Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Pyrococcus furiosus |
expression in Escherichia coli | Pyrococcus furiosus |
Protein Variants | Comment | Organism |
---|---|---|
C254S/C256S | reduced thermodynamic and kinetic stability of the mutant with respect to the wild-type PfPNP | Pyrococcus furiosus |
C254S/C256S | Tm-value is 8°C lower than Tm-value of wild-type enzyme | Pyrococcus furiosus |
General Stability | Organism |
---|---|
limited proteolysis indicated that the only proteolytic cleavage site is localized in the C-terminal region | Pyrococcus furiosus |
phosphate is able to increase the already high stability of PfPNP toward SDS | Pyrococcus furiosus |
remarkable SDS-resistance, enzyme remains fully active after 30 min in 2% SDS at 50°C and retains 60% residual activity after 5 min at 90°C | Pyrococcus furiosus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.122 | - |
guanosine | 80°C | Pyrococcus furiosus | |
0.322 | - |
Inosine | 80°C | Pyrococcus furiosus | |
0.322 | - |
Inosine | 80°C, pH 7.4 | Pyrococcus furiosus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
28977 | - |
6 * 28977, calculated from sequence | Pyrococcus furiosus |
29000 | - |
6 * 29000, SDS-PAGE | Pyrococcus furiosus |
29208 | - |
6 * 29208, calculated from sequence | Pyrococcus furiosus |
180000 | - |
gel filtration | Pyrococcus furiosus |
Organic Solvent | Comment | Organism |
---|---|---|
SDS | 2%, the enzyme remains fully active after 30 min incubation at 50°C and still retains 60% residual activity after 5 min incubation at 90°C. Phosphate is able to increase the already high stability of PfPNP toward the detergent | Pyrococcus furiosus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus furiosus | - |
- |
- |
Pyrococcus furiosus | Q8U2I1 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Pyrococcus furiosus |
recombinant | Pyrococcus furiosus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.0179 | - |
- |
Pyrococcus furiosus |
0.0179 | - |
80°C, production of hypoxanthine | Pyrococcus furiosus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
guanosine + phosphate | - |
Pyrococcus furiosus | guanine + alpha-D-ribose 1-phosphate | - |
? | |
guanosine + phosphate | absolute specificity for inosine and guanosine | Pyrococcus furiosus | guanine + alpha-D-ribose 1-phosphate | - |
? | |
inosine + phosphate | - |
Pyrococcus furiosus | hypoxanthine + alpha-D-ribose 1-phosphate | - |
? | |
inosine + phosphate | absolute specificity for inosine and guanosine | Pyrococcus furiosus | hypoxanthine + alpha-D-ribose 1-phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | 6 * 28977, calculated from sequence | Pyrococcus furiosus |
hexamer | 6 * 29000, SDS-PAGE | Pyrococcus furiosus |
hexamer | 6 * 29208, calculated from sequence | Pyrococcus furiosus |
Synonyms | Comment | Organism |
---|---|---|
PfPNP | - |
Pyrococcus furiosus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
120 | - |
- |
Pyrococcus furiosus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
75 | 133 | 75°C: about 55% of maximal activity, 133°C: about 50% of maximal activity | Pyrococcus furiosus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
three pairs of intrasubunit disulfide bridges play a role in the stability of the enzyme against thermal inactivation | Pyrococcus furiosus |
100 | - |
4 h, stable | Pyrococcus furiosus |
100 | - |
4 h, stable, wild-type enzyme. Mutant enzyme C254S/C256S shows 38% residual activity after 4 h incubation | Pyrococcus furiosus |
102 | - |
Tm-value for mutant enzyme C254S/C256S | Pyrococcus furiosus |
105 | - |
half-life: 69 min | Pyrococcus furiosus |
105 | - |
half-life: 69 min for wild-type enzyme, 35.5 min for mutant enzyme C254S/C256S | Pyrococcus furiosus |
110 | - |
Tm-value is 110°C, half-life: 12 min | Pyrococcus furiosus |
110 | - |
half-life: 12 min, Tm-value 110°C increases to 120°C in the presence of 100 mM phosphate, wild-type enzyme | Pyrococcus furiosus |
115 | - |
half-life: 5 min | Pyrococcus furiosus |
115 | - |
half-life: 5 min, wild-type enzyme | Pyrococcus furiosus |
120 | - |
Tm-value in presence of 100 mM phosphate | Pyrococcus furiosus |
120 | - |
Tm-value 110°C increases to 120°C in the presence of 100 mM phosphate, wild-type enzyme | Pyrococcus furiosus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
28.05 | - |
guanosine | 80°C | Pyrococcus furiosus | |
84.19 | - |
Inosine | 80°C | Pyrococcus furiosus |