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Literature summary extracted from

  • Dewal, M.; Firestine, S.
    Site-directed mutagenesis of catalytic residues in N5-carboxyaminoimidazole ribonucleotide synthetase (2013), Biochemistry, 52, 6559-6567.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.4.18 expressed in Escherichia coli BL21(DE3) pLysS cells Aspergillus clavatus

Protein Variants

EC Number Protein Variants Comment Organism
6.3.4.18 D153A the mutant shows wild type activity Aspergillus clavatus
6.3.4.18 E73A the mutant shows 2fold reduced activity compared to the wild type enzyme Aspergillus clavatus
6.3.4.18 H273A inactive Aspergillus clavatus
6.3.4.18 H273Q inactive Aspergillus clavatus
6.3.4.18 K353A inactive Aspergillus clavatus
6.3.4.18 K353R inactive Aspergillus clavatus
6.3.4.18 R155A the mutant shows wild type activity Aspergillus clavatus
6.3.4.18 R271A inactive Aspergillus clavatus
6.3.4.18 R271K the mutant shows almost no (1000fold) reducedactivity compared to the wild type enzyme and is more sensitive towards ATP substrate inhibition Aspergillus clavatus
6.3.4.18 R271Q inactive Aspergillus clavatus
6.3.4.18 Y152A the mutant shows 3fold reduced activity compared to the wild type enzyme Aspergillus clavatus
6.3.4.18 Y152F the mutant shows wild type activity Aspergillus clavatus

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.4.18 ATP substrate inhibition Aspergillus clavatus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.4.18 2.8
-
 HCO3- mutant enzyme Y152F, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 3.9
-
 HCO3- wild type enzyme, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 5
-
 HCO3- mutant enzyme Y152A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 7
-
 HCO3- mutant enzyme R155A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 15
-
 HCO3- mutant enzyme D153A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 33
-
 HCO3- mutant enzyme E73A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 80
-
 HCO3- mutant enzyme R271K, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.4.18 ATP + 5-amino-1-(5-phospho-D-ribosyl)imidazole + HCO3- + H+ Aspergillus clavatus
-
 ADP + phosphate + 5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole
-
 ?
6.3.4.18 ATP + 5-amino-1-(5-phospho-D-ribosyl)imidazole + HCO3- + H+ Aspergillus clavatus DSM 816
-
 ADP + phosphate + 5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.4.18 Aspergillus clavatus A1CII2
-
-
6.3.4.18 Aspergillus clavatus DSM 816 A1CII2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.4.18 HisPur cobalt resin column chromatography Aspergillus clavatus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.4.18 ATP + 5-amino-1-(5-phospho-D-ribosyl)imidazole + HCO3- + H+
-
 Aspergillus clavatus ADP + phosphate + 5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole
-
 ?
6.3.4.18 ATP + 5-amino-1-(5-phospho-D-ribosyl)imidazole + HCO3- + H+
-
 Aspergillus clavatus DSM 816 ADP + phosphate + 5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.3.4.18 N5-CAIR synthetase
-
 Aspergillus clavatus
6.3.4.18 N5-carboxyaminoimidazole ribonucleotide synthetase
-
 Aspergillus clavatus
6.3.4.18 PurK
-
 Aspergillus clavatus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.3.4.18 0.0013
-
 HCO3- mutant enzyme R271K, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 0.032
-
 HCO3- mutant enzyme Y152A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 0.053
-
 HCO3- mutant enzyme Y152F, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 0.063
-
 HCO3- wild type enzyme, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 0.143
-
 HCO3- mutant enzyme R155A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 0.233
-
 HCO3- mutant enzyme D153A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 0.25
-
 HCO3- mutant enzyme E73A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.4.18 ATP
-
 Aspergillus clavatus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
6.3.4.18 1.2
-
 ATP mutant enzyme R271K, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 3.3
-
 ATP wild type enzyme, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.3.4.18 0.017
-
 HCO3- mutant enzyme R271K, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 6.7
-
 HCO3- mutant enzyme Y152A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 8.3
-
 HCO3- mutant enzyme E73A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 15
-
 HCO3- mutant enzyme D153A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 16.7
-
 HCO3- mutant enzyme R155A, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 16.7
-
 HCO3- mutant enzyme Y152F, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus
6.3.4.18 16.7
-
 HCO3- wild type enzyme, in 50 mM HEPES, pH 7.8, at 37°C Aspergillus clavatus