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Literature summary for 3.5.4.2 extracted from

  • Pospisilova, H.; Sebela, M.; Novak, O.; Frebort, I.
    Hydrolytic cleavage of N6-substituted adenine derivatives by eukaryotic adenine and adenosine deaminases (2008), Biosci. Rep., 28, 335-347.
    View publication on PubMed
Search for more literature for 3.5.4.2

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli, pDrive, pet100/D-TOPO, pet151/D-TOPO vectors Saccharomyces cerevisiae
expressed in Escherichia coli, pDrive, pet100/D-TOPO, pet151/D-TOPO vectors Schizosaccharomyces pombe
overexpression in Escherichia coli BL21 (DE3) Schizosaccharomyces pombe
overexpression in Escherichia coli BL21 (DE3) Saccharomyces cerevisiae

General Stability

General Stability Organism
after dissolving the freeze-dried enzyme a significant decrease od activity is observed within 3-4 h Saccharomyces cerevisiae
after dissolving the freeze-dried enzyme a significant decrease od activity is observed within 3-4 h Schizosaccharomyces pombe

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.032
-
 adenine
-
 Schizosaccharomyces pombe
0.032
-
 adenine 0.2 M phosphate buffer, 33°C Schizosaccharomyces pombe
0.055
-
 adenine
-
 Saccharomyces cerevisiae
0.055
-
 adenine 0.2 M phosphate buffer, 33°C Saccharomyces cerevisiae

Localization

Localization Comment Organism GeneOntology No. Textmining
intracellular
-
 Saccharomyces cerevisiae 5622
-
intracellular
-
 Schizosaccharomyces pombe 5622
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
39630
-
 calculated Saccharomyces cerevisiae
39630
-
 deduced from sequence Saccharomyces cerevisiae
41200
-
 calculated Schizosaccharomyces pombe
41200
-
 deduced from sequence Schizosaccharomyces pombe
46000
-
 SDS-PAGE Saccharomyces cerevisiae
48000
-
 SDS-PAGE Schizosaccharomyces pombe

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
adenine + H2O Schizosaccharomyces pombe
-
 hypoxanthine + NH3
-
 ir
adenine + H2O Saccharomyces cerevisiae
-
 hypoxanthine + NH3
-
 ir
adenine + H2O Saccharomyces cerevisiae irreversible deamination, metabolic salvage pathway of purines hypoxanthine + NH3
-
 ir
adenine + H2O Schizosaccharomyces pombe irreversible deamination, metabolic salvage pathway of purines hypoxanthine + NH3
-
 ir
kinetin + H2O Saccharomyces cerevisiae release of hypoxanthine and inosine, release of amine from from the cytokinine side chain ?
-
 ?
kinetin + H2O Schizosaccharomyces pombe release of hypoxanthine and inosine, release of amine from from the cytokinine side chain ?
-
 ?
N6-(2-isopentenyl)adenine + H2O Schizosaccharomyces pombe partial conversion, at endo-side of N6 amino groups, release of N6-side chain amines ?
-
 ?
N6-(2-isopentenyl)adenine + H2O Saccharomyces cerevisiae partial conversion, at endo-side of N6 amino groups, release of N6-side chain amines hypoxanthine + 3-methylbut-2-en-1-amine
-
 ?

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae
-
-
-
Saccharomyces cerevisiae P53909 this enzyme catalyses the reaction of EC 3.5.4.2. and EC 3.5.4.4; strain 23344c
-
Schizosaccharomyces pombe
-
-
-
Schizosaccharomyces pombe Q9P6I7 this enzyme catalyses the reaction of EC 3.5.4.2 and EC 3.5.4.4; wild-type 972 h
-

Purification (Commentary)

Purification (Comment) Organism
gel filtration, SDS-PAGE Saccharomyces cerevisiae
gel filtration, SDS-PAGE Schizosaccharomyces pombe
using Ni2+-nitrilotriacetate (Ni2+-NTA) column chromatography Schizosaccharomyces pombe
using Ni2+-nitrilotriacetate (Ni2+-NTA) column chromatography Saccharomyces cerevisiae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 139 nkat/mg with substrate adenine Saccharomyces cerevisiae
additional information
-
 326 nkat/mg with substrate adenine Schizosaccharomyces pombe
additional information
-
 comparable molecular properties with mammalian enzymes, adenine most favoured substrate, also hydrolyzes N6-substituted adenines from the cytokinin group of plant hormones, reaction products identified by mass spectroscopy, relative reaction rates given Schizosaccharomyces pombe
8.34
-
 recombinant protein, adenine as substrate, 0.067 mM in 0.2 M phosphate buffer, 33°C, pH 7.0, adenine most favoured substrate, also hydrolyzes N6-substituted adenines from the cytokinin group of plant hormones, reaction products identified by mass spectroscopy, relative reaction rates given Saccharomyces cerevisiae

Storage Stability

Storage Stability Organism
activity of purified protein is stable for 1-2 weeks when stored at 4°C at concentrations above 10 mg/ml with the addition of 20% (v/v) glycerol. Prolonged storage in solution is possible at -20°C with 50% (v/v) glycerol and at -80°C with 10-20% (v/v) glycerol or as freeze-dried powder Schizosaccharomyces pombe
activity of purified protein is stable for 1-2 weeks when stored at 4°C at concentrations above 10 mg/ml with the addition of 20% (v/v) glycerol. Prolonged storage in solution is possible at -20°C with 50% (v/v) glycerol and at -80°C with 10-20% (v/v) glycerol or as freeze-dried powder Saccharomyces cerevisiae
at 4°C in potassium buffer above a concentration of 10 mg/ml and in presence of 20% glycerol stable for 1-2 weeks, prolonged storage in solution at -20°C in presence of 50% glycerol and at -80°C in presence of 10-20% glycerol, or as freeze-dried powder Saccharomyces cerevisiae
at 4°C in potassium buffer above a concentration of 10 mg/ml and in presence of 20% glycerol stable for 1-2 weeks, prolonged storage in solution at -20°C in presence of 50% glycerol and at -80°C in presence of 10-20% glycerol, or as freeze-dried powder Schizosaccharomyces pombe

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6-[(E)-4-hydroxy-3-methylbut-2-enylamino]purine + H2O 1.8% of the activity of adenine Saccharomyces cerevisiae ?
-
 ?
6-[(Z)-4-hydroxy-3-methylbut-2-enylamino]purine + H2O 1.7% of the activity of adenine Saccharomyces cerevisiae ?
-
 ?
adenine + H2O
-
 Schizosaccharomyces pombe hypoxanthine + NH3
-
 ir
adenine + H2O
-
 Saccharomyces cerevisiae hypoxanthine + NH3
-
 ir
adenine + H2O irreversible deamination, metabolic salvage pathway of purines Saccharomyces cerevisiae hypoxanthine + NH3
-
 ir
adenine + H2O irreversible deamination, metabolic salvage pathway of purines Schizosaccharomyces pombe hypoxanthine + NH3
-
 ir
benzyladenine + H2O 0.7% of the activity of adenine Saccharomyces cerevisiae hypoxanthine + 1-phenylmethaneamine
-
 ir
benzyladenine + H2O 3.8% of the activity of adenine Schizosaccharomyces pombe hypoxanthine + 1-phenylmethaneamine
-
 ir
kinetin + H2O release of hypoxanthine and inosine, release of amine from from the cytokinine side chain Saccharomyces cerevisiae ?
-
 ?
kinetin + H2O release of hypoxanthine and inosine, release of amine from from the cytokinine side chain Schizosaccharomyces pombe ?
-
 ?
kinetin + H2O hydrolysis of adenine compounds, cleavage of cytokinins into inosine and the corresponding side chain amine Saccharomyces cerevisiae ?
-
 ?
kinetin + H2O hydrolysis of adenine compounds, cleavage of cytokinins into inosine and the corresponding side chain amine Schizosaccharomyces pombe ?
-
 ?
kinetin + H2O 2.6% of the activity of adenine Saccharomyces cerevisiae hypoxanthine + 1-furan-2-ylmethaneamine
-
 ir
kinetin + H2O 2.8% of the activity of adenine Schizosaccharomyces pombe hypoxanthine + 1-furan-2-ylmethaneamine
-
 ir
m-topolin + H2O 2.5% of the activity of adenine Saccharomyces cerevisiae hypoxanthine + 2-(aminomethyl)phenol
-
 ir
m-topolin + H2O 4.0% of the activity of adenine Schizosaccharomyces pombe hypoxanthine + 2-(aminomethyl)phenol
-
 ir
additional information the protein exhibits partial amino acid sequence similarity to the CHASE domain of the cytokinin receptor, which may account for its ability to bind and hydrolyse not only kinetin, but also other cytokinins and other cytokinin derivatives, albeit at lower rates than its natural substrate adenine Schizosaccharomyces pombe ?
-
 ?
N6-(2-isopentenyl)adenine + H2O partial conversion, at endo-side of N6 amino groups, release of N6-side chain amines Schizosaccharomyces pombe ?
-
 ?
N6-(2-isopentenyl)adenine + H2O hydrolysis of adenine compounds, low activity towards N6-substituted adenines, partial conversion, release of N6-side chain amines Schizosaccharomyces pombe ?
-
 ?
N6-(2-isopentenyl)adenine + H2O partial conversion, at endo-side of N6 amino groups, release of N6-side chain amines Saccharomyces cerevisiae hypoxanthine + 3-methylbut-2-en-1-amine
-
 ?
N6-(2-isopentenyl)adenine + H2O hydrolysis of adenine compounds, low activity towards N6-substituted adenines, partial conversion, release of N6-side chain amines Saccharomyces cerevisiae hypoxanthine + 3-methylbut-2-en-1-amine
-
 ?
N6-isopentenyladenine + H2O 2.1% of the activity of adenine Saccharomyces cerevisiae hypoxanthine + 3-methylbut-3-en-1-amine
-
 ir
N6-isopentenyladenine + H2O 2.9% of the activity of adenine Schizosaccharomyces pombe hypoxanthine + 3-methylbut-3-en-1-amine
-
 ir

Subunits

Subunits Comment Organism
? x * 48000, SDS-PAGE Schizosaccharomyces pombe
? x * 46000, SDS-PAGE Saccharomyces cerevisiae

Synonyms

Synonyms Comment Organism
adenine deaminase
-
 Schizosaccharomyces pombe
adenine deaminase
-
 Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30 37 0.2 M phosphate buffer, pH 7.0 Saccharomyces cerevisiae
33
-
 assay at Schizosaccharomyces pombe
33
-
 0.2 M phosphate buffer, pH 6.7 Schizosaccharomyces pombe
35
-
 ranges from 30-37°C Saccharomyces cerevisiae

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 37 determined at pH 7.0 Saccharomyces cerevisiae

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 prolonged storage in solution, at -20°C in presence of 50% glycerol, at -80°C in presence of 10-20% glycerol Saccharomyces cerevisiae
additional information
-
 prolonged storage in solution, at -20°C in presence of 50% glycerol, at -80°C in presence of 10-20% glycerol Schizosaccharomyces pombe

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.5
-
 adenine
-
 Saccharomyces cerevisiae
5.5
-
 adenine 0.067 mM adenine in 0.2 M phosphate buffer, 33°C, pH 7.0 Saccharomyces cerevisiae
13.4
-
 adenine
-
 Schizosaccharomyces pombe

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.7
-
 assay at Schizosaccharomyces pombe
6.7
-
 0.2 M phosphate buffer, 33°C Schizosaccharomyces pombe
7
-
 assay at Saccharomyces cerevisiae
7
-
 0.2 M phosphate buffer, 30-37°C Saccharomyces cerevisiae

pH Stability

pH Stability pH Stability Maximum Comment Organism
additional information
-
 activity highest in potassium phosphate buffer, decreasing activity to 75% in Tris/HCl buffer and to 30% in MOPS buffer Saccharomyces cerevisiae
additional information
-
 activity highest in potassium phosphate buffer, decreasing activity to 75% in Tris/HCl buffer and to 30% in MOPS buffer Schizosaccharomyces pombe