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Information on EC 6.3.3.2 - 5-formyltetrahydrofolate cyclo-ligase and Organism(s) Mycoplasma pneumoniae and UniProt Accession P75430
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6.3.3.2 5-formyltetrahydrofolate cyclo-ligaseSpecify your search results
Word Map
- 6.3.3.2
- mthfr
- methylenetetrahydrofolate
-
one-carbon
- homocysteine
-
folate-dependent
-
slc19a1
- 10-formyltetrahydrofolate
-
trifunctional
-
cyclohydrolase
- 5,10-methylenetetrahydrofolate
-
remethylation
-
folate-mediated
-
folate-related
-
1-carbon
-
megaloblastic
-
periconceptional
-
folate-metabolizing
- analysis
- pharmacology
- medicine
The taxonomic range for the selected organisms is: Mycoplasma pneumoniae
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
mthfd1, mthfs, methenyltetrahydrofolate synthetase, 5,10-methenyltetrahydrofolate synthetase, 5-fcl, 5-formyltetrahydrofolate cyclo-ligase, fau1p, 5-cho-thf cycloligase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
5,10-methenyl-tetrahydrofolate synthetase
-
-
-
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5,10-Methenyltetrahydrofolate synthetase
5,10-Methenyltetrahydropteroylglutamate synthetase
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-
-
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5-Formyltetrahydrofolate cyclodehydrase
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-
-
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CH+-THF synthetase
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-
-
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Formyltetrahydrofolic cyclodehydrase
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-
-
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Methenyl THFS
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-
-
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Methenyl-THF synthetase
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-
-
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Methenyltetrahydrofolate synthetase
MTHFS
N5-Formyltetrahydrofolic acid cyclodehydrase
-
-
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Synthetase, methenyltetrahydrofolate
-
-
-
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additional information
-
the enzyme belongs to the cycloligase protein family
5,10-Methenyltetrahydrofolate synthetase
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-
-
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Methenyltetrahydrofolate synthetase
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-
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MTHFS
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-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + 5-formyltetrahydrofolate = ADP + phosphate + 5,10-methenyltetrahydrofolate
active site and substrate binding site structure
-
ATP + 5-formyltetrahydrofolate = ADP + phosphate + 5,10-methenyltetrahydrofolate
active site and substrate binding site structure, binding and catalytic mechanism
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
heteroatomic ring closure
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
5-formyltetrahydrofolate cyclo-ligase (ADP-forming)
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CAS REGISTRY NUMBER
COMMENTARY
37318-64-2
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + 5-formyltetrahydrofolate
ADP + 5,10-methenyltetrahydrofolate + phosphate
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + 5,10-methenyltetrahydrofolate + phosphate
-
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methenyltetrahydrofolate
-
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methenyltetrahydrofolate
-
ATP in form of MgATP2-
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + 5-formyltetrahydrofolate
ADP + 5,10-methenyltetrahydrofolate + phosphate
-
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methenyltetrahydrofolate
-
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
no activity with nucleoside 5'-mono- or 5'-diphosphates
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
-
dependent on, most effective divalent cation, required for MgATP2- complex formation, binding structure
additional information
-
the enzyme is dependent on divalent cation, Mg2+ is preferred
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
steady-state kinetics, recombinant enzyme
-
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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
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
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
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
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
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
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
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
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
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
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
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
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
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
-
2 * 22000, recombinant enzyme, SDS-PAGE, the recombinant enyme exists as a mixture of monomers and dimers
monomer
-
1 * 22000, recombinant enzyme, SDS-PAGE, the recombinant enyme exists as a mixture of monomers and dimers
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant enzyme complexed with ADP, phosphate, and 5-formyltetrahydrofolate, hanging drop vapour diffusion method, 20°C, 30 mg/ml protein with 20% PEG 4000, 5 mM ATP, and 10 mM 5-formylTHF, flash freezing of all crystals in 20% PEG 4000 and 30% ethylene glycol, X-ray diffraction structure determination and analysis at 2.5 A resolution, crystallization of the enzyme with ATP analogues ATP-gamma-S, AMP-PNP, or AMP-PCP is not successful
-
purified recombinant His-tagged wild-type and selenomethionine-labeled enzyme, hanging drop vapour diffusion method, 30 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 0.1 M NaCl, 1 mM EDTA, 1 mM DTT, at room temperature, 30 mg/ml selenomethionine-labeled enzyme in 0.2 M ammonium sulfate, 0.1 M sodium acetate, 21.25% PEG 4000, 15% PEG 400, pH 4.6, at 20°C, flash freezing of all crystals in a solution containing 0.17 M ammonium sulfate, 0.085 M sodium acetate, 21.25% PEG 4000, 15% PEG 400, pH 4.6, X-ray diffraction structure determination and analysis at 2.2 A resolution
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K120A
the mutant shows increased Km and turnover number values for ATP and 5-formyltetrahydrofolate compared to the wild type enzyme
D151A
-
the mutant shows increased turnover number compared to the wild type enzyme
D81A
-
the mutant shows reduced turnover number compared to the wild type enzyme
K120A
-
the mutant shows increased turnover number compared to the wild type enzyme
K3A
-
the mutant shows strongly reduced turnover number compared to the wild type enzyme
K3D/D151K
-
the mutant shows strongly reduced turnover number compared to the wild type enzyme
R125A
-
the mutant shows reduced turnover number compared to the wild type enzyme
R7A
-
the mutant shows severely reduced turnover number compared to the wild type enzyme
W153A
-
the mutant shows reduced turnover number compared to the wild type enzyme
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Talon metal affinity resin chromatography and Sephadex G-25 gel filtration
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography to homogeneity
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography to over 95% homogeneity
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
wild type and mutant enzymes are expressed in Escherichia coli strain BL21(DE3)/pSJS1244
DNA sequence determination and analysis, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
-
overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Chen, S.; Shin, D.H.; Pufan, R.; Kim, R.; Kim, S.H.
Crystal structure of methenyltetrahydrofolate synthetase from Mycoplasma pneumoniae (GI: 13508087) at 2.2.ANG. resolution
Proteins
56
839-843
2004
Mycoplasma pneumoniae
Chen, S.; Yakunin, A.F.; Proudfoot, M.; Kim, R.; Kim, S.H.
Structural and functional characterization of a 5,10-methenyltetrahydrofolate synthetase from Mycoplasma pneumoniae (GI: 13508087)
Proteins
61
433-443
2005
Mycoplasma pneumoniae
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
Protein J.
27
303-308
2008
Mycoplasma pneumoniae (P75430), Mycoplasma pneumoniae
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