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Literature summary extracted from

  • Minici, C.; Cacciapuoti, G.; De Leo, E.; Porcelli, M.; Degano, M.
    New determinants in the catalytic mechanism of nucleoside hydrolases from the structures of two isozymes from Sulfolobus solfataricus (2012), Biochemistry, 51, 4590-4599.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.2.1 overexpressed in Escherichia coli Saccharolobus solfataricus
3.2.2.8 overexpressed in Escherichia coli Saccharolobus solfataricus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.2.1 crystal structure is determined at 1.8 A resolution. It is crystallized using the hanging drop vapor diffusion method, mixing 0.001 ml of protein and 0.001 ml of aprecipitant solution consisting of 100 mM Hepes (pH 7.5), 5% PEG 3350, 5 mM CaCl2, 5 mM CdCl2, and 5 mM MgCl2 Saccharolobus solfataricus
3.2.2.8 crystal structure is determined at 1.6 A resolution. The enzyme is crystallized using the hanging drop vapor diffusion method by mixing an equal amount of protein and a precipitant solution, composed of 100 mM bicine (pH 9) and 1.5 M ammonium sulfate Saccharolobus solfataricus

Protein Variants

EC Number Protein Variants Comment Organism
3.2.2.1 H79A kcat/Km for inosine is 13fold lower than wild-type value Saccharolobus solfataricus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.2.1 0.33
-
 Inosine mutant enzyme H79A, pH and temperature not specified in the publication Saccharolobus solfataricus
3.2.2.1 0.34
-
 Inosine wild-type enzyme, pH and temperature not specified in the publication Saccharolobus solfataricus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.2.1 Ca2+ the active site contains the trademark Ca2+ ion of nucleodide hydrolase enzymes, octacoordinated by residues Asp9, Asp14, Ile121, and Asp238, two solvent molecules, and one glycerol molecule Saccharolobus solfataricus
3.2.2.8 Na+ likely bound to the enzyme under physiological conditions Saccharolobus solfataricus

Organism

EC Number Organism UniProt Comment Textmining
3.2.2.1 Saccharolobus solfataricus Q97WH6
-
-
3.2.2.1 Saccharolobus solfataricus P2 Q97WH6
-
-
3.2.2.8 Saccharolobus solfataricus Q97ZS5
-
-
3.2.2.8 Saccharolobus solfataricus P2 Q97ZS5
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.2.1
-
 Saccharolobus solfataricus
3.2.2.8
-
 Saccharolobus solfataricus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.2.1 inosine + H2O
-
 Saccharolobus solfataricus hypoxanthine + D-ribose
-
 ?
3.2.2.1 inosine + H2O
-
 Saccharolobus solfataricus P2 hypoxanthine + D-ribose
-
 ?
3.2.2.8 inosine + H2O
-
 Saccharolobus solfataricus hypoxanthine + D-ribose
-
 ?
3.2.2.8 inosine + H2O
-
 Saccharolobus solfataricus P2 hypoxanthine + D-ribose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.2.1 purine-preferring nucleoside hydrolase
-
 Saccharolobus solfataricus
3.2.2.1 purine-specific inosine-adenosine-guanosine nucleoside hydrolase
-
 Saccharolobus solfataricus
3.2.2.1 SsIAG-NH
-
 Saccharolobus solfataricus
3.2.2.1 SSO2243 locus name Saccharolobus solfataricus
3.2.2.8 pyrimidine-specific nucleoside hydrolase
-
 Saccharolobus solfataricus
3.2.2.8 SsCU-NH
-
 Saccharolobus solfataricus
3.2.2.8 SSO0505
-
 Saccharolobus solfataricus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.2.1 0.85
-
 Inosine mutant enzyme H79A, pH and temperature not specified in the publication Saccharolobus solfataricus
3.2.2.1 11.6
-
 Inosine wild-type enzyme, pH and temperature not specified in the publication Saccharolobus solfataricus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.2.1 2.6
-
 Inosine mutant enzyme H79A, pH and temperature not specified in the publication Saccharolobus solfataricus
3.2.2.1 34.1
-
 Inosine wild-type enzyme, pH and temperature not specified in the publication Saccharolobus solfataricus