrovirus T4 | - |
dUMP + NH3 | - |
? | |
dCMP + H2O | Escherichia phage T2 | - |
dUMP + NH3 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Enterobacteria phage T6 | - |
T2-, T4-, and T6-phage induced enzyme in Escherichia coli | - |
Escherichia phage T2 | - |
T2-, T4-, and T6-phage induced enzyme in Escherichia coli | - |
Tequatrovirus T4 | - |
- |
- |
Tequatrovirus T4 | - |
T2-, T4-, and T6-phage induced enzyme in Escherichia coli | - |
Tequatrovirus T4 | - |
wild-type and R115E and R115Q, mutant enzymes that no longer requires dCTP for activation, and mutant F112A | - |
Tequatrovirus T4 | - |
T4-phage infected Escherichia coli | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli | Tequatrovirus T4 |
wild-type and mutants R115E, R115Q and F112A | Tequatrovirus T4 |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
dCMP + H2O = dUMP + NH3 | regulation, allosteric end-product regulation | Tequatrovirus T4 | |
dCMP + H2O = dUMP + NH3 | regulation, allosteric end-product regulation | Escherichia phage T2 | |
dCMP + H2O = dUMP + NH3 | regulation, allosteric end-product regulation | Enterobacteria phage T6 | |
dCMP + H2O = dUMP + NH3 | highly regulated allosteric enzyme | Tequatrovirus T4 | |
dCMP + H2O = dUMP + NH3 | highly regulated allosteric enzyme | Escherichia phage T2 | |
dCMP + H2O = dUMP + NH3 | highly regulated allosteric enzyme | Enterobacteria phage T6 |
Renatured (Comment) | Organism |
---|---|
after denaturation with EDTA, mutants R115E and R115Q restored 54% and 60% of original activities, wild-type enzyme only marginally restored | Tequatrovirus T4 |
after treatment with guanidine-HCl, 6 M, complete reactivation on removal of denaturant by dilution or dialysis | Tequatrovirus T4 |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
mutants R115E and R115Q possessing specific activity about 40-50% of wild-type enzyme | Tequatrovirus T4 |
90 | 100 | mutant R115E | Tequatrovirus T4 |
121 | - |
mutant R115Q | Tequatrovirus T4 |
220 | - |
wild-type | Tequatrovirus T4 |
Storage Stability | Organism |
---|---|
-40°C, solid ammonium sulfate to 80% saturation, stable for longer periods of storage | Tequatrovirus T4 |
0-4°C, 0.1 M 2-mercaptoethanol, stable for extended periods | Tequatrovirus T4 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dCMP + H2O | - |
Tequatrovirus T4 | dUMP + NH3 | - |
? | |
dCMP + H2O | - |
Escherichia phage T2 | dUMP + NH3 | - |
? | |
dCMP + H2O | - |
Enterobacteria phage T6 | dUMP + NH3 | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 21200, mutant R115E: dimer, mutants R115Q and F112A varies from hexamers to dimers or something in between depending on protein concentration, HPLC gel filtration in the presence of SDS | Tequatrovirus T4 |
dimer | mutants R115Q and R115E, mutant F112A changes between dimeric and hexameric forms | Tequatrovirus T4 |
hexamer | 6 identical subunits | Tequatrovirus T4 |
hexamer | 6 identical subunits | Escherichia phage T2 |
hexamer | 6 identical subunits | Enterobacteria phage T6 |
hexamer | 6 * 21200, wild-type, particularly in the presence of dCTP: hexamer, mutants R115Q and F112A varies from hexamers to dimers or something in between depending on protein concentration, HPLC gel filtration in the presence of SDS | Tequatrovirus T4 |
hexamer | wild-type enzyme, mutant F112A changes between dimeric and hexameric forms | Tequatrovirus T4 |
More | hexameric form of enzyme is activated by dCTP, while the dimer is not | Tequatrovirus T4 |
Synonyms | Comment | Organism |
---|---|---|
dCMP deaminase | - |
Tequatrovirus T4 |
deoxycytidylate deaminase | - |
Tequatrovirus T4 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Tequatrovirus T4 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | mutants R115E and R115Q possessing turnover number or kcat that is about 15% that of wild-type enzyme | Tequatrovirus T4 | |
69 | - |
dCMP | R115E | Tequatrovirus T4 | |
69 | - |
dCMP | mutant R115E, pH 8.0, 30°C | Tequatrovirus T4 | |
85 | - |
dCMP | R115Q dimer | Tequatrovirus T4 | |
85 | - |
dCMP | mutant R115Q, pH 8.0, 30°C | Tequatrovirus T4 | |
255 | - |
dCMP | R115Q hexamer | Tequatrovirus T4 | |
466 | - |
dCMP | wild-type | Tequatrovirus T4 | |
466 | - |
dCMP | wild-type enzyme, pH 8.0, 30°C | Tequatrovirus T4 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Tequatrovirus T4 |
8 | - |
assay at | Escherichia phage T2 |