Cloned (Comment) | Organism |
---|---|
expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
E239M | site-directed mutagenesis, the mutant shows reduced Vmax, increased Km, and lower molecular weight compared to the wild-type enzyme, subunit structure analysis | Bacillus subtilis |
E239Q | site-directed mutagenesis, the mutant shows reduced Vmax, increased Km, and lower molecular weight compared to the wild-type enzyme, subunit structure analysis | Bacillus subtilis |
E239R | site-directed mutagenesis, the mutant shows reduced Vmax, increased Km, and lower molecular weight compared to the wild-type enzyme, subunit structure analysis | Bacillus subtilis |
H299K | site-directed mutagenesis, the mutant shows reduced Vmax, increased Km, and lower molecular weight compared to the wild-type enzyme, subunit structure analysis | Bacillus subtilis |
H299R | site-directed mutagenesis, the mutant shows reduced Vmax, increased Km, and lower molecular weight compared to the wild-type enzyme, subunit structure analysis | Bacillus subtilis |
R167E | site-directed mutagenesis, the mutant shows reduced Vmax, increased Km, and lower molecular weight compared to the wild-type enzyme, subunit structure analysis | Bacillus subtilis |
R167Q | site-directed mutagenesis, the mutant shows reduced Vmax, increased Km, and lower molecular weight compared to the wild-type enzyme, subunit structure analysis | Bacillus subtilis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
KBr | presence of increasing concentrations of KBr of 0.12.5 M disrupt electrostatic interactions leading to ASL dissociation and loss of catalytic activity | Bacillus subtilis | |
additional information | as hydrophobic interactions are weakened at 8°C and 4°C, the catalytic activity of ASL decreases strikingly and the enzyme dissociates to a mixture of monomer-dimer-trimer, with small amounts of tetramer | Bacillus subtilis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-aminoimidazole-4-(N-succinylcarboxamide) ribonucleotide | Bacillus subtilis | - |
5-aminoimidazole-4-carboxamide ribonucleotide + fumarate | - |
? | |
succinyladenosine monophosphate | Bacillus subtilis | - |
AMP + fumarate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | P12047 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain Bl21 by nickel affinity chromatography | Bacillus subtilis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
specific activities of different enzyme variants with different subunit numbers | Bacillus subtilis |
0.2 | - |
wild-type enzyme at 4 mg/ml and 4°C | Bacillus subtilis |
0.4 | - |
wild-type enzyme at 4 mg/ml and 8°C | Bacillus subtilis |
1.7 | - |
wild-type enzyme at 4 mg/ml and 25°C | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-aminoimidazole-4-(N-succinylcarboxamide) ribonucleotide | - |
Bacillus subtilis | 5-aminoimidazole-4-carboxamide ribonucleotide + fumarate | - |
? | |
5-aminoimidazole-4-(N-succinylcarboxamide) ribonucleotide | i.e. SAICAR | Bacillus subtilis | 5-aminoimidazole-4-carboxamide ribonucleotide + fumarate | i.e. AICAR | ? | |
succinyladenosine monophosphate | - |
Bacillus subtilis | AMP + fumarate | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | a homotetramer in which three subunits contribute to each of four active sites, analytical ultracentrifugation, gel filtration, circular dichroism spectroscopy, and interaction analysis, overview. ASL is active and exists as 100% tetramer at 25°C, while at 8°C and 4°C, as hydrophobic interactions are weakened, the catalytic activity decreases strikingly and the enzyme dissociates to a mixture of monomer-dimer-trimer, with small amounts of tetramer. In the presence of increasing concentrations of KBr of 0.12.5 M, which disrupts electrostatic interactions, ASL is dissociated initially to monomerdimer, with small amounts of trimer-tetramer, and then the monomer species predominates along with small amounts of trimer-tetramer, analytical ultracentrifugation, homology modelling of wild-type and mutant enzymes, overview | Bacillus subtilis |
Synonyms | Comment | Organism |
---|---|---|
ASL | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Bacillus subtilis |