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Literature summary for 2.4.2.1 extracted from
- Biochemical and structural characterization of mammalian-like purine nucleoside phosphorylase from the Archaeon Pyrococcus furiosus (2007), FEBS J., 274, 2482-2495.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli | Pyrococcus furiosus |
| expression in Escherichia coli | Pyrococcus furiosus |
Protein Variants
| 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
| 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 [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 [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 Stability
| 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
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pyrococcus furiosus | - |
- |
- |
| Pyrococcus furiosus | Q8U2I1 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Pyrococcus furiosus |
| recombinant | Pyrococcus furiosus |
Specific Activity [micromol/min/mg]
| 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 and Products (Substrate)
| 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
| 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
| Synonyms | Comment | Organism |
|---|---|---|
| PfPNP | - |
Pyrococcus furiosus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 120 | - |
- |
Pyrococcus furiosus |
Temperature Range [°C]
| 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 [°C]
| 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 [1/s]
| 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 | |
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