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Literature summary for 6.3.3.2 extracted from

  • Hancock, A.N.; Coleman, R.S.; Johnson, R.T.; Sarisky, C.A.; Johann, T.W.
    Investigations of the roles of arginine 115 and lysine 120 in the active site of 5,10-methenyltetrahydrofolate synthetase from Mycoplasma pneumoniae (2008), Protein J., 27, 303-308.
    View publication on PubMed
Search for more literature for 6.3.3.2

Cloned(Commentary)

Cloned (Comment) Organism
wild type and mutant enzymes are expressed in Escherichia coli strain BL21(DE3)/pSJS1244 Mycoplasma pneumoniae

Protein Variants

Protein Variants Comment Organism
K120A the mutant shows increased Km and turnover number values for ATP and 5-formyltetrahydrofolate compared to the wild type enzyme Mycoplasma pneumoniae
R115A the mutant shows no activity Mycoplasma pneumoniae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0025
-
 5-formyltetrahydrofolate wild type enzyme, in 200 mM MES pH 6.0, with 30 mM NaCl, 10 mM MgCl2, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol, at 37°C Mycoplasma pneumoniae
0.0071
-
 5-formyltetrahydrofolate mutant enzyme K120A, in 200 mM MES pH 6.0, with 30 mM NaCl, 10 mM MgCl2, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol, at 37°C Mycoplasma pneumoniae
0.076
-
 ATP wild type enzyme, in 200 mM MES pH 6.0, with 30 mM NaCl, 10 mM MgCl2, 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, with 30 mM NaCl, 10 mM MgCl2, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol, at 37°C Mycoplasma pneumoniae

Organism

Organism UniProt Comment Textmining
Mycoplasma pneumoniae P75430
-
-

Purification (Commentary)

Purification (Comment) Organism
Talon metal affinity resin chromatography and Sephadex G-25 gel filtration Mycoplasma pneumoniae

Substrates and Products (Substrate)

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

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
0.79
-
 5-formyltetrahydrofolate wild type enzyme, in 200 mM MES pH 6.0, with 30 mM NaCl, 10 mM MgCl2, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol, at 37°C Mycoplasma pneumoniae
1
-
 ATP wild type enzyme, in 200 mM MES pH 6.0, with 30 mM NaCl, 10 mM MgCl2, 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 K120A, in 200 mM MES pH 6.0, with 30 mM NaCl, 10 mM MgCl2, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol, at 37°C Mycoplasma pneumoniae
1.7
-
 ATP mutant enzyme K120A, in 200 mM MES pH 6.0, with 30 mM NaCl, 10 mM MgCl2, 0.1% (v/v) Triton X-100, and 0.01% (v/v) 2-mercaptoethanol, at 37°C Mycoplasma pneumoniae