Information on EC 1.5.1.20 - methylenetetrahydrofolate reductase [NAD(P)H]

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
1.5.1.20
-
RECOMMENDED NAME
GeneOntology No.
methylenetetrahydrofolate reductase [NAD(P)H]
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
5-methyltetrahydrofolate + NAD(P)+ = 5,10-methylenetetrahydrofolate + NAD(P)H + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
folate transformations I
-
-
folate transformations II
-
-
N10-formyl-tetrahydrofolate biosynthesis
-
-
reductive acetyl coenzyme A pathway I (homoacetogenic bacteria)
-
-
sulfopterin metabolism
-
-
tetrahydrofolate metabolism
-
-
One carbon pool by folate
-
-
Carbon fixation pathways in prokaryotes
-
-
Metabolic pathways
-
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Microbial metabolism in diverse environments
-
-
SYSTEMATIC NAME
IUBMB Comments
5-methyltetrahydrofolate:NAD(P)+ oxidoreductase
A flavoprotein (FAD). Menadione can also serve as an electron acceptor.
CAS REGISTRY NUMBER
COMMENTARY hide
71822-25-8
-
9028-69-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
ecotype Columbia, expression in Saccharomyces cerevisiae
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
Q7WQX3
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
Q8FZN0
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
guinea pig
-
-
Manually annotated by BRENDA team
Cercocebus sp.
monkey
-
-
Manually annotated by BRENDA team
strain ATCC 23439
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
Q72DD2
UniProt
Manually annotated by BRENDA team
expression in Saccharomyces cerevisiae
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
formerly Mesorhizobium loti
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
two isoenzymes, encoded by the genes MET12 and MET13
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
synonym Sinorhizobium meliloti
UniProt
Manually annotated by BRENDA team
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
-
mutations in the enzyme lead to hyperhomocysteinemia. A C677T polymorphism is associated with an increased risk for the development of cardiovascular disease, Alzheimer's disease, and depression in adults and of neural tube defects in the fetus
metabolism
-
the enzyme plays a key role in folate metabolism and in the homeostasis of homocysteine
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(6R)-5,10-methylenetetrahydrofolate + NAD(P)H + H+
(6S)-5-methyltetrahydrofolate + NAD(P)+
show the reaction diagram
(6R,S)-5,10-methylenetetrahydrofolate + ?
?
show the reaction diagram
5,10-methylene-5,6,7,8-tetrahydropteroylpentaglutamate + reduced acceptor
(+)-5-methyl-5,6,7,8-tetrahydropteroylpentaglutamate + oxidized acceptor
show the reaction diagram
-
equilibrium lies far in favor of 5-methyl-5,6,7,8-tetrahydropteroylpentaglutamate formation, pentaglutamate form binds to the same enzyme site as monoglutamate form
menadione as acceptor for 5,10-methylene-5,6,7,8-tetrahydropteroylpentaglutamate formation
?
5,10-methylenetetrahydrofolate + acceptor
5-methyltetrahydrofolate + reduced acceptor
show the reaction diagram
5,10-methylenetetrahydrofolate + FADH2
5-methyltetrahydrofolate + FAD
show the reaction diagram
5,10-methylenetetrahydrofolate + NAD(P)H
5-methyltetrahydrofolate + NAD(P)+
show the reaction diagram
-
NADH is the preferred cofactor of the plant enzyme
-
-
r
5,10-methylenetetrahydrofolate + NAD(P)H + H+
5-methyltetrahydrofolate + NAD(P)+
show the reaction diagram
5,10-methylenetetrahydrofolate + NAD+
5,10-methenyltetrahydrofolate + NADH + H+
show the reaction diagram
5,10-methylenetetrahydrofolate + NADH
5-methyltetrahydrofolate + NAD+
show the reaction diagram
5,10-methylenetetrahydrofolate + NADP+
5,10-methenyltetrahydrofolate + NADPH + H+
show the reaction diagram
5,10-methylenetetrahydrofolate + NADPH
5-methyltetrahydrofolate + NADP+
show the reaction diagram
5,10-methylenetetrahydrofolate + NADPH
?
show the reaction diagram
5,10-methylenetetrahydrofolate + NADPH + H+
5-methyltetrahydrofolate + NADP+
show the reaction diagram
-
-
-
-
?
5,10-methylenetetrahydrofolate + reduced acceptor
5-methyltetrahydrofolate + oxidized acceptor
show the reaction diagram
5,10-methylenetetrahydrofolate + reduced ferredoxin
5-methyltetrahydrofolate + oxidized ferredoxin
show the reaction diagram
5,10-methylenetetrahydropteroylpoly-L-glutamate + NADP+
5,10-methenyltetrahydropteroylpoly-L-glutamate + NADPH + H+
show the reaction diagram
5-methyltetrahydrofolate + NAD+
5,10-methylenetetrahydrofolate + NADH
show the reaction diagram
5-methyltetrahydrofolate + NADP+
5,10-methylenetetrahydrofolate + NADPH
show the reaction diagram
5-methyltetrahydrofolate + oxidized menadione
5,10-methylenetetrahydrofolate + reduced menadione
show the reaction diagram
5-methyltetrahydropteroylmonoglutamate + 5,10-methylenetetrahydropteroylhexaglutamate
5,10-methylenetetrahydropteroylmonoglutamate + 5-methyltetrahydropteroylhexaglutamate
show the reaction diagram
-
-
-
r
NADH + menadione
NAD+ + reduced menadione
show the reaction diagram
-
-
-
-
-
NADPH + H+ + menadione
NADP+ + menadiol
show the reaction diagram
nitrate + reduced acceptor
nitrite + acceptor
show the reaction diagram
nitrate + reduced benzyl viologen
nitrite + benzyl viologen
show the reaction diagram
nitrate + reduced methyl viologen
nitrite + methyl viologen
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(6R)-5,10-methylenetetrahydrofolate + NAD(P)H + H+
(6S)-5-methyltetrahydrofolate + NAD(P)+
show the reaction diagram
5,10-methylenetetrahydrofolate + FADH2
5-methyltetrahydrofolate + FAD
show the reaction diagram
5,10-methylenetetrahydrofolate + NAD(P)H + H+
5-methyltetrahydrofolate + NAD(P)+
show the reaction diagram
5,10-methylenetetrahydrofolate + NAD+
5,10-methenyltetrahydrofolate + NADH + H+
show the reaction diagram
5,10-methylenetetrahydrofolate + NADH
5-methyltetrahydrofolate + NAD+
show the reaction diagram
5,10-methylenetetrahydrofolate + NADP+
5,10-methenyltetrahydrofolate + NADPH + H+
show the reaction diagram
5,10-methylenetetrahydrofolate + NADPH
5-methyltetrahydrofolate + NADP+
show the reaction diagram
5,10-methylenetetrahydrofolate + NADPH
?
show the reaction diagram
5,10-methylenetetrahydrofolate + NADPH + H+
5-methyltetrahydrofolate + NADP+
show the reaction diagram
-
-
-
-
?
5,10-methylenetetrahydrofolate + reduced acceptor
5-methyltetrahydrofolate + oxidized acceptor
show the reaction diagram
5,10-methylenetetrahydrofolate + reduced ferredoxin
5-methyltetrahydrofolate + oxidized ferredoxin
show the reaction diagram
5-methyltetrahydrofolate + NAD+
5,10-methylenetetrahydrofolate + NADH
show the reaction diagram
-
-
-
-
r
5-methyltetrahydrofolate + NADP+
5,10-methylenetetrahydrofolate + NADPH
show the reaction diagram
-
-
-
-
?
nitrate + reduced acceptor
nitrite + acceptor
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
FADH2
flavin
-
flavin-dependent enzyme
NADP+
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
iron-sulfur centre
-
enzyme contains 15.2 molecules iron and 19.5 molecules acid-labile sulfur as iron-sulfur clusters
Molybdenum
phosphate
Zinc
-
2.3 molecules zinc per enzyme molecule
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(+)-5-methyl-5,6,7,8-tetrahydropteroylpentaglutamate
-
inhibits at high concentrations
2,4-dihydroxyphenylacetic acid
-
at 0.01 M
5,10-methylenetetrahydrofolate
dicoumarol
-
inhibits menadione reductase activity only
diethyldicarbonate
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20 mM NADP+ and 20 mM NAD+ partially protected the enzyme against inactivation whereas 20 mM nicotinamide gives complete protection
dihydrofolate
dihydropteroylhexaglutamate
-
-
dihydropteroylpolyglutamate
-
folylpolyglutamate
-
overview
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LY309887
-
i.e. 6R-2,5-thienyl-5,10-dideazatetrahydrofolate monoglutamate
menadione
-
inhibits enzyme activity in yeast extracts
Mersalyl
-
80% inhibition at 1 mM
methylenetetrahydrofolate
-
-
N-(4-[[(2,4-diaminopteridin-6-yl)methyl](methyl)amino]benzoyl)-gamma-glutamyl-gamma-glutamyl-gamma-glutamyl-gamma-glutamylglutamic acid
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-
N-(4-[[(2,4-diaminopteridin-6-yl)methyl](methyl)amino]benzoyl)-gamma-glutamyl-gamma-glutamylglutamic acid
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-
N-(4-[[(2,4-diaminopteridin-6-yl)methyl](methyl)amino]benzoyl)glutamic acid
-
-
N-([5-[2-(2,4-diaminoquinazolin-6-yl)ethyl]-2,3-dihydrothiophen-2-yl]carbonyl)-4-methylideneglutamic acid
-
-
N-bromosuccinimide
-
inactivates the enzyme by modification of tryptophan
N-[4-[2-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-4-methylideneglutamic acid
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-
N-[4-[2-(2,4-diaminopyrido[3,2-d]pyrimidin-6-yl)ethyl]benzoyl]-4-methylideneglutamic acid
-
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N-[4-[2-(2,4-diaminoquinazolin-6-yl)ethyl]-2-fluorobenzoyl]-4-methylideneglutamic acid
-
-
N-[4-[2-(2,4-diaminoquinazolin-6-yl)ethyl]benzoyl]-4-methylideneglutamic acid
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-
N-[4-[2-(2-amino-4-methylquinazolin-6-yl)ethyl]benzoyl]-4-methylideneglutamic acid
-
-
NADPH
-
100 mM
p-chloromercuribenzoate
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60% inhibition of the methylenetetrahydrofolate reductase activity at 0.04 mM, 83% inhibition of the menadione reductase activity at 0.04 mM
Phenylglyoxal
-
-
phosphate
Polyglutamate analogues
-
-
-
S-adenosyl-L-methionine
S-adenosylmethionine
tetrahydrofolate
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
A23187
-
treatment of cells results in increase in enzyme mRNA and protein
homocysteine
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treatment of cells results in increase in enzyme mRNA and protein
inositol-requiring enzyme-1
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the induction of MTHFR was also observed after overexpression of inositol-requiring enzyme-1
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S-adenosylmethionine
-
stimulates
thapsigargin
-
treatment of cells results in increase in enzyme mRNA and protein
tunicamycin
-
treatment of cells results in increase in enzyme mRNA and protein
Valproic acid
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300 mg/kg treatment increases MTHFR promoter activity 2.5fold and MTHFR mRNA and protein 3.7fold
additional information
-
induction of MTHFR is observed after overexpression of inositol-requiring enzyme IRE1 and is inhibited by a dominant-negative mutant of IRE1. Tranfection of c-Jun and its activators LiCl and sodium valproate increase MTHFR expression, whereas inhibitor of c-Jun SP600125 reduces activation; transfection of c-Jun and two activators of c-Jun (LiCl and sodium valproate) increase MTHFR expression, MTHFR mRNA is up-regulated by endoplasmic reticulum stress
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.003
(+)-5-methyl-5,6,7,8-tetrahydropteroylpentaglutamate
-
-
0.023
(+)-5-methyltetrahydrofolate
-
-
0.18
(6R)-5,10-methylenetetrahydrofolate
at pH 7.0 and 50C
0.0005 - 0.287
5,10-methylenetetrahydrofolate
0.04 - 0.153
5,10-methylenetetrahydropteroylpoly-L-glutamate
-
0.019 - 0.16
5-methyltetrahydrofolate
0.0019
5-methyltetrahydropteroylhexaglutamate
-
-
0.033
5-methyltetrahydropteroylmonoglutamate
-
-
11.1
benzyl viologen
-
-
0.12
DL-5-methyltetrahydrofolate
-
-
0.0004 - 0.187
methylenetetrahydrofolate
0.147
NAD+
pH 8.0, 30C
0.0027 - 0.585
NADH
0.537
NADP+
pH 8.0, 30C
0.0073 - 0.049
NADPH
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.3 - 8.8
5,10-methylenetetrahydropteroylpoly-L-glutamate
-
0.007 - 50
methylenetetrahydrofolate
3.6
NAD+
pH 8.0, 30C
10.4 - 160
NADH
1.1
NADP+
pH 8.0, 30C
26.7 - 160
NADPH
additional information
additional information
-
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
30.95 - 67.69
5,10-methylenetetrahydropteroylpoly-L-glutamate
-
24.49
NAD+
pH 8.0, 30C
2.05
NADP+
pH 8.0, 30C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.04 - 0.051
(+)-5-methyl-5,6,7,8-tetrahydropteroylpentaglutamate
-
-
0.16 - 0.873
5,10-methylenetetrahydrofolate
0.0065
dihydrofolate
-
-
0.000013
dihydropteroylhexaglutamate
-
-
0.061
methylenetetrahydrofolate
-
recombinant wild-type enzyme, pH 7.2, 4C
0.014
NADH
-
recombinant wild-type enzyme, pH 7.2, 4C, the mutant enzymes all show a Km below 0.0035 mM
0.2
S-adenosylmethionine
-
-
additional information
additional information
-
dihydropteroylpolyglutamate inhibitors with different numbers of glutamyl residues
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0007
N-(4-[[(2,4-diaminopteridin-6-yl)methyl](methyl)amino]benzoyl)-gamma-glutamyl-gamma-glutamyl-gamma-glutamyl-gamma-glutamylglutamic acid
Homo sapiens
-
-
0.0022
N-(4-[[(2,4-diaminopteridin-6-yl)methyl](methyl)amino]benzoyl)-gamma-glutamyl-gamma-glutamylglutamic acid
Homo sapiens
-
-
0.033
N-(4-[[(2,4-diaminopteridin-6-yl)methyl](methyl)amino]benzoyl)glutamic acid
Homo sapiens
-
-
0.0018
N-([5-[2-(2,4-diaminoquinazolin-6-yl)ethyl]-2,3-dihydrothiophen-2-yl]carbonyl)-4-methylideneglutamic acid
Homo sapiens
-
-
0.0035
N-[4-[2-(2,4-diaminopyrido[3,2-d]pyrimidin-6-yl)ethyl]benzoyl]-4-methylideneglutamic acid
Homo sapiens
-
-
0.0012
N-[4-[2-(2,4-diaminoquinazolin-6-yl)ethyl]-2-fluorobenzoyl]-4-methylideneglutamic acid
Homo sapiens
-
-
0.0016
N-[4-[2-(2,4-diaminoquinazolin-6-yl)ethyl]benzoyl]-4-methylideneglutamic acid
Homo sapiens
-
-
additional information
N-[4-[2-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-4-methylideneglutamic acid
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0000424
-
liver homogenate
0.000094
-
wild-type, cell extract
0.00022
Cercocebus sp.
-
reverse reaction
0.000231
-
brain homogenate
0.00029
Cercocebus sp.
-
forward reaction
0.00071
-
forward reaction
0.001
-
reverse reaction
0.0017
-
muscle
0.0045
-
brain
0.0048
-
intestine
0.0082
-
kidney
0.0089
-
pancreas
0.0106
-
liver
0.1
-
-
0.367
-
-
0.413
-
-
1.2
-
-
6.92
-
recombinant enzyme
12.4
-
recombinant enzyme
153
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.3 - 6.4
-
formation of 5-methyltetrahydrofolate
6.3 - 6.9
-
-
6.5
-
NADPH-tetrahydrofolate oxidoreductase activity
6.6
-
methyltetrahydrofolate-menadione oxidoreductase activity
6.6 - 6.7
-
methylenetetrahydrofolate reductase reaction
6.8
-
-
7
-
assay at
7.1
-
menadione reductase reaction
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.8 - 8.6
-
about 60% of activity maximum at pH 5.8, about 50% of activity maximum at pH 8.6, menadione reductase activity
6 - 8
-
pH 6: about 70% of activity maximum, pH 8: about 35% of activity maximum with 5-methyl-5,6,7,8-tetrahydropteroylmonoglutamate, about 65% of activity maximum with 5-methyl-5,6,7,8-tetrahydropteroylpentaglutamate
6.2 - 7.3
-
half-maximal activity at pH 6.2 and pH 7.3, methylenetetrahydrofolate reductase reaction
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 45
-
methylenetetrahydrofolate reductase reaction, slow increase of activity between 25C and 45C