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Information on EC 6.3.3.2 - 5-formyltetrahydrofolate cyclo-ligase and Organism(s) Homo sapiens and UniProt Accession P49914
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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: Homo sapiens
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
-
-
-
-
5,10-Methenyltetrahydrofolate synthetase
5,10-Methenyltetrahydropteroylglutamate synthetase
-
-
-
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5-Formyltetrahydrofolate cyclodehydrase
-
-
-
-
CH+-THF synthetase
-
-
-
-
Formyltetrahydrofolic cyclodehydrase
-
-
-
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Methenyl THFS
-
-
-
-
Methenyl-THF synthetase
-
-
-
-
Methenyltetrahydrofolate synthetase
MTHFS
N5-Formyltetrahydrofolic acid cyclodehydrase
-
-
-
-
Synthetase, methenyltetrahydrofolate
-
-
-
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5,10-Methenyltetrahydrofolate synthetase
-
-
-
-
Methenyltetrahydrofolate synthetase
-
-
-
-
MTHFS
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + 5-formyltetrahydrofolate = ADP + phosphate + 5,10-methenyltetrahydrofolate
sequential mechanism
-
ATP + 5-formyltetrahydrofolate = ADP + phosphate + 5,10-methenyltetrahydrofolate
NMR analysis of the catalytic mechanism
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
heteroatomic ring closure
heteroatomic ring closure
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
5-formyltetrahydrofolate cyclo-ligase (ADP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY
37318-64-2
-
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 + (6R,S)-5-formyltetrahydrofolate
ADP + phosphate + 5,10-methenyltetrahydrofolate
-
-
-
-
?
additional information
?
-
-
increased MTHFS expression affects the efficacy of the glycinamide ribonucleotide formyltransferase inhibitor LY309887, overview. SH-SY5Y neuroblastoma with increased MTHFS expression display a 4fold resistance to the GARFT inhibitor LY309887, but do not exhibit resistance to the thymidylate synthase inhibitor Pemetrexed.
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methenyltetrahydrofolate
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methenyltetrahydrofolate
folate binds in a spherical pocket, substrate binding and active site structure, structure-function analysis, detailed overview
-
-
?
ATP + 5-formyltetrahydrofolate
?
-
obligatory initial metabolic step in the intracellular conversion of 5-formyltetrahydrofolate to other reduced folates
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + 5,10-methenyltetrahydrofolate + phosphate
-
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + 5,10-methenyltetrahydrofolate + phosphate
-
important reaction in one-carbon donor recycling
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + 5,10-methenyltetrahydrofolate + phosphate
-
i.e. 5-FTHF or leucovirin
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methenyltetrahydrofolate
-
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methenyltetrahydrofolate
-
enzyme prefers the polyglutamate forms of the substrate compared to the monoglutamate form
-
ir
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methenyltetrahydrofolate
-
enzyme plays an important role in one-carbon metabolism, overview about folate catabolism
-
ir
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methylenetetrahydrofolate
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methylenetetrahydrofolate
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methylenetetrahydrofolate
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methylenetetrahydrofolate
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methylenetetrahydrofolate
-
no reverse reaction detected
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methylenetetrahydrofolate
-
(6R,S)-5-formyltetrahydrofolate, (6S)-5-formyltetrahydrofolate
-
?
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 + phosphate + 5,10-methenyltetrahydrofolate
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + 5,10-methenyltetrahydrofolate + phosphate
-
important reaction in one-carbon donor recycling
-
-
?
additional information
?
-
-
increased MTHFS expression affects the efficacy of the glycinamide ribonucleotide formyltransferase inhibitor LY309887, overview. SH-SY5Y neuroblastoma with increased MTHFS expression display a 4fold resistance to the GARFT inhibitor LY309887, but do not exhibit resistance to the thymidylate synthase inhibitor Pemetrexed.
-
-
?
ATP + 5-formyltetrahydrofolate
?
-
obligatory initial metabolic step in the intracellular conversion of 5-formyltetrahydrofolate to other reduced folates
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methenyltetrahydrofolate
-
-
-
-
?
ATP + 5-formyltetrahydrofolate
ADP + phosphate + 5,10-methenyltetrahydrofolate
-
enzyme plays an important role in one-carbon metabolism, overview about folate catabolism
-
ir
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
5-formyltetrahydrofolate polyglutamate
-
induced recombinant enzyme expression in cancer cell cultures, increasing effect with increasing chain length
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00003
[6R,S]-10-formyltetrahydrofolate triglutamate
-
at 37°C in 100 mM MES, pH 6.0
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
activity of recombinantly expressed enzyme in different cell cultures
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
T-lymphoblast cell line CEM and its methotrexate resistant subline, colon tumor cell line
-
T-lymphoblast cell line CEM and its methotrexate resistant subline, colon tumor cell line
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
inhibition of MTHFS in human MCF-7 breast cancer cells arrests the growth of cells
physiological function
MTHFS regulates the flow of carbon through the one-carbon metabolic network, which supplies essential components for the growth and proliferation of cells
metabolism
-
the enzyme is part of the folate-dependent one-carbon metabolism in the cytoplasm, overview
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). MTHFS_HUMAN
203
0
23256
Swiss-Prot
Mitochondrion (Reliability: 2)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25000
27000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant wild-type MTHFS in a binary complex with ADP, with the N5-iminium phosphate reaction intermediate, or as an enzyme-product complex, hanging drop method at 16°C, 600 nl protein solution, with or without 5 mM 5-formyltetrahydrofolate and 5 mM ATP, are equilibrated against 0.05 ml of reservoir solution containing 100 mM HEPES, pH 6.6, 20 mM MgCl2, 20 mM NiCl2, and 20% w/v PEG 3350, 2 days, X-ray diffraction structure determination and analysis
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D154A
site-directed mutagenesis, the mutant shows 0.9% of the wild-type catalytic activity
E61A
site-directed mutagenesis, the mutant shows 8.6% of the wild-type catalytic activity
F55A
site-directed mutagenesis, the mutant shows 53.6% of the wild-type catalytic activity
F85A
site-directed mutagenesis, the mutant shows 72.3% of the wild-type catalytic activity
K10A
site-directed mutagenesis, the mutant shows 7.5% of the wild-type catalytic activity
K150A
site-directed mutagenesis, the mutant shows wild-type catalytic activity
M58A
site-directed mutagenesis, the mutant shows 121.6% of the wild-type catalytic activity
M90A
site-directed mutagenesis, the mutant shows 49.2% of the wild-type catalytic activity
R145A
site-directed mutagenesis, the mutant shows 2.3% of the wild-type catalytic activity
R148A
site-directed mutagenesis, the mutant shows 30.7% of the wild-type catalytic activity
R14A
site-directed mutagenesis, the mutant shows 12.9% of the wild-type catalytic activity
W109A
site-directed mutagenesis, the mutant shows 39.7% of the wild-type catalytic activity
Y152A
site-directed mutagenesis, the mutant shows 24.9% of the wild-type catalytic activity
Y153A
site-directed mutagenesis, the mutant shows 3.2% of the wild-type catalytic activity
Y83A
site-directed mutagenesis, the mutant shows wild-type catalytic activity
additional information
additional information
-
truncation mutant comprising residues 6-202 of the enzyme is fully active and soluble
additional information
splice site mutation (c.1674G>A) identified in a patient suffering MTHFD1 deficiency leads to exon skipping
additional information
-
splice site mutation (c.1674G>A) identified in a patient suffering MTHFD1 deficiency leads to exon skipping
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
non-ionic detergent, 0.1% v/v Tween 20, and 10% v/v glycerol stabilize the enzyme which is extremely labile at 4 C
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, pH 7.0, 0.1% v/v Tween 20, 10% v/v glycerol, stable for several months
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme from Escherichia coli strain BL21 by nickel affinity chromatography
recombinant truncation mutant comprising residues 6-202 from Escherichia coli strain BL21(DE3) by affinity chromatography and substrate competition elution
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
cloning from brain cDNA library, expression of the His-tagged enzyme in Escherichia coli strain BL21
expression in MCF-7 mammary adenocarcinoma cell line and in SH-SY5Y neuroblastoma cell line
-
expression of truncation mutant comprising residues 6-202 in Escherichia coli strain BL21(DE3)
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
medicine
-
the enzyme is important in cancer treatment to rescue cells from high dose levels of the anti-folate methotrexate, and to potentiate the antitumor activity of 5-fluorouracil
medicine
identification of four patients from two different families, due to a missense mutation (c.806C>T, p.Thr296Ile) and a splice site mutation (c.1674G>A) leading to exon skipping,while the other three patients harboured a missense mutation (c.146C>T, p.Ser49Phe) and a premature stop mutation (c.673G>T, p.Glu225*). Patient fibroblasts show severely reduced methionine formation from [14C]-formate, which does not increase in cobalamin supplemented culture medium but is responsive to folic and folinic acid
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Bertrand, R.; MacKenzie, R.E.; Jolivot, J.
Human liver methenyltetrahydrofolate synthetase: improved purification and increased affinity for folate polyglutamate substrates
Biochim. Biophys. Acta
911
154-161
1987
Homo sapiens
Dayan, A.; Bertrand, R.; Beauchemin, M.; Chahla, D.; Mamo, A.; Filion, M.; Skup, D.; Massie, B.; Jolivet, J.
Cloning and characterization of the human 5,10-methenyltetrahydrofolate synthetase-encoding cDNA
Gene
165
307-311
1995
Homo sapiens
Li, M.; Collier, C.; Gritsman, H.; Bertino, J.R.
Activity of 5,10-methenyltetrahydrofolate synthetase in rat tissues and in tumor cell lines
Pteridines
2
75-79
1990
Cricetulus griseus, Homo sapiens, Rattus norvegicus
-
Jolivet, J.
Human 5,10-methenyltetrahydrofolate synthetase
Methods Enzymol.
281
162-170
1997
Homo sapiens
Bertrand, R.; Beauchemin, M.; Dayan, A.; Ouimet, M.; Jolivet, J.
Identification and characterization of human mitochondrial methenyltetrahydrofolate synthetase activity
Biochim. Biophys. Acta
1266
245-2549
1995
Homo sapiens
Anguera, M.C.; Suh, J.R.; Ghandour, H.; Nasrallah, I.M.; Selhub, J.; Stover, P.J.
Methenyltetrahydrofolate synthetase regulates folate turnover and accumulation
J. Biol. Chem.
278
29856-29862
2003
Homo sapiens, Mus musculus
Copeland, E.H.; Ekiel, I.; Cygler, M.
Letter to the editor: 1H, 13C and 15N resonance assignments of human 5,10-methenyltetrahydrofolate synthetase
J. Biomol. NMR
29
547-548
2004
Homo sapiens
Field, M.S.; Szebenyi, D.M.; Stover, P.J.
Regulation of de novo purine biosynthesis by methenyltetrahydrofolate synthetase in neuroblastoma
J. Biol. Chem.
281
4215-4221
2006
Homo sapiens
Field, M.; Anguera, M.; Page, R.; Stover, P.
5,10-Methenyltetrahydrofolate synthetase activity is increased in tumors and modifies the efficacy of antipurine LY309887
Arch. Biochem. Biophys.
481
145-150
2009
Homo sapiens, Mus musculus
Wu, D.; Li, Y.; Song, G.; Cheng, C.; Zhang, R.; Joachimiak, A.; Shaw, N.; Liu, Z.J.
Structural basis for the inhibition of human 5,10-methenyltetrahydrofolate synthetase by N10-substituted folate analogues
Cancer Res.
69
7294-7301
2009
Homo sapiens (P49914), Homo sapiens
Burda, P.; Kuster, A.; Hjalmarson, O.; Suormala, T.; Buerer, C.; Lutz, S.; Roussey, G.; Christa, L.; Asin-Cayuela, J.; Kollberg, G.; Andersson, B.A.; Watkins, D.; Rosenblatt, D.S.; Fowler, B.; Holme, E.; Froese, D.S.; Baumgartner, M.R.
Characterization and review of MTHFD1 deficiency four new patients, cellular delineation and response to folic and folinic acid treatment
J. Inherit. Metab. Dis.
38
863-872
2015
Homo sapiens (P11586), Homo sapiens
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