Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3)/pSJS1244 cells | Mycoplasma pneumoniae |
Protein Variants | Comment | Organism |
---|---|---|
D124A | inactive | Mycoplasma pneumoniae |
D151A | the mutant shows increased turnover number compared to the wild type enzyme | Mycoplasma pneumoniae |
D154A | inactive | Mycoplasma pneumoniae |
D81A | the mutant shows reduced turnover number compared to the wild type enzyme | Mycoplasma pneumoniae |
K120A | the mutant shows increased turnover number compared to the wild type enzyme | Mycoplasma pneumoniae |
K3A | the mutant shows strongly reduced turnover number compared to the wild type enzyme | Mycoplasma pneumoniae |
K3D/D151K | the mutant shows strongly reduced turnover number compared to the wild type enzyme | Mycoplasma pneumoniae |
K3D/D154 K | inactive | Mycoplasma pneumoniae |
R125A | the mutant shows reduced turnover number compared to the wild type enzyme | Mycoplasma pneumoniae |
R7A | the mutant shows severely reduced turnover number compared to the wild type enzyme | Mycoplasma pneumoniae |
W153A | the mutant shows reduced turnover number compared to the wild type enzyme | Mycoplasma pneumoniae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.076 | - |
ATP | wild type enzyme, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
0.086 | - |
ATP | mutant enzyme D81A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
0.15 | - |
ATP | mutant enzyme D151A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
1 | 4 | 5-formyltetrahydrofolate | mutant enzyme D151A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
1.1 | - |
ATP | mutant enzyme K3D/D151K, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
1.2 | - |
ATP | mutant enzyme K120A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
2.5 | - |
5-formyltetrahydrofolate | wild type enzyme, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
5 | - |
ATP | mutant enzyme K3A, Km above 5 mM, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
5 | - |
ATP | mutant enzyme R125A, Km above 5 mM, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
5 | - |
ATP | mutant enzyme R7A, Km above 5 mM, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
5 | - |
ATP | mutant enzyme W153A, Km above 5 mM, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
5.8 | - |
5-formyltetrahydrofolate | mutant enzyme D81A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
7.1 | - |
5-formyltetrahydrofolate | mutant enzyme K120A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
21 | - |
5-formyltetrahydrofolate | mutant enzyme W153A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
31 | - |
5-formyltetrahydrofolate | mutant enzyme R7A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
51 | - |
5-formyltetrahydrofolate | mutant enzyme K3A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
75 | - |
5-formyltetrahydrofolate | mutant enzyme R125A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
130 | - |
5-formyltetrahydrofolate | mutant enzyme K3D/D151K, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 5-formyltetrahydrofolate | Mycoplasma pneumoniae | - |
ADP + phosphate + 5,10-methenyltetrahydrofolate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycoplasma pneumoniae | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Talon metal affinity column chromatography | Mycoplasma pneumoniae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 5-formyltetrahydrofolate | - |
Mycoplasma pneumoniae | ADP + phosphate + 5,10-methenyltetrahydrofolate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
5,10-Methenyltetrahydrofolate synthetase | - |
Mycoplasma pneumoniae |
MTHFS | - |
Mycoplasma pneumoniae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.046 | - |
5-formyltetrahydrofolate | mutant enzyme R7A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
0.1 | - |
5-formyltetrahydrofolate | mutant enzyme K3D/D151K, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
0.17 | - |
5-formyltetrahydrofolate | mutant enzyme K3A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
0.35 | - |
5-formyltetrahydrofolate | mutant enzyme R125A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
0.77 | - |
5-formyltetrahydrofolate | mutant enzyme D81A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
0.8 | - |
5-formyltetrahydrofolate | mutant enzyme W153A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
1 | - |
5-formyltetrahydrofolate | wild type enzyme, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
1.3 | - |
5-formyltetrahydrofolate | mutant enzyme D151A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae | |
1.7 | - |
5-formyltetrahydrofolate | mutant enzyme K120A, in 200 mM MES (pH 6.0), 30 mM sodium chloride, 10 mM magnesium chloride, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol at 37°C | Mycoplasma pneumoniae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Mycoplasma pneumoniae |