Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Cacciapuoti, G.; Gorassini, S.; Mazzeo, M.F.; Siciliano, R.A.; Carbone, V.; Zappia, V.; Porcelli, M.
    Biochemical and structural characterization of mammalian-like purine nucleoside phosphorylase from the Archaeon Pyrococcus furiosus (2007), FEBS J., 274, 2482-2495.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.1 expressed in Escherichia coli Pyrococcus furiosus
2.4.2.1 expression in Escherichia coli Pyrococcus furiosus

Protein Variants

EC Number Protein Variants Comment Organism
2.4.2.1 C254S/C256S reduced thermodynamic and kinetic stability of the mutant with respect to the wild-type PfPNP Pyrococcus furiosus
2.4.2.1 C254S/C256S Tm-value is 8°C lower than Tm-value of wild-type enzyme Pyrococcus furiosus

General Stability

EC Number General Stability Organism
2.4.2.1 limited proteolysis indicated that the only proteolytic cleavage site is localized in the C-terminal region Pyrococcus furiosus
2.4.2.1 phosphate is able to increase the already high stability of PfPNP toward SDS Pyrococcus furiosus
2.4.2.1 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.2.1 0.122
-
guanosine 80°C Pyrococcus furiosus
2.4.2.1 0.322
-
Inosine 80°C Pyrococcus furiosus
2.4.2.1 0.322
-
Inosine 80°C, pH 7.4 Pyrococcus furiosus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.2.1 28977
-
6 * 28977, calculated from sequence Pyrococcus furiosus
2.4.2.1 29000
-
6 * 29000, SDS-PAGE Pyrococcus furiosus
2.4.2.1 29208
-
6 * 29208, calculated from sequence Pyrococcus furiosus
2.4.2.1 180000
-
gel filtration Pyrococcus furiosus

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
2.4.2.1 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

EC Number Organism UniProt Comment Textmining
2.4.2.1 Pyrococcus furiosus
-
-
-
2.4.2.1 Pyrococcus furiosus Q8U2I1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.2.1
-
Pyrococcus furiosus
2.4.2.1 recombinant Pyrococcus furiosus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.2.1 0.0179
-
-
Pyrococcus furiosus
2.4.2.1 0.0179
-
80°C, production of hypoxanthine Pyrococcus furiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.1 guanosine + phosphate
-
Pyrococcus furiosus guanine + alpha-D-ribose 1-phosphate
-
?
2.4.2.1 guanosine + phosphate absolute specificity for inosine and guanosine Pyrococcus furiosus guanine + alpha-D-ribose 1-phosphate
-
?
2.4.2.1 inosine + phosphate
-
Pyrococcus furiosus hypoxanthine + alpha-D-ribose 1-phosphate
-
?
2.4.2.1 inosine + phosphate absolute specificity for inosine and guanosine Pyrococcus furiosus hypoxanthine + alpha-D-ribose 1-phosphate
-
?

Subunits

EC Number Subunits Comment Organism
2.4.2.1 hexamer 6 * 28977, calculated from sequence Pyrococcus furiosus
2.4.2.1 hexamer 6 * 29000, SDS-PAGE Pyrococcus furiosus
2.4.2.1 hexamer 6 * 29208, calculated from sequence Pyrococcus furiosus

Synonyms

EC Number Synonyms Comment Organism
2.4.2.1 PfPNP
-
Pyrococcus furiosus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.2.1 120
-
-
Pyrococcus furiosus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.4.2.1 75 133 75°C: about 55% of maximal activity, 133°C: about 50% of maximal activity Pyrococcus furiosus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.2.1 additional information
-
three pairs of intrasubunit disulfide bridges play a role in the stability of the enzyme against thermal inactivation Pyrococcus furiosus
2.4.2.1 100
-
4 h, stable Pyrococcus furiosus
2.4.2.1 100
-
4 h, stable, wild-type enzyme. Mutant enzyme C254S/C256S shows 38% residual activity after 4 h incubation Pyrococcus furiosus
2.4.2.1 102
-
Tm-value for mutant enzyme C254S/C256S Pyrococcus furiosus
2.4.2.1 105
-
half-life: 69 min Pyrococcus furiosus
2.4.2.1 105
-
half-life: 69 min for wild-type enzyme, 35.5 min for mutant enzyme C254S/C256S Pyrococcus furiosus
2.4.2.1 110
-
Tm-value is 110°C, half-life: 12 min Pyrococcus furiosus
2.4.2.1 110
-
half-life: 12 min, Tm-value 110°C increases to 120°C in the presence of 100 mM phosphate, wild-type enzyme Pyrococcus furiosus
2.4.2.1 115
-
half-life: 5 min Pyrococcus furiosus
2.4.2.1 115
-
half-life: 5 min, wild-type enzyme Pyrococcus furiosus
2.4.2.1 120
-
Tm-value in presence of 100 mM phosphate Pyrococcus furiosus
2.4.2.1 120
-
Tm-value 110°C increases to 120°C in the presence of 100 mM phosphate, wild-type enzyme Pyrococcus furiosus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.2.1 28.05
-
guanosine 80°C Pyrococcus furiosus
2.4.2.1 84.19
-
Inosine 80°C Pyrococcus furiosus