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Literature summary extracted from

  • Tolley, M.; Bickford, L.; Clare, K.; Johann, T.W.
    Investigations of amino acids in the ATP binding site of 5,10-methenyltetrahydrofolate synthetase (2012), Protein J., 31, 519-528.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.3.2 expressed in Escherichia coli BL21(DE3)/pSJS1244 cells Mycoplasmoides pneumoniae

Protein Variants

EC Number Protein Variants Comment Organism
6.3.3.2 D124A inactive Mycoplasmoides pneumoniae
6.3.3.2 D151A the mutant shows increased turnover number compared to the wild type enzyme Mycoplasmoides pneumoniae
6.3.3.2 D154A inactive Mycoplasmoides pneumoniae
6.3.3.2 D81A the mutant shows reduced turnover number compared to the wild type enzyme Mycoplasmoides pneumoniae
6.3.3.2 K120A the mutant shows increased turnover number compared to the wild type enzyme Mycoplasmoides pneumoniae
6.3.3.2 K3A the mutant shows strongly reduced turnover number compared to the wild type enzyme Mycoplasmoides pneumoniae
6.3.3.2 K3D/D151K the mutant shows strongly reduced turnover number compared to the wild type enzyme Mycoplasmoides pneumoniae
6.3.3.2 K3D/D154 K inactive Mycoplasmoides pneumoniae
6.3.3.2 R125A the mutant shows reduced turnover number compared to the wild type enzyme Mycoplasmoides pneumoniae
6.3.3.2 R7A the mutant shows severely reduced turnover number compared to the wild type enzyme Mycoplasmoides pneumoniae
6.3.3.2 W153A the mutant shows reduced turnover number compared to the wild type enzyme Mycoplasmoides pneumoniae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 130
-
 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 Mycoplasmoides pneumoniae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.3.2 ATP + 5-formyltetrahydrofolate Mycoplasmoides pneumoniae
-
 ADP + phosphate + 5,10-methenyltetrahydrofolate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.3.2 Mycoplasmoides pneumoniae
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.3.2 Talon metal affinity column chromatography Mycoplasmoides pneumoniae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.3.2 ATP + 5-formyltetrahydrofolate
-
 Mycoplasmoides pneumoniae ADP + phosphate + 5,10-methenyltetrahydrofolate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.3.3.2 5,10-Methenyltetrahydrofolate synthetase
-
 Mycoplasmoides pneumoniae
6.3.3.2 MTHFS
-
 Mycoplasmoides pneumoniae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae
6.3.3.2 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 Mycoplasmoides pneumoniae

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.3.2 ATP
-
 Mycoplasmoides pneumoniae