Information on EC 1.5.1.5 - methylenetetrahydrofolate dehydrogenase (NADP+)

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

EC NUMBER
COMMENTARY hide
1.5.1.5
-
RECOMMENDED NAME
GeneOntology No.
methylenetetrahydrofolate dehydrogenase (NADP+)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
5,10-methylenetetrahydrofolate + NADP+ = 5,10-methenyltetrahydrofolate + NADPH + H+
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Carbon fixation pathways in prokaryotes
-
-
folate transformations I
-
-
folate transformations II
-
-
formate assimilation into 5,10-methylenetetrahydrofolate
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
N10-formyl-tetrahydrofolate biosynthesis
-
-
One carbon pool by folate
-
-
purine nucleobases degradation II (anaerobic)
-
-
reductive acetyl coenzyme A pathway I (homoacetogenic bacteria)
-
-
purine metabolism
-
-
reductive acetyl coenzyme A pathway
-
-
SYSTEMATIC NAME
IUBMB Comments
5,10-methylenetetrahydrofolate:NADP+ oxidoreductase
In eukaryotes, occurs as a trifunctional enzyme also having methenyltetrahydrofolate cyclohydrolase (EC 3.5.4.9) and formate---tetrahydrofolate ligase (EC 6.3.4.3) activity. In some prokaryotes occurs as a bifunctional enzyme also having methenyltetrahydrofolate cyclohydrolase activity (EC 3.5.4.9).
CAS REGISTRY NUMBER
COMMENTARY hide
9029-14-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
var.durans Ak, an amethopterin-resistent mutant
-
-
Manually annotated by BRENDA team
strain B, ATCC 11303
-
-
Manually annotated by BRENDA team
male white leghorns, 6-9 months
-
-
Manually annotated by BRENDA team
no activity in Methylobacterium extorquens
-
-
-
Manually annotated by BRENDA team
L. cv. Homesteader
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
5,10-methylene tetrahydrofolate + NAD+
5,10-methenyl tetrahydrofolate + NADH + H+
show the reaction diagram
-
bifunctional enzyme exhibits dehydrogenase and cyclohydrogenase activities
-
-
r
5,10-methylene tetrahydrofolate + NADP+
5,10-methenyl tetrahydrofolate + NADPH
show the reaction diagram
-
trifunctional enzyme exhibits synthetase, dehydrogenase and cyclohydrolase activities
-
-
r
5,10-methylene tetrahydrofolate + NADP+
5,10-methenyl tetrahydrofolate + NADPH + H+
show the reaction diagram
5,10-methylene tetrahydrofolate + NADP+
5,10-methenyltetrahydrofolate + NADPH
show the reaction diagram
dehydrogenase activity of MTHFD1
-
-
r
5,10-methylene-tetrahydrofolate + NAD+
5,10-methenyl-tetrahydrofolate + NADH
show the reaction diagram
-
combined cofactors NAD+, Mg2+, and phosphate mimic the binding of NADP+ in NADP(+)-dependent dehydrogenase
50-60% channeled to the cyclohydrolase to form formyltetrahydrofolate
-
r
5,10-methylene-tetrahydrofolate + NADP+
5,10-methenyl-tetrahydrofolate + NADPH + H+
show the reaction diagram
5,10-methylenetetrahydrofolate + NADP+
5,10-methenyltetrahydrofolate + NADPH + H+
show the reaction diagram
5,10-methylenetetrahydropteroate + NADP+
5,10-methenyltetrahydropteroate + NADPH
show the reaction diagram
-
-
-
-
?
5,10-methylenetetrahydropteroyltriglutamate + NADP+
5,10-methenyltetrahydropteroyltriglutamate + NADPH
show the reaction diagram
DL-tetrahydrofolic acid + NADP+
?
show the reaction diagram
-
-
-
-
?
DL-tetrahydropteroyl-L-aspartate + NADP+
?
show the reaction diagram
-
-
-
-
?
formaldehyde + NADP+
?
show the reaction diagram
-
-
-
-
?
tetrahydropteroylglutamic acid + NADP+
?
show the reaction diagram
-
-
-
-
?
tetrahydropteroylpentaglutamate + NADP+
?
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
5,10-methylene tetrahydrofolate + NAD+
5,10-methenyl tetrahydrofolate + NADH + H+
show the reaction diagram
-
bifunctional enzyme exhibits dehydrogenase and cyclohydrogenase activities
-
-
r
5,10-methylene tetrahydrofolate + NADP+
5,10-methenyl tetrahydrofolate + NADPH
show the reaction diagram
-
trifunctional enzyme exhibits synthetase, dehydrogenase and cyclohydrolase activities
-
-
r
5,10-methylene tetrahydrofolate + NADP+
5,10-methenyl tetrahydrofolate + NADPH + H+
show the reaction diagram
5,10-methylene tetrahydrofolate + NADP+
5,10-methenyltetrahydrofolate + NADPH
show the reaction diagram
P11586
dehydrogenase activity of MTHFD1
-
-
r
5,10-methylene-tetrahydrofolate + NAD+
5,10-methenyl-tetrahydrofolate + NADH
show the reaction diagram
-
combined cofactors NAD+, Mg2+, and phosphate mimic the binding of NADP+ in NADP(+)-dependent dehydrogenase
50-60% channeled to the cyclohydrolase to form formyltetrahydrofolate
-
r
5,10-methylene-tetrahydrofolate + NADP+
5,10-methenyl-tetrahydrofolate + NADPH + H+
show the reaction diagram
5,10-methylenetetrahydrofolate + NADP+
5,10-methenyltetrahydrofolate + NADPH + H+
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
inorganic phosphate
-
required for activity, competitive inhibitor of NADP
Mn2+
-
required for activity when Mg2+ is absent
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioate
-
-
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
-
5,10-formyltetrahydrofolate substrate analogue inhibits dehydrogenase activity
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
-
competitive substrate analogue
-
2,2'-dipyridyl
-
-
2,4-diamino-6-(3,4-dichlorophenoxy)-quinazoline
-
-
2,4-diamino-6-benzyl-5-(3-phenylpropyl)-pyrimidine
-
-
Borate
-
inhibition reversible by potassium
dihydropteroylmonoglutamate
-
competitive inhibition
dihydropteroylpentaglutamate
-
competitive inhibition
folic acid
formaldehyde
guanine
-
48% inhibition
iodoacetate
-
-
Iodosobenzoate
-
-
LY354899
i.e. 5,6,7,8-tetrahydro N5,N10-carbonylfolic acid
methotrexate
-
-
N-ethylmaleimide
N10-formyltetrahydrofolate
-
-
NADPH
o-phenanthroline
-
-
p-chloromercuribenzoate
phenacyl bromide
-
-
phosphate
-
competitive inhibitor against NADP+
potassium phosphate
-
effect on NADP+-dependent dehydrogenase activity
potassium sulfate
-
effect on NADP+-dependent dehydrogenase activity
pteroylglutamate
-
competitive inhibition
pteroylglutamates
additional information
-
no inhibition by pteridine analogues, such as methotrexate, or by hydrophobic diamino-quinazoline, -pyrimidine or -pteridine analogues, such as 2,4-diamino-6-(3,4-dichlorophenoxy)-quinazoline, or 2,4-diamino-6-benzyl-5-(3-phenylpropyl)-pyrimidine, or 2,4-diamino-6,7-diisopropylpteridine
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,3-Dimercaptopropanol
-
the most effective thiol
2-mercaptoethanol
-
-
formaldehyde
-
maximal activity at 0.3 mM
phosphate
-
effect on the NAD+-dependent dehydrogenase activity
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.12
5,10-methylene-tetrahydrofolate
-
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27C; recombinant DHCH1
0.026 - 0.17
5,10-methylenetetrahydrofolate
1.6
5,10-Methylenetetrahydropteroate
-
-
0.0088
5,10-Methylenetetrahydropteroyltriglutamate
-
-
0.21
DL-tetrahydrofolic acid
-
-
0.79
formaldehyde
-
-
0.0011 - 0.039
methylenetetrahydrofolate
0.507 - 4.76
NAD+
0.0004 - 0.69
NADP+
0.0077
tetrahydropteroylglutamic acid
-
-
0.021
tetrahydropteroylmonoglutamate
-
-
0.025
Tetrahydropteroylpentaglutamate
-
-
additional information
NADP+
-
mutation of Arg173 causes a 500fold increase in the Km value for NADP+, while mutation of Ser197 causes a 20fold increase
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
14
5,10-methylene tetrahydrofolate
Leishmania major
-
recombinant DHCH1, 0.25 mM substrate
14
5,10-methylene-tetrahydrofolate
Leishmania major
-
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27C
3.2 - 9.4
methylenetetrahydrofolate
136
NAD+
Photobacterium phosphoreum
-
-
3.8 - 113
NADP+
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
120
5,10-methylene-tetrahydrofolate
Leishmania major
-
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27C
13829
260
NADP+
Leishmania major
-
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 30 mM 2-mercaptoethanol, 27C, with 250 microM 5,10-methylene-tetrahydrofolate
10
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000105
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioate
-
-
0.000105
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
-
pH 7.3, 27C, Leishmania wild-type growth is inhibited with an EC50 of 1.1 microM, DHCH1 or formate-tetrahydrofolate ligase overexpressing (FTL) Leishmania transfectants show 4-5fold or 3fold resistance, respectively
-
0.035
dihydropteroylmonoglutamate
-
varying concentration of tetrahydropteroylmonoglutamate
0.003
dihydropteroylpentaglutamate
-
varying concentration of tetrahydropteroylmonoglutamate
0.000105
LY354899
pH and temperature not specified in the publication
0.07
N10-formyltetrahydrofolate
-
-
0.61
pteroylglutamate
-
varying concentration of tetrahydropteroylmonoglutamate
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0011 - 0.0051
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0011 - 0.0051
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
-
0.00068 - 0.00076
2,4-diamino-6-(3,4-dichlorophenoxy)-quinazoline
0.0004 - 0.00044
2,4-diamino-6-benzyl-5-(3-phenylpropyl)-pyrimidine
0.000036 - 0.000041
methotrexate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.009
-
mitochondria
0.011
-
mitochondria
0.028
-
mitochondria
0.029
-
mitochondria
0.038
-
soluble
0.05
-
soluble
0.092
-
soluble
0.164
-
soluble
2.4 - 6.9
-
bifunctional NAD+-dependent enzyme, in various cell lines
2.7 - 12.9
-
bifunctional NAD+-dependent enzyme, in various cell lines
6.53
-
-
8 - 17.9
-
trifuncional NADP+-dependent enzyme, in various cell lines
8.1
-
trifunctional enzyme, human liver
12.7 - 42
-
trifunctional NADP+-dependent enzyme, in various cell lines
13.2
dehydrogenase activity of the wild type enzyme
13.9
dehydrogenase activity of the mutant R653Q
22
-
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27C, 6.3 microM/min/mg activity of the methenyltetrahydrofolate cyclohydrolase; dehydrogenase activity is measured by following the conversion of 5,10-methylenetetrahydrofolate to 5,10-methenyltetrahydrofolate
32.5
-
-
97.4
-
recombinant bifunctional enzyme
310
-
assays at 37C
720
-
at 60C growth temperature
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 7
-
-
6.8 - 8
-
half-maximal activities at pH 5 and 9.8
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
27
-
assay at
30
-
enzyme assay
39.5
-
potassium phosphate buffer, pH 7.8
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Campylobacter jejuni subsp. jejuni serotype O:2 (strain ATCC 700819 / NCTC 11168)
Francisella tularensis subsp. tularensis (strain SCHU S4 / Schu 4)
Methylobacterium extorquens (strain ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1)
Methylobacterium extorquens (strain ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Thermoplasma acidophilum (strain ATCC 25905 / DSM 1728 / JCM 9062 / NBRC 15155 / AMRC-C165)
Thermoplasma acidophilum (strain ATCC 25905 / DSM 1728 / JCM 9062 / NBRC 15155 / AMRC-C165)
Trypanosoma brucei brucei (strain 927/4 GUTat10.1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30000
-
2 * 30000, SDS-PAGE
30600
-
SDS-PAGE
31900
2 * 31900, calculated from amino acid sequence
33000
-
x * 33000, fragment has methylenetetrahydrofolate dehadrogenase and methenyltetrahydrofolate cyclohydrase activity, proteolysis, SDS-PAGE
34000
-
MTHFD2 evolves from a trifunctional dehydrogenase-cyclohydrolase-synthetase precursor with the loss of synthetase function
38500
-
? * 38500, probably not a heterodimer, SDS-PAGE
50000
-
equilibrium ultracentrifugation
55000
-
methylenetetrahydrofolate dehydrogenase and methenyltetrahydrofolate cyclohydrolase combined, equilibrium ultracentrifugation
57000
-
1 * 57000, bifunctional protein: methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase, SDS-PAGE
58000
-
methylenetetrahydrofolate dehydrogenase-methyltetrahydrofolate cyclohydrolase protein, gel filtration
60000
-
methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase protein, gel filtration
65000
gel filtration
70000
-
sucrose density gradient centrifugation
100000
101000
-
trifunctional enzyme: 5,10-methylenetetrahydrofolate dehydrogenase, 5,10-methenyltetrahydrofolate cyclohydrolase and 10-formyltetrahydrofolate synthetase, transcription-translation of a cDNA clone
104500
-
2 * 104500, SDS-PAGE
108000
-
2 * 108000, 1 * 218000 intramolecular cross-linked with dimethylsuberimidate, SDS-PAGE
150000
-
methylenetetrahydrofolate dehydrogenase, formyltetrahydrofolate synthetase and methenyltetrahydrofolate cyclohydrolase combined, gel filtration
201000
218000
-
2 * 108000, 1 * 218000 intramolecular cross-linked with dimethylsuberimidate, SDS-PAGE
226000
-
methylenetetrahydrofolate dehydrogenase, formyltetrahydrofolate synthetase and methenyltetrahydrofolate cyclohydrolase combined, gel filtration
228000
-
methylenetetrahydrofolate dehydrogenase, formyltetrahydrofolate synthetase and methenyltetrahydrofolate cyclohydrolase combined, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, using 20% (w/v) PEG 3350 and 0.2 M ammonium acetate
hanging drop vapour diffusion method, in the presence of its cofactor NADP+, at 22C using polyethylene glycol 4000 as a precipitant
-
native form and as a NADP complex, hanging drop vapor diffusion method, at 22C in 18% PEG 4000, 400 mM NaCl, and 100 mM Tris-HCl (pH 8.0)
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37 - 50
-
30 min
37
-
30 min stable, destroyed in 3 min at 60C
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
extremely labile, during purification protection by NADP+
-
freezing inactivates
-
frozen stable for 12 days, loss of activity upon repeated freezing and thawing
-
highly instable, partial protection against inactivation by thiol compounds, glycerol, serum albumin and EDTA, freezing inactivates purified enzyme
-
highly unstable, can be preserved as suspension in ammonium sulfate at 50% saturation
-
protected by NADP+
-
relatively instable after partial purification
-
relatively labile, stabilized with phosphate, glycerol and NADP+
-
stable for 1 week, stored in 20% glycerol, 0.1 M potassium phosphate, pH 7.3
-
upon daily freezing and thawing activity loss of 30% in 2 weeks
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-17C, partially purified enzyme after second ammonium sulphate fractionation stable for 2 weeks, 1 mM mercaptoethanol,
-
-20C, 0.1% bovine plasma albumin
-
-20C, 1 month, residual activity of 50%
-
-20C, stable for at least 2 months at diluted protein concentration
-
-20C, stable fot at least 2 months
-
0-4C, after 1 month retains 75% of the methylenetetrahydrofolate dehydrogenase activity
-
10% glycerol required to prevent loss of activity during storage
-
2-4C, 10% loss of activity in 4 days
-
4C, in the presence of 25% glycerol, 1 month, residual activity of 85%
-
4C, stable for at least 2 months
-
4C, stable for at least a month
-
5C, stable for several weeks in ammonium sulfate at 50% saturation
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
bifunctional methylenetetrahydrofolate dehydrogenase-cyclohydrolase
by anion-exchange chromatography
co-purification of 5,10-methylenetetrahydrofolate dehydrogenase and 5,10-methenyltetrahydrofolate cyclohydrolase
-
complex of N5,N10-methylenetetrahydrofolate dehydrogenase and N5,N10-methenyltetrahydrofolate cyclohydrolase
-
formyltetrahydrofolate synthetase, methenyltetrahydrofolate cyclohydrolase and methylenetetrahydrofolate dehydrogenase combined
-
heat treatment, Ni-affinity column chromatography, and Superdex-200 gel filtration
His-Trap affinity column chromatography
-
HisTrap column chromatography and Superdex 200 gel filtration
in vitro transcription-translation of cDNA clone, trifunctional enzyme: 5,10-methylenetetrahydrofolate dehydrogenase, 5,10-methenyltetrahydrofolate cyclohydrase and 10-formyltetrahydrofolate synthetase
-
metal affinity chromatography; recombinant DHCH1 protein is purified by metal-affinity chromatography
-
methylenetetrahydrofolate dehydrogenase and methenyltetrahydrofolate cyclohydrolase combined
-
partial
trifunctional enzyme: methylenetetrahydrofolate dehydrogenase, methenyltetrahydrofolate cyclohydrase, formyltetrahydrofolate synthetase
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
deletion of the MIS1 gene has little effect
-
expressed in Escherichia coli
expressed in Escherichia coli BL21(DE3) cells
expressed in Saccharomyces cerevisiae strain MWY4.4 and in CHO cells
expression in Escherichia coli
-
hexa-His tagged DHCH1 is expressed in Escherichia coli, production of a genetic knock-out; PCR-amplification, expressed in Escherichia coli, overexpression and null-mutant production
-
in vitro transcription-translation of a cDNA clone
-
null mutation is embryonic lethal at about 12 days gestation, knockout of enzyme establishes the important role of formate production during embryogenesis
-
the wild type and R653Q mutant pBKeDCS constructs are transformed into Escherichia coli BL21 DE3 to express the full-length protein
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D133E
-
mutant retains no dehydrogenase activity; no dehydrogenase activity retained, Mg2+ binding site
K56Q/Q100K
-
retaines two-third of dehydrogenase activity but no cyclohydrolase activity; site-directed mutagenesis shows that the double mutant has no cyclohydrolase activity but retains two-thirds of the normal dehydrogenase activity
R653Q
mutant enzyme has normal substrate affinity but a 36% reduction in half-life at 42C
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
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