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Literature summary for 3.2.2.1 extracted from

  • Porcelli, M.; Peluso, I.; Marabotti, A.; Facchiano, A.; Cacciapuoti, G.
    Biochemical characterization and homology modeling of a purine-specific ribonucleoside hydrolase from the archaeon Sulfolobus solfataricus: insights into mechanisms of protein stabilization (2009), Arch. Biochem. Biophys., 483, 55-65.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Saccharolobus solfataricus

General Stability

General Stability Organism
IAG-NH is characterized by remarkable resistance to guanidinium chloride-induced unfolding Saccharolobus solfataricus

Inhibitors

Inhibitors Comment Organism Structure
EDTA after 1 h incubation with EDTA (5 mM) the enzyme activity is reduced to less than 0.05% and is restored by addition of 20 mM CaCl2 Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.06
-
adenosine at 80°C Saccharolobus solfataricus
0.16
-
guanosine at 80°C Saccharolobus solfataricus
0.36
-
Inosine at 80°C Saccharolobus solfataricus

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ IAG-NH is a Ca2+-dependent enzyme Saccharolobus solfataricus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
34325
-
2 * 34325, calculated from sequence of cDNA Saccharolobus solfataricus
35000
-
subunit, SDS-PAGE Saccharolobus solfataricus
68650
-
calculated from sequence of cDNA Saccharolobus solfataricus

Organic Solvent Stability

Organic Solvent Comment Organism
dithiothreitol after 1 h pre-incubation at temperatures until 80°C, the enzyme remains completely stable even at high concentrations of dithiothreitol (0.7 M) whereas it becomes susceptible to the effect of the reducing agent as the temperature raises, after 1 h pre-incubation in the presence of 0.5 M, 0.6 M and 0.7 M dithiothreitol, IAG-NH retains 70%, 60% and 21% activity respectively Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus
-
-
-
Saccharolobus solfataricus P2
-
-
-

Purification (Commentary)

Purification (Comment) Organism
methylthioinosine-Sepharose column chromatography Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
adenosine + H2O IAG-NH shows higher affinity for adenosine with respect to inosine and guanosine, but the catalytic efficiency of these substrates is comparable Saccharolobus solfataricus D-ribose + adenine
-
?
guanosine + H2O highest activity Saccharolobus solfataricus D-ribose + guanine
-
?
guanosine + H2O highest activity Saccharolobus solfataricus P2 D-ribose + guanine
-
?
inosine + H2O
-
Saccharolobus solfataricus D-ribose + hypoxanthine
-
?
additional information IAG-NH does not recognize as substrate nucleosides 5'-phosphates, NAD+ and NADP+ Saccharolobus solfataricus ?
-
?
additional information IAG-NH does not recognize as substrate nucleosides 5'-phosphates, NAD+ and NADP+ Saccharolobus solfataricus P2 ?
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 34325, calculated from sequence of cDNA Saccharolobus solfataricus

Synonyms

Synonyms Comment Organism
IAG-NH the enzyme is specific for adenosine, guanosine and inosine Saccharolobus solfataricus
inosine-adenosine-guanosine nucleoside hydrolase
-
Saccharolobus solfataricus
purine-specific ribonucleoside hydrolase
-
Saccharolobus solfataricus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
105
-
-
Saccharolobus solfataricus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
60 115 the enzyme is highly thermoactive, its activity increases sharply up to the optimal temperature of 105°C and 50% activity is still observed at 115°C Saccharolobus solfataricus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
100 110 IAG-NH is characterized by extreme thermodynamic stability with a melting temperature at 107°C, it retains full activity after 1 h incubation at 90°C and shows half-lives of 45 min, 30 min and 15 min at 100°C, 105°C, and 110°C, respectively Saccharolobus solfataricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.8
-
adenosine at 80°C Saccharolobus solfataricus
23.3
-
Inosine at 80°C Saccharolobus solfataricus
29.1
-
guanosine at 80°C Saccharolobus solfataricus

pI Value

Organism Comment pI Value Maximum pI Value
Saccharolobus solfataricus estimated isoelectric point
-
5.9