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Literature summary for 6.3.3.2 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
Search for more literature for 6.3.3.2

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3)/pSJS1244 cells Mycoplasma pneumoniae

Protein Variants

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 [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/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Mycoplasma pneumoniae
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Talon metal affinity column chromatography Mycoplasma pneumoniae

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
5,10-Methenyltetrahydrofolate synthetase
-
 Mycoplasma pneumoniae
MTHFS
-
 Mycoplasma pneumoniae

Turnover Number [1/s]

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

Cofactor Comment Organism Structure
ATP
-
 Mycoplasma pneumoniae