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Literature summary 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

Cloned(Commentary)

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

General Stability

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.4.2 0.032
-
 adenine
-
 Schizosaccharomyces pombe
3.5.4.2 0.032
-
 adenine 0.2 M phosphate buffer, 33°C Schizosaccharomyces pombe
3.5.4.2 0.055
-
 adenine
-
 Saccharomyces cerevisiae
3.5.4.2 0.055
-
 adenine 0.2 M phosphate buffer, 33°C Saccharomyces cerevisiae
3.5.4.4 0.032
-
 adenine
-
 Schizosaccharomyces pombe

Localization

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.4.2 39630
-
 calculated Saccharomyces cerevisiae
3.5.4.2 39630
-
 deduced from sequence Saccharomyces cerevisiae
3.5.4.2 41200
-
 calculated Schizosaccharomyces pombe
3.5.4.2 41200
-
 deduced from sequence Schizosaccharomyces pombe
3.5.4.2 46000
-
 SDS-PAGE Saccharomyces cerevisiae
3.5.4.2 48000
-
 SDS-PAGE Schizosaccharomyces pombe
3.5.4.4 39630
-
 calculated Saccharomyces cerevisiae
3.5.4.4 39950
-
 calculated Arabidopsis thaliana
3.5.4.4 41200
-
 calculated Schizosaccharomyces pombe

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.4.2 adenine + H2O Schizosaccharomyces pombe
-
 hypoxanthine + NH3
-
 ir
3.5.4.2 adenine + H2O Saccharomyces cerevisiae
-
 hypoxanthine + NH3
-
 ir
3.5.4.2 adenine + H2O Saccharomyces cerevisiae irreversible deamination, metabolic salvage pathway of purines hypoxanthine + NH3
-
 ir
3.5.4.2 adenine + H2O Schizosaccharomyces pombe irreversible deamination, metabolic salvage pathway of purines hypoxanthine + NH3
-
 ir
3.5.4.2 kinetin + H2O Saccharomyces cerevisiae release of hypoxanthine and inosine, release of amine from from the cytokinine side chain ?
-
 ?
3.5.4.2 kinetin + H2O Schizosaccharomyces pombe release of hypoxanthine and inosine, release of amine from from the cytokinine side chain ?
-
 ?
3.5.4.2 N6-(2-isopentenyl)adenine + H2O Schizosaccharomyces pombe partial conversion, at endo-side of N6 amino groups, release of N6-side chain amines ?
-
 ?
3.5.4.2 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

EC Number Organism UniProt Comment Textmining
3.5.4.2 Saccharomyces cerevisiae
-
-
-
3.5.4.2 Saccharomyces cerevisiae P53909 this enzyme catalyses the reaction of EC 3.5.4.2. and EC 3.5.4.4; strain 23344c
-
3.5.4.2 Schizosaccharomyces pombe
-
-
-
3.5.4.2 Schizosaccharomyces pombe Q9P6I7 this enzyme catalyses the reaction of EC 3.5.4.2 and EC 3.5.4.4; wild-type 972 h
-
3.5.4.4 Arabidopsis thaliana Q8LPL7
-
-
3.5.4.4 Saccharomyces cerevisiae P53909 this enzyme catalyses the reaction of EC 3.5.4.2 and EC 3.5.4.4; strain 23344c
-
3.5.4.4 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)

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.4.2 additional information
-
 139 nkat/mg with substrate adenine Saccharomyces cerevisiae
3.5.4.2 additional information
-
 326 nkat/mg with substrate adenine Schizosaccharomyces pombe
3.5.4.2 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
3.5.4.2 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
3.5.4.4 additional information
-
 326 nkat/mg with substrate adenine, reaction of EC 3.5.4.2, adenine deaminase Schizosaccharomyces pombe
3.5.4.4 0.12
-
 0.033 mM adenosine in 0.2 M phosphate buffer (pH 6.7), 30°C Arabidopsis thaliana

Storage Stability

EC Number Storage Stability Organism
3.5.4.2 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
3.5.4.2 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
3.5.4.2 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
3.5.4.2 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
3.5.4.4 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
3.5.4.4 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
3.5.4.4 activity of purified proteins 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 Arabidopsis thaliana

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.4.2 6-[(E)-4-hydroxy-3-methylbut-2-enylamino]purine + H2O 1.8% of the activity of adenine Saccharomyces cerevisiae ?
-
 ?
3.5.4.2 6-[(Z)-4-hydroxy-3-methylbut-2-enylamino]purine + H2O 1.7% of the activity of adenine Saccharomyces cerevisiae ?
-
 ?
3.5.4.2 adenine + H2O
-
 Schizosaccharomyces pombe hypoxanthine + NH3
-
 ir
3.5.4.2 adenine + H2O
-
 Saccharomyces cerevisiae hypoxanthine + NH3
-
 ir
3.5.4.2 adenine + H2O irreversible deamination, metabolic salvage pathway of purines Saccharomyces cerevisiae hypoxanthine + NH3
-
 ir
3.5.4.2 adenine + H2O irreversible deamination, metabolic salvage pathway of purines Schizosaccharomyces pombe hypoxanthine + NH3
-
 ir
3.5.4.2 benzyladenine + H2O 0.7% of the activity of adenine Saccharomyces cerevisiae hypoxanthine + 1-phenylmethaneamine
-
 ir
3.5.4.2 benzyladenine + H2O 3.8% of the activity of adenine Schizosaccharomyces pombe hypoxanthine + 1-phenylmethaneamine
-
 ir
3.5.4.2 kinetin + H2O release of hypoxanthine and inosine, release of amine from from the cytokinine side chain Saccharomyces cerevisiae ?
-
 ?
3.5.4.2 kinetin + H2O release of hypoxanthine and inosine, release of amine from from the cytokinine side chain Schizosaccharomyces pombe ?
-
 ?
3.5.4.2 kinetin + H2O hydrolysis of adenine compounds, cleavage of cytokinins into inosine and the corresponding side chain amine Saccharomyces cerevisiae ?
-
 ?
3.5.4.2 kinetin + H2O hydrolysis of adenine compounds, cleavage of cytokinins into inosine and the corresponding side chain amine Schizosaccharomyces pombe ?
-
 ?
3.5.4.2 kinetin + H2O 2.6% of the activity of adenine Saccharomyces cerevisiae hypoxanthine + 1-furan-2-ylmethaneamine
-
 ir
3.5.4.2 kinetin + H2O 2.8% of the activity of adenine Schizosaccharomyces pombe hypoxanthine + 1-furan-2-ylmethaneamine
-
 ir
3.5.4.2 m-topolin + H2O 2.5% of the activity of adenine Saccharomyces cerevisiae hypoxanthine + 2-(aminomethyl)phenol
-
 ir
3.5.4.2 m-topolin + H2O 4.0% of the activity of adenine Schizosaccharomyces pombe hypoxanthine + 2-(aminomethyl)phenol
-
 ir
3.5.4.2 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 ?
-
 ?
3.5.4.2 N6-(2-isopentenyl)adenine + H2O partial conversion, at endo-side of N6 amino groups, release of N6-side chain amines Schizosaccharomyces pombe ?
-
 ?
3.5.4.2 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 ?
-
 ?
3.5.4.2 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
-
 ?
3.5.4.2 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
-
 ?
3.5.4.2 N6-isopentenyladenine + H2O 2.1% of the activity of adenine Saccharomyces cerevisiae hypoxanthine + 3-methylbut-3-en-1-amine
-
 ir
3.5.4.2 N6-isopentenyladenine + H2O 2.9% of the activity of adenine Schizosaccharomyces pombe hypoxanthine + 3-methylbut-3-en-1-amine
-
 ir
3.5.4.4 adenine + H2O
-
 Schizosaccharomyces pombe hypoxanthine + NH3
-
 ?
3.5.4.4 adenine + H2O
-
 Arabidopsis thaliana ? + NH3
-
 ?
3.5.4.4 adenosine + H2O 1.4% of the activity of adenine Saccharomyces cerevisiae inosine + NH3
-
 ?
3.5.4.4 adenosine + H2O 2.0% of the activity of adenine Schizosaccharomyces pombe inosine + NH3
-
 ?
3.5.4.4 adenosine + H2O 30% of the activity of adenine Arabidopsis thaliana inosine + NH3
-
 ir
3.5.4.4 adenosine 3'-monophosphate + H2O 56% of the activity of adenine Arabidopsis thaliana inosine 3'-monophosphate + NH3
-
 ?
3.5.4.4 adenosine 3'-phosphate + H2O 3.3% of the activity of adenine Saccharomyces cerevisiae inosine 3'-phosphate + NH3
-
 ?
3.5.4.4 adenosine 3'-phosphate + H2O 4.5% of the activity of adenine Schizosaccharomyces pombe inosine 3'-phosphate + NH3
-
 ?
3.5.4.4 ATP + H2O 3.3% of the activity of adenine Schizosaccharomyces pombe ITP + NH3
-
 ?
3.5.4.4 ATP + H2O 3.3% of the activity of adenine Saccharomyces cerevisiae ITP + NH3
-
 ?
3.5.4.4 ATP + H2O 68% of the activity of adenine Arabidopsis thaliana ITP + NH3
-
 ?
3.5.4.4 additional information does not cleave cytokinins Arabidopsis thaliana ?
-
 ?
3.5.4.4 N6-isopentenyladenosine + H2O 1.9% of the activity of adenine Schizosaccharomyces pombe ?
-
 ?
3.5.4.4 N6-isopentenyladenosine + H2O 1.2% of the activity of adenine Saccharomyces cerevisiae inosine + 3-methylbut-3-en-1-amine
-
 ?

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
3.5.4.2 adenine deaminase
-
 Schizosaccharomyces pombe
3.5.4.2 adenine deaminase
-
 Saccharomyces cerevisiae
3.5.4.4 ADA
-
 Arabidopsis thaliana
3.5.4.4 adenosine deaminase
-
 Arabidopsis thaliana

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.4.2 30 37 0.2 M phosphate buffer, pH 7.0 Saccharomyces cerevisiae
3.5.4.2 33
-
 assay at Schizosaccharomyces pombe
3.5.4.2 33
-
 0.2 M phosphate buffer, pH 6.7 Schizosaccharomyces pombe
3.5.4.2 35
-
 ranges from 30-37°C Saccharomyces cerevisiae
3.5.4.4 30
-
 0.2 M phosphate buffer, pH 6.7 Arabidopsis thaliana
3.5.4.4 30 37 0.2 M phosphate buffer, pH 7.0 Saccharomyces cerevisiae
3.5.4.4 33
-
 0.2 M phosphate buffer, pH 6.7 Schizosaccharomyces pombe

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.5.4.2 30 37 determined at pH 7.0 Saccharomyces cerevisiae

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.4.2 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
3.5.4.2 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.4.2 5.5
-
 adenine
-
 Saccharomyces cerevisiae
3.5.4.2 5.5
-
 adenine 0.067 mM adenine in 0.2 M phosphate buffer, 33°C, pH 7.0 Saccharomyces cerevisiae
3.5.4.2 13.4
-
 adenine
-
 Schizosaccharomyces pombe
3.5.4.4 0.005
-
 adenosine
-
 Arabidopsis thaliana

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.4.2 6.7
-
 assay at Schizosaccharomyces pombe
3.5.4.2 6.7
-
 0.2 M phosphate buffer, 33°C Schizosaccharomyces pombe
3.5.4.2 7
-
 assay at Saccharomyces cerevisiae
3.5.4.2 7
-
 0.2 M phosphate buffer, 30-37°C Saccharomyces cerevisiae
3.5.4.4 6.7
-
 0.2 M phosphate buffer, 30°C Arabidopsis thaliana
3.5.4.4 6.7
-
 0.2 M phosphate buffer, 33°C Schizosaccharomyces pombe
3.5.4.4 7
-
 0.2 M phosphate buffer, 30-37°C Saccharomyces cerevisiae

pH Stability

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