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Literature summary for 6.3.4.18 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
Search for more literature for 6.3.4.18

Cloned(Commentary)

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

Protein Variants

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

Inhibitors

Inhibitors Comment Organism Structure
ATP substrate inhibition Aspergillus clavatus

KM Value [mM]

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

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Aspergillus clavatus A1CII2
-
-
Aspergillus clavatus DSM 816 A1CII2
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

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

Turnover Number [1/s]

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

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Aspergillus clavatus

Ki Value [mM]

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

kcat/KM [mM/s]

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