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Information on EC 1.5.1.3 - dihydrofolate reductase and Organism(s) Escherichia coli and UniProt Accession P00383
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1.5.1.3 dihydrofolate reductaseIUBMB Comments
The enzyme from animals and some micro-organisms also slowly reduces folate to 5,6,7,8-tetrahydrofolate.
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Word Map
- 1.5.1.3
- methotrexate
- antifolate
- plasmodium
- falciparum
- hamster
- malaria
- pyrimethamine
- ovary
-
antimalarial
- leukemia
- dihydropteroate
- thymidine
- pyrimidine
- carinii
- polyglutamates
- pneumocystis
-
hydride
- chloroquine
-
casey
-
drug-resistant
- tmp
-
ccrf-cem
- vivax
- glycinamide
- toxoplasma
-
uncomplicated
-
folate-dependent
- mthfr
- folylpolyglutamate
- pemetrexed
-
sublines
- leucovorin
- tetrahydrobiopterin
-
polyglutamylation
- gondii
-
nontranscribed
- sulfamethoxazole
- trimethoprim-sulfamethoxazole
- quinazoline
- 5-methyltetrahydrofolate
- sepiapterin
- integrons
- dihydropteridine
- analysis
- medicine
- artesunate
- synthesis
-
triazine
-
folinic
- 5,10-methylenetetrahydrofolate
- bh4
- biotechnology
- biopterins
- drug development
- pterins
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
dhfr, dihydrofolate reductase, thy-1, dhfr-ts, hdhfr, dihydrofolate reductase-thymidylate synthase, ecdhfr, pcdhfr, r67 dhfr, ts-dhfr, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
7,8-dihydrofolate reductase
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-
-
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dehydrogenase, tetrahydrofolate
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-
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DHFR
DHFR type IIIC
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dihydrofolate reductase
dihydrofolate reductase-thymidylate synthase
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dihydrofolate reductase:thymidylate synthase
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dihydrofolic acid reductase
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dihydrofolic reductase
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ecDHFR
folic acid reductase
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folic reductase
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NADPH-dihydrofolate reductase
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pteridine reductase:dihydrofolate reductase
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reductase, dihydrofolate
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tetrahydrofolate dehydrogenase
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thymidylate synthetase-dihydrofolate reductase
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Trimethoprim resistance protein
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DHFR
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DHFR
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
binding of NADPH is accompanied by release of 38 water molecules, while binding of dihydrofolate is accompanied by the net uptake of water
5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
kinetic mechanism
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5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
catalytic mechanism
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5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
rapid random equilibrium mechanism with NADPH and reduced acetylpyridine adenine nucleotide as cofactor
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5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
cofactor binding mechanism, pH-dependence investigation
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5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
inhibitor binding mechanism
-
5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
analysis of higher energy conformational substrates by NMR relaxation dispersion. The maximum hydride transfer and steady-state turnover rates are governed by the dynamics of transitions between ground and excited states of the intermediates. Model of conformational changes during the catalytic cycle
5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
donor carbon at the hydride transfer transition state resembles the reactant state more than the product state, whereas the acceptor carbon is more productlike, a symptom of transition state imbalance
-
5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
kinetic isotope effect analysis, enzyme requires no no donor-acceptor distance fluctuations and thus no gating
-
5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
model for hydrogen transfer includes contributions from quantum mechanical tunnelling coupled with protein motions that actively modulate the tunnelling distance
5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
study on enzyme ternary complex with substrate analogue folates and oxidized NADP+ cofactor using NMR relaxation methods. Conformational exchanges of protein between a ground state with closed conformation of active site loops and an excited state with loops in occluded conformation. Fluctuations include motions of the nicotinamide ring of the cofactor into and out of the active site
5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
the active-site loop dynamics access a closed conformation, and the accompanying closed to occluded rate constant is comparable to the maximum pH-independent hydride transfer rate constant. The closed to occluded conformational transition in the product ternary complex is a prerequisite for progression through the catalytic cycle. The rate of this process places an effective limit on the maximum rate of the hydride transfer step. The dynamics of the C-terminally associated region is pH-dependent, but the dynamics of the active-site loops and cofactor binding cleft are pH-independent
5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+
transfer of a proR hydride from the C4 atom of NADPH to the C6 position of the dihydropterin ring of DHF, mechanism of the proton and hydride transfer reaction, active site residues D27 and Y100 play a synergistic role in facilitating both the proton transfer and subsequent hydride transfer steps. Residue D27 appears to have a greater effect on the overall rate of conversion of DHF to tetrahydrofolate, Y100 plays an important electrostatic role in modulating the pKa of the N5 of DHF to enable the preprotonation of DHF by an active site water molecule. The D27 and Y100 residues function synergistically to provide an active site environment for the solvent-assisted protonation of N5 and to position the reacting substrates, NADPH and DHF, properly
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
redox reaction
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oxidation
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reduction
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PATHWAY SOURCE
PATHWAYS
BRENDA
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-, -, -, -
SYSTEMATIC NAME
IUBMB Comments
5,6,7,8-tetrahydrofolate:NADP+ oxidoreductase
The enzyme from animals and some micro-organisms also slowly reduces folate to 5,6,7,8-tetrahydrofolate.
CAS REGISTRY NUMBER
COMMENTARY
9002-03-3
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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
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
?
7,8-dihydrofolate + acetylpyridine adenine nucleotide, reduced
5,6,7,8-tetrahydrofolate + acetylpyridine adenine nucleotide, oxidized
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-
-
?
7,8-dihydrofolate + NADH + H+
5,6,7,8-tetrahydrofolate + NAD+
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-
-
-
r
dihydrobiopterin + NADPH
? + NADP+
-
10% of the activity with 7,8-dihydrofolate
-
-
?
10-formyl-dihydrofolate + NADPH + H+
10-formyl-tetrahydrofolate + NADP+
-
-
-
?
10-formyl-dihydrofolate + NADPH + H+
10-formyl-tetrahydrofolate + NADP+
the enzyme is involved in the folate recycling pathway
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-
?
7,8-dihydrofolate + NADPH
5,6,7,8-tetrahydrofolate + NADP+
kinetic mechanism
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-
?
7,8-dihydrofolate + NADPH
5,6,7,8-tetrahydrofolate + NADP+
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equilibrium strongly favors tetrahydrofolate production
-
?
7,8-dihydrofolate + NADPH
5,6,7,8-tetrahydrofolate + NADP+
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equilibrium strongly favors tetrahydrofolate production
-
?
7,8-dihydrofolate + NADPH
5,6,7,8-tetrahydrofolate + NADP+
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equilibrium strongly favors tetrahydrofolate production
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r
7,8-dihydrofolate + NADPH
5,6,7,8-tetrahydrofolate + NADP+
-
equilibrium strongly favors tetrahydrofolate production
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r
7,8-dihydrofolate + NADPH
5,6,7,8-tetrahydrofolate + NADP+
-
cannot use NADH as reducing agent
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-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
r
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
DHFR catalyzes hydride transfer from the cofactor NADPH to 7,8-dihydrofolate to produce tetrahydrofolate requiring electrostatic complementarity between the enzyme and the transition state
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-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
the enzyme is involved in the folate recycling pathway
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-
?
additional information
?
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3-acetylpyridine adenine dinucleotide + dihydrofolate: rapid equilibrium random mechanism
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?
additional information
?
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NADPH + dihydrofolate: steady-state random mechanism
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?
additional information
?
-
enzyme activity decreases with increasing pressure. The Km values for dihydrofolate and NADPH are slightly higher at 200 MPa than at atmospheric pressure. The hydride transfer step is insensitive to pressure, while the dissociation constants of substrates and cofactors increase with pressure
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-
?
additional information
?
-
-
enzyme activity decreases with increasing pressure. The Km values for dihydrofolate and NADPH are slightly higher at 200 MPa than at atmospheric pressure. The hydride transfer step is insensitive to pressure, while the dissociation constants of substrates and cofactors increase with pressure
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-
?
additional information
?
-
free energy perturbation with molecular dynamics simulations of dihydrofolate reductase in complex with dihydrofolate. In the Michaelis complex the pKa is modulated by the Met20 loop fluctuations, providing the largest pKa shift in substrates with a tightly closed loop conformation, in the partially closed/open substrates, the pKa is similar to that in the occluded complex. Conducive to the protonation, tightly closing the Met20 loop enhances the interactions of the cofactor and the substrate with the Met20 side chain and aligns the nicotinamide ring of the cofactor coplanar with the pterin ring of the substrate
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-
?
additional information
?
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-
free energy perturbation with molecular dynamics simulations of dihydrofolate reductase in complex with dihydrofolate. In the Michaelis complex the pKa is modulated by the Met20 loop fluctuations, providing the largest pKa shift in substrates with a tightly closed loop conformation, in the partially closed/open substrates, the pKa is similar to that in the occluded complex. Conducive to the protonation, tightly closing the Met20 loop enhances the interactions of the cofactor and the substrate with the Met20 side chain and aligns the nicotinamide ring of the cofactor coplanar with the pterin ring of the substrate
-
-
?
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
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
?
10-formyl-dihydrofolate + NADPH + H+
10-formyl-tetrahydrofolate + NADP+
the enzyme is involved in the folate recycling pathway
-
-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
-
-
-
r
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
the enzyme is involved in the folate recycling pathway
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NADPH
-
-
392208, 392210, 392212, 392214, 392218, 392220, 392224, 392226, 392239, 392240, 392241, 392277, 654970, 655867, 674351, 693567, 705044, 724168, 724390
NADPH
-
cofactor-enzyme interaction study from x-ray structure, structural and topological comparison with dehydrogenases
NADPH
effects of 1,3,5-triazine-2,4-diamine derivatives on the cofactor NADPH binding, overview
NADPH
transient entry of the reduced nicotinamide moiety into the active site pocket causes a steric clash with the pterin ring of the product tetrahydrofolate, facilitating release of the pterin ring from the active site. Transient binding of the ribose moiety causes helix C to shift, thereby opening the pABG cleft and further assisting product release
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(2,5-dimethyl-1,4-phenylene)bis(methylene) bis(N-[amino(imino)methyl](imidothiocarbamate))
50% inhibition at 0.000075 mM
(4-(1-[(5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(methyl)amino]ethyl)phenyl)methanol
-
1,3-phenylenebis(methylene) bis(N-[amino(imino)methyl](imidothiocarbamate))
50% inhibition at 0.000075 mM
1,4-bis-([N-(1-imino-1-guanidino-methyl)]sulfanylmethyl)-3,6-dimethyl-benzene
binding of the inhibitor has both a favorable entropy and enthalpy of binding. Positive binding cooperativity between inhibitor and the cofactor NADPH. Binding of inhibitor to DHFR is 285fold tighter in the presence of the NADPH cofactor than in its absence
1,4-phenylenebis(methylene) bis(N-[amino(imino)methyl](imidothiocarbamate))
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1,6-bis-(4-fluoro-phenyl)-[1,3,5]triazine-2,4-diamine
-
IC50: 0.011 nM, 0.03 mM 7,8-dihydrofolate
1-(3-ethoxyphenyl)-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
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1-(4-chlorophenyl)-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
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1-(4-ethoxyphenyl)-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
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1-(4-nitrophenyl)-3-[4-[4-[(4-nitrophenyl)carbamoylamino]phenoxy]phenyl]urea
NSC80735, complete inhibition at 1 mM
1-([N-(1-imino-guanidino-methyl)]sulfanylmethyl)-3-trifluoromethyl-benzene
binding of the inhibitor has both a favorable entropy and enthalpy of binding. Positive binding cooperativity between inhibitor and the cofactor NADPH. Binding of inhibitor to DHFR in the absence of NADPH is not observed
1-N,4-N-bis(4-aminophenyl)benzene-1,4-dicarboxamide
NSC55152, complete inhibition at 1 mM
1-[3-(aminomethyl)phenyl]-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
-
1-[3-chloro-4-(5-phenylpentyl)phenyl]-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
uncompetitive versus NADPH, competitive versus 7,8-dihydrofolate
1-[4-(aminomethyl)phenyl]-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
-
1-[4-(dimethylamino)phenyl]-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
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1-[4-[4-(2,4-dichlorophenyl)butyl]phenyl]-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
-
2-(2-((5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(1-(naphthalen-1-yl)ethyl)amino)ethoxy)ethanol
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2-(2-((5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)[1-(naphthalen-1-yl)ethyl]amino)ethoxy)ethanol
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2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
-
-
2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-5-[(2-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
-
-
2-amino-5-[(3,4-dichlorophenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-5-[(3,4-dichlorophenyl)thio]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
-
-
2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]-pyrimidin-4(3H)-one
-
-
2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-5-[(3,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
-
-
2-amino-5-[(3-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
-
-
2-amino-5-[(4-bromophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
-
-
2-amino-5-[(4-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-5-[(4-chlorophenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-5-[(4-fluorophenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-6-ethyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d ]pyrimidin-4(3H)-one
-
-
2-amino-6-ethyl-5-[(4-fluorophenyl)sulfanyl]thieno[2,3-d ]pyrimidin-4(3H)-one
-
-
2-amino-6-ethyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-6-methyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-6-methyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-hydroxy-5-nitrobenzyl N-[amino(imino)methyl]imidothiocarbamate
50% inhibition at 0.0151 mM
2-methoxybenzyl N-[amino(imino)methyl]imidothiocarbamate
50% inhibition at 0.0033 mM
2-[(5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(1-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-8-yl]ethyl)amino]ethanol
-
2-[(5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(methyl)amino]-2-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-yl]ethanol
-
3,4-dichlorobenzyl N-[amino(imino)methyl]imidothiocarbamate
50% inhibition at 0.0037 mM
3,5-bis(trifluoromethyl)benzyl N-[amino(imino)methyl]imidothiocarbamate
50% inhibition at 0.0056 mM
3-((5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(1-(naphthalen-1-yl)ethyl)amino)propanoic acid
-
3-((5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)[1-(naphthalen-1-yl)ethyl]amino)propanoic acid
-
3-(trifluoromethyl)benzyl N-[amino(imino)methyl]imidothiocarbamate
50% inhibition at 0.0043 mM
3-heptyl-4-hydroxy-1H-naphthalen-2-one
-
IC50: 0.48 mM, 0.03 mM 7,8-dihydrofolate
3-methoxybenzyl N-[amino(imino)methyl]imidothiocarbamate
50% inhibition at 0.0038 mM
4-((5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(1-(naphthalen-1-yl)ethyl)amino)butanoic acid
-
4-((5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)[1-(naphthalen-1-yl)ethyl]amino)butanoic acid
-
4-(trifluoromethyl)benzyl N-[amino(imino)methyl]imidothiocarbamate
50% inhibition at 0.013 mM
4-methylbenzyl N-[amino(imino)methyl]imidothiocarbamate
50% inhibition at 0.0126 mM
4-[3-(4,6-diamino-2,2-dimethyl-1,3,5-triazin-1(2H)-yl)benzamido]benzene-1-sulfonyl fluoride
contains a fluorosulfonylphenylaminocarbonyl substitution at R2 on 1-phenyl-6,6-dimethyl-1,3,5-triazine-2,4-diamine ring, and shows the unique behavior of tight-binding and average inhibition, uncompetitive versus NADPH, competitive versus 7,8-dihydrofolate
4-[4,6-diamino-2-(4-chlorophenyl)-1,3,5-triazin-1(2H)-yl]benzene-1-sulfonamide
-
4-[4-[3-(4,6-diamino-2,2-dimethyl-1,3,5-triazin-1-yl)phenyl]butyl]benzenesulfonyl fluoride
uncompetitive versus NADPH, competitive versus 7,8-dihydrofolate
4-[6-[4-(4,6-diamino-2,2-dimethyl-1,3,5-triazin-1-yl)phenyl]hexyl]benzenesulfonyl fluoride
uncompetitive versus NADPH, competitive versus 7,8-dihydrofolate
5-(3,4,5-trimethoxy-benzyl)-pyrimidine-2,4-diamine
-
IC50: 18 nM, 0.03 mM 7,8-dihydrofolate
6,6-dimethyl-1-(4-methylphenyl)-1,6-dihydro-1,3,5-triazine-2,4-diamine
-
6,6-dimethyl-1-[3-(4-phenylbutyl)phenyl]-1,6-dihydro-1,3,5-triazine-2,4-diamine
uncompetitive versus NADPH, competitive versus 7,8-dihydrofolate
6,7-dimethyl-5,6,7,8-tetrahydro-quinazoline-2,4-diamine
-
IC50: 790 nM, 0.03 mM 7,8-dihydrofolate
6-(([4-chlorotricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-yl]amino)methyl)pteridine-2,4-diamine
-
6-(4-trifluoromethyl-phenoxy)-quinazoline-2,4-diamine
-
IC50: 660 nM, 0.03 mM 7,8-dihydrofolate
6-([tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-ylamino]methyl)pteridine-2,4-diamine
-
6-([tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-8-ylamino]methyl)pteridine-2,4-diamine
-
6-p-tolyloxy-quinazoline-2,4-diamine
-
IC50: 190 nM, 0.03 mM 7,8-dihydrofolate
6-p-tolylsulfanyl-quinazoline-2,4-diamine
-
IC50: 310 nM, 0.03 mM 7,8-dihydrofolate
6-[(tricyclo[4.3.0.07,9]nona-1,3,5-trien-3-ylamino)methyl]pteridine-2,4-diamine
-
7-methyl-5,6,7,8-tetrahydro-quinazoline-2,4-diamine
-
IC50: 0.011 nM, 0.03 mM 7,8-dihydrofolate
7-[(4-aminophenyl)methyl]-7H-pyrrolo[3,2-f]quinazoline-1,3-diamine
NSC309401, AMPQD, a substrate analogue with the quinazoline-1, 3-diamine group, tight-binding inhibitor, complete inhibition at 1 mM, competitive inhibition of dihydrofolate binding. Potently inhibits the enzyme by competitive displacement of the substrate dihydrofolic acid, uncompetitive inhibition versus NADPH, the inhibitor has a markedly increased affinity for the NADPH-bound form of the enzyme. The mode of binding of the inhibitor to the enzyme-NADPH binary complex conforms to the slow-onset, tight-binding model
7H-pyrrolo[3,2-f]quinazoline-1,3-diamine
PQD, the lack of (4-aminophenyl)-methyl group at position 7 abolishes the slow-onset of inhibition
epigallocatechin gallate
-
bisubstrate inhibitor, binds both on substrate and cofactor site of dihydrofolate reductase. Detailed study on kinetics
N-(4-chloro-2-cyanophenyl)imidodicarbonimidic diamide
-
IC50: 320 nM, 0.03 mM 7,8-dihydrofolate
N-(4-[(2-amino-6-ethyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid
-
-
N-(4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid
-
-
N-(4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d]pyrimidin-5-yl)sulfanyl]benzoyl)-L-glutamic acid
-
binding mode, overview
N-(5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)-N-[1-(2-ethoxynaphthalen-1-yl)ethyl]-b-alanine
-
N7-(1-(naphthalen-1-yl)ethyl)-N7-propylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(2-aminoethyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(2-aminoethyl)-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(2-ethoxyethyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(2-ethoxyethyl)-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(2-propoxyethyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(2-propoxyethyl)-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(3-ethoxypropyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(3-ethoxypropyl)-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(4-aminobutyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(4-aminobutyl)-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(5-aminopentyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(5-aminopentyl)-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(cyclopropylmethyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(cyclopropylmethyl)-N7-[1-(4-methylnaphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(cyclopropylmethyl)-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(prop-2-en-1-yl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-(prop-2-en-1-yl)-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-benzyl-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-benzyl-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-butyl-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-butyl-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-cyclopropyl-N7-(1-(naphthalen-2-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-cyclopropyl-N7-[1-(naphthalen-2-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-ethyl-N7-(1-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-yl]ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-ethyl-N7-(1-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-8-yl]ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-ethyl-N7-(4-methylbenzyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-ethyl-N7-[1-(4-methylnaphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-methyl-N7-(1-phenylethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-methyl-N7-(1-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-yl]ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-methyl-N7-[1-(4-methylnaphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-methyl-N7-[1-(quinolin-4-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-methyl-N7-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-ylmethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-pentyl-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-pentyl-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[1-(1-benzothiophen-3-yl)ethyl]-N7-(cyclopropylmethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[1-(1-benzothiophen-3-yl)ethyl]-N7-ethylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[1-(1-benzothiophen-3-yl)ethyl]-N7-methylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[1-(2-ethoxynaphthalen-1-yl)ethyl]-N7-methylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[1-(2-ethylnaphthalen-1-yl)ethyl]-N7-methylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[1-(4-chloronaphthalen-1-yl)ethyl]-N7-(cyclopropylmethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[1-(naphthalen-1-yl)ethyl]-N7-propylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[2-(4-fluorophenyl)ethyl]-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[2-(4-fluorophenyl)ethyl]-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[2-(cyclohex-2-en-1-yl)ethyl]-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[2-(cyclohex-2-en-1-yl)ethyl]-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[3-(2-methyl-4H-imidazol-4-yl)propyl]-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[3-(2-methyl-4H-imidazol-4-yl)propyl]-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[3-(2-methylpropoxy)propyl]-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
N7-[3-(2-methylpropoxy)propyl]-N7-[1-(naphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
-
organic mercurials
-
animal enzyme: activated, bacterial enzyme: unaffected or inhibited
-
peroxynitrite
-
0.1 mM, 50% inhibition of cysteine-free mutant C85S/C152E, no inhibition of wild-type
[(5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(1-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-8-yl]ethyl)amino]acetic acid
-
[amino-(4-[[(amino-thioureido-methyl)-amino]-methyl]-2,5-dimethyl-benzylamino)-methyl]-thiourea
-
IC50: 109 nM, 0.03 mM 7,8-dihydrofolate
methotrexate
enzyme-bound crystal structure analysis for inhibitor screening and docking studies, PDB ID 1RA3
additional information
-
overview: design of inhibitors from X-ray crystal structures
-
additional information
-
inhibitory effect increased with the radius of the cations
-
additional information
inhibitory potencies of diverse 2,4-pteridinediamine derivates and pyrimido[4,5-d]pyrimidine-2,4-diamine derivates, molecular dynamic simulations, detailed overview
-
additional information
7H-pyrrolo(3,2-f) quinazoline-1, 3-diamine (PQD) inhibition kinetics, overview
-
additional information
analysis of 1,3,5-triazine-2,4-diamine and 1,2,4-triazine-2,4-diamine group-containing compounds as potential inhibitors of Escherichia coli DHFR. Fifteen derivatives of 1,3,5-triazine-2,4-diamine are assessed for their ability to bind to the apo form of enzyme EcDHFR. Quantitative structure-activity relationship analysis and macromolecular docking of the diaminotriazine series of compounds, overview. Effects of 1,3,5-triazine-2,4-diamine derivatives on the cofactor NADPH binding
-
additional information
-
analysis of 1,3,5-triazine-2,4-diamine and 1,2,4-triazine-2,4-diamine group-containing compounds as potential inhibitors of Escherichia coli DHFR. Fifteen derivatives of 1,3,5-triazine-2,4-diamine are assessed for their ability to bind to the apo form of enzyme EcDHFR. Quantitative structure-activity relationship analysis and macromolecular docking of the diaminotriazine series of compounds, overview. Effects of 1,3,5-triazine-2,4-diamine derivatives on the cofactor NADPH binding
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.003
10-formyl-dihydrofolate
pH 7.5, temperature not specified in the publication
0.0004
7,8-dihydrofolate
chimera with Haemophilus influenzae enzyme, pH 7, 25°C
0.0007
7,8-dihydrofolate
-
mutant enzyme R44H, at 15°C, pH not specified in the publication
0.0007
7,8-dihydrofolate
chimera with Listeria grayi enzyme, pH 7, 25°C
0.0008
7,8-dihydrofolate
deletion mutant DELTAAla145, at pH 7.0 and 25°C
0.0008
7,8-dihydrofolate
-
mutant enzyme R44V, at 15°C, pH not specified in the publication
0.001
7,8-dihydrofolate
-
mutant enzyme R44Y, at 15°C, pH not specified in the publication
0.0011
7,8-dihydrofolate
-
mutant enzyme R44C, at 15°C, pH not specified in the publication
0.0014
7,8-dihydrofolate
-
mutant enzyme R44L, at 15°C, pH not specified in the publication
0.0015
7,8-dihydrofolate
-
mutant enzyme R44G, at 15°C, pH not specified in the publication
0.0015
7,8-dihydrofolate
chimera with Saccharomyces cerevisiae enzyme, pH 7, 25°C
0.0016
7,8-dihydrofolate
-
mutant enzyme R44A, at 15°C, pH not specified in the publication
0.0016
7,8-dihydrofolate
-
wild type enzyme, at 15°C, pH not specified in the publication
0.0019
7,8-dihydrofolate
-
mutant enzyme R44E, at 15°C, pH not specified in the publication
0.0021
7,8-dihydrofolate
-
mutant enzyme R44I, at 15°C, pH not specified in the publication
0.0023
7,8-dihydrofolate
-
mutant enzyme R44M, at 15°C, pH not specified in the publication
0.0024
7,8-dihydrofolate
-
mutant enzyme R44Q, at 15°C, pH not specified in the publication
0.0025
7,8-dihydrofolate
-
mutant enzyme R44K, at 15°C, pH not specified in the publication
0.0025
7,8-dihydrofolate
-
mutant enzyme R44N, at 15°C, pH not specified in the publication
0.0025
7,8-dihydrofolate
-
mutant enzyme R44T, at 15°C, pH not specified in the publication
0.0027
7,8-dihydrofolate
-
mutant enzyme R44F, at 15°C, pH not specified in the publication
0.0027
7,8-dihydrofolate
-
pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface
0.0029
7,8-dihydrofolate
-
mutant enzyme R44P, at 15°C, pH not specified in the publication
0.003
7,8-dihydrofolate
mutant M1A/M16F/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0
0.003
7,8-dihydrofolate
-
mutant enzyme R44D, at 15°C, pH not specified in the publication
0.0031
7,8-dihydrofolate
-
mutant enzyme R44S, at 15°C, pH not specified in the publication
0.0037
7,8-dihydrofolate
-
mutant enzyme R44W, at 15°C, pH not specified in the publication
0.0038
7,8-dihydrofolate
mutant M1A/M16S/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0
0.0048
7,8-dihydrofolate
deletion mutant DELTAGly67/DELTAAla145, at pH 7.0 and 25°C
0.0051
7,8-dihydrofolate
deletion mutant DELTAGly67, at pH 7.0 and 25°C
0.0055
7,8-dihydrofolate
pH 7.5, temperature not specified in the publication
0.0056
7,8-dihydrofolate
deletion mutant DELTAGly67/DELTAGly121, at pH 7.0 and 25°C
0.0063
7,8-dihydrofolate
chimera with Streptococcus dysgalactiae enzyme, pH 7, 25°C
0.0073
7,8-dihydrofolate
deletion mutant DELTAGly121/DELTAAla145, at pH 7.0 and 25°C
0.0077
7,8-dihydrofolate
-
pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface
0.0083
7,8-dihydrofolate
mutant M1A/M16N/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0
0.0083
7,8-dihydrofolate
mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S, 15°C, pH 7.0
0.0096
7,8-dihydrofolate
-
mutant isoform that carries the K32M mutation in gene copies 1 and 3 and the Q67H mutation in gene copies 2 and 4, pH 7.0, 30°C
0.0108
7,8-dihydrofolate
deletion mutant DELTAGly121, at pH 7.0 and 25°C
0.014
7,8-dihydrofolate
-
mutant carrying the K32M mutation in gene copies 1 and 3 with the Q67H mutation in all gene copies, pH 7.0, 30°C
0.0502
7,8-dihydrofolate
deletion mutant DELTAArg52/DELTAGly67, at pH 7.0 and 25°C
0.0512
7,8-dihydrofolate
deletion mutant DELTAArg52/DELTAGly121, at pH 7.0 and 25°C
0.058
7,8-dihydrofolate
chimera with Plasmodium falciparum enzyme, pH 7, 25°C
0.0777
7,8-dihydrofolate
deletion mutant DELTAArg52/DELTAAla145, at pH 7.0 and 25°C
0.1161
7,8-dihydrofolate
deletion mutant DELTAArg52, at pH 7.0 and 25°C
0.0074
NADPH
pH 7.5, temperature not specified in the publication, cosubstrate: 10-formyl-dihydrofolate
0.008
NADPH
pH 7.5, temperature not specified in the publication, cosubstrate: 7,8-dihydrofolate
0.011
NADPH
-
mutant carrying the K32M mutation in gene copies 1 and 3 with the Q67H mutation in all gene copies, pH 7.0, 30°C
0.0195
NADPH
-
pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface
0.0199
NADPH
-
pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface
0.029
NADPH
-
mutant isoform that carries the K32M mutation in gene copies 1 and 3 and the Q67H mutation in gene copies 2 and 4, pH 7.0, 30°C
additional information
additional information
-
Km of enzymes with decreased binding of folate and antagonists and of drug-sensitive enzymes
-
additional information
additional information
-
selenomethionine-containing mutants
-
additional information
additional information
Michaelis-Menten kinetics
-
additional information
additional information
Michaelis-Menten kinetics
-
additional information
additional information
analysis of steady-state kinetics at pH 9.5
-
additional information
additional information
-
analysis of steady-state kinetics at pH 9.5
-
additional information
additional information
transient kinetics investigation of enzyme
-
additional information
additional information
enzyme activity decreases with increasing pressure. The Km values for dihydrofolate and NADPH are slightly higher at 200 MPa than at atmospheric pressure. The hydride transfer step is insensitive to pressure, while the dissociation constants of substrates and cofactors increase with pressure
-
additional information
additional information
-
enzyme activity decreases with increasing pressure. The Km values for dihydrofolate and NADPH are slightly higher at 200 MPa than at atmospheric pressure. The hydride transfer step is insensitive to pressure, while the dissociation constants of substrates and cofactors increase with pressure
-
additional information
additional information
molecular dynamics and spectroscopic analysis of the enzyme in transition state based on residues Tyr100 and Tyr111, overview
-
additional information
additional information
kinetic behavior of the wild-type enzyme ecDHFR and enzyme mutants D27S, Y100F, and D27S/Y100F across an extended pH range, including pH 9.0. Competitive hydrogen to deuterium and hydrogen to tritium kinetic isotope effects (KIEs) on the second-order rate constant (kcat/Km) are measured for wild-type and mutant ecDHFR variants to assess the intrinsic KIE on the catalyzed hydride transfer. Presteady-state, Michaelis-Menten, and single-turnover kinetics
-
additional information
additional information
-
kinetic behavior of the wild-type enzyme ecDHFR and enzyme mutants D27S, Y100F, and D27S/Y100F across an extended pH range, including pH 9.0. Competitive hydrogen to deuterium and hydrogen to tritium kinetic isotope effects (KIEs) on the second-order rate constant (kcat/Km) are measured for wild-type and mutant ecDHFR variants to assess the intrinsic KIE on the catalyzed hydride transfer. Presteady-state, Michaelis-Menten, and single-turnover kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4.6
10-formyl-dihydrofolate
pH 7.5, temperature not specified in the publication
0.05
7,8-dihydrofolate
-
pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface
0.22
7,8-dihydrofolate
deletion mutant DELTAGly121/DELTAAla145, at pH 7.0 and 25°C
0.3
7,8-dihydrofolate
-
mutant enzyme R44P, at 15°C, pH not specified in the publication
0.37
7,8-dihydrofolate
deletion mutant DELTAArg52/DELTAGly121, at pH 7.0 and 25°C
0.38
7,8-dihydrofolate
deletion mutant DELTAGly67/DELTAGly121, at pH 7.0 and 25°C
0.49
7,8-dihydrofolate
-
mutant carrying the K32M mutation in gene copies 1 and 3 with the Q67H mutation in all gene copies, pH 7.0, 30°C
0.69
7,8-dihydrofolate
-
mutant enzyme R44D, at 15°C, pH not specified in the publication
0.89
7,8-dihydrofolate
-
mutant enzyme R44E, at 15°C, pH not specified in the publication
0.89
7,8-dihydrofolate
-
mutant enzyme R44Y, at 15°C, pH not specified in the publication
0.98
7,8-dihydrofolate
-
mutant enzyme R44L, at 15°C, pH not specified in the publication
1.15
7,8-dihydrofolate
-
mutant enzyme R44I, at 15°C, pH not specified in the publication
1.31
7,8-dihydrofolate
-
mutant enzyme R44V, at 15°C, pH not specified in the publication
1.39
7,8-dihydrofolate
-
mutant enzyme R44F, at 15°C, pH not specified in the publication
1.39
7,8-dihydrofolate
-
pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface
1.59
7,8-dihydrofolate
-
mutant enzyme R44H, at 15°C, pH not specified in the publication
1.7
7,8-dihydrofolate
-
mutant isoform that carries the K32M mutation in gene copies 1 and 3 and the Q67H mutation in gene copies 2 and 4, pH 7.0, 30°C
1.7
7,8-dihydrofolate
deletion mutant DELTAGly121, at pH 7.0 and 25°C
1.78
7,8-dihydrofolate
-
mutant enzyme R44G, at 15°C, pH not specified in the publication
1.9
7,8-dihydrofolate
-
mutant enzyme R44Q, at 15°C, pH not specified in the publication
2
7,8-dihydrofolate
-
mutant enzyme R44T, at 15°C, pH not specified in the publication
2.41
7,8-dihydrofolate
-
mutant enzyme R44W, at 15°C, pH not specified in the publication
2.43
7,8-dihydrofolate
-
mutant enzyme R44C, at 15°C, pH not specified in the publication
3.05
7,8-dihydrofolate
-
mutant enzyme R44A, at 15°C, pH not specified in the publication
3.3
7,8-dihydrofolate
-
mutant enzyme R44N, at 15°C, pH not specified in the publication
3.8
7,8-dihydrofolate
-
mutant enzyme R44M, at 15°C, pH not specified in the publication
5.4
7,8-dihydrofolate
-
wild type enzyme, at 15°C, pH not specified in the publication
5.7
7,8-dihydrofolate
-
mutant enzyme R44K, at 15°C, pH not specified in the publication
5.8
7,8-dihydrofolate
-
mutant enzyme R44S, at 15°C, pH not specified in the publication
10
7,8-dihydrofolate
chimera with Haemophilus influenzae enzyme, pH 7, 25°C
16.2
7,8-dihydrofolate
mutant M1A/M16S/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0
16.5
7,8-dihydrofolate
deletion mutant DELTAGly67, at pH 7.0 and 25°C
22.8
7,8-dihydrofolate
pH 7.5, temperature not specified in the publication
23.1
7,8-dihydrofolate
deletion mutant DELTAArg52/DELTAGly67, at pH 7.0 and 25°C
25.8
7,8-dihydrofolate
mutant M1A/M16F/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0
29
7,8-dihydrofolate
chimera with Saccharomyces cerevisiae enzyme, pH 7, 25°C
31.9
7,8-dihydrofolate
deletion mutant DELTAArg52/DELTAAla145, at pH 7.0 and 25°C
33
7,8-dihydrofolate
chimera with Plasmodium falciparum enzyme, pH 7, 25°C
37
7,8-dihydrofolate
chimera with Streptococcus dysgalactiae enzyme, pH 7, 25°C
40.5
7,8-dihydrofolate
mutant M1A/M16N/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0
40.5
7,8-dihydrofolate
mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S, 15°C, pH 7.0
41.1
7,8-dihydrofolate
deletion mutant DELTAGly67/DELTAAla145, at pH 7.0 and 25°C
96.3
7,8-dihydrofolate
deletion mutant DELTAArg52, at pH 7.0 and 25°C
0.05
NADPH
-
pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface
1.39
NADPH
-
pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface
4.05
NADPH
pH 7.5, temperature not specified in the publication, cosubstrate: 10-formyl-dihydrofolate
28.1
NADPH
pH 7.5, temperature not specified in the publication, cosubstrate: 7,8-dihydrofolate
additional information
additional information
-
selenomethionine-containing mutants
-
additional information
additional information
analysis of kinetic and thermodynamic fitting parameters and relaxation dispersion curves at different pH values and temperatures
-
additional information
additional information
-
analysis of kinetic and thermodynamic fitting parameters and relaxation dispersion curves at different pH values and temperatures
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1533.3
10-formyl-dihydrofolate
pH 7.5, temperature not specified in the publication
10
7,8-dihydrofolate
deletion mutant DELTAArg52/DELTAGly121, at pH 7.0 and 25°C
30
7,8-dihydrofolate
deletion mutant DELTAGly121/DELTAAla145, at pH 7.0 and 25°C
70
7,8-dihydrofolate
deletion mutant DELTAGly67/DELTAGly121, at pH 7.0 and 25°C
100
7,8-dihydrofolate
-
mutant enzyme R44P, at 15°C, pH not specified in the publication
160
7,8-dihydrofolate
deletion mutant DELTAGly121, at pH 7.0 and 25°C
230
7,8-dihydrofolate
-
mutant enzyme R44D, at 15°C, pH not specified in the publication
410
7,8-dihydrofolate
deletion mutant DELTAArg52/DELTAAla145, at pH 7.0 and 25°C
460
7,8-dihydrofolate
deletion mutant DELTAArg52/DELTAGly67, at pH 7.0 and 25°C
470
7,8-dihydrofolate
-
mutant enzyme R44E, at 15°C, pH not specified in the publication
500
7,8-dihydrofolate
-
mutant enzyme R44F, at 15°C, pH not specified in the publication
500
7,8-dihydrofolate
chimera with Plasmodium falciparum enzyme, pH 7, 25°C
560
7,8-dihydrofolate
-
mutant enzyme R44I, at 15°C, pH not specified in the publication
660
7,8-dihydrofolate
-
mutant enzyme R44W, at 15°C, pH not specified in the publication
680
7,8-dihydrofolate
-
mutant enzyme R44L, at 15°C, pH not specified in the publication
800
7,8-dihydrofolate
-
mutant enzyme R44Q, at 15°C, pH not specified in the publication
800
7,8-dihydrofolate
-
mutant enzyme R44T, at 15°C, pH not specified in the publication
900
7,8-dihydrofolate
-
mutant enzyme R44Y, at 15°C, pH not specified in the publication
1200
7,8-dihydrofolate
-
mutant enzyme R44G, at 15°C, pH not specified in the publication
1300
7,8-dihydrofolate
-
mutant enzyme R44N, at 15°C, pH not specified in the publication
1600
7,8-dihydrofolate
-
mutant enzyme R44M, at 15°C, pH not specified in the publication
1700
7,8-dihydrofolate
-
mutant enzyme R44V, at 15°C, pH not specified in the publication
1900
7,8-dihydrofolate
-
mutant enzyme R44A, at 15°C, pH not specified in the publication
1900
7,8-dihydrofolate
-
mutant enzyme R44S, at 15°C, pH not specified in the publication
2200
7,8-dihydrofolate
-
mutant enzyme R44C, at 15°C, pH not specified in the publication
2300
7,8-dihydrofolate
-
mutant enzyme R44K, at 15°C, pH not specified in the publication
2500
7,8-dihydrofolate
-
mutant enzyme R44H, at 15°C, pH not specified in the publication
3300
7,8-dihydrofolate
deletion mutant DELTAGly67, at pH 7.0 and 25°C
3400
7,8-dihydrofolate
-
wild type enzyme, at 15°C, pH not specified in the publication
4560
7,8-dihydrofolate
pH 7.5, temperature not specified in the publication
6000
7,8-dihydrofolate
chimera with Streptococcus dysgalactiae enzyme, pH 7, 25°C
8600
7,8-dihydrofolate
deletion mutant DELTAGly67/DELTAAla145, at pH 7.0 and 25°C
22000
7,8-dihydrofolate
chimera with Listeria grayi enzyme, pH 7, 25°C
24000
7,8-dihydrofolate
chimera with Saccharomyces cerevisiae enzyme, pH 7, 25°C
25000
7,8-dihydrofolate
chimera with Haemophilus influenzae enzyme, pH 7, 25°C
29400
7,8-dihydrofolate
deletion mutant DELTAAla145, at pH 7.0 and 25°C
547.3
NADPH
pH 7.5, temperature not specified in the publication, cosubstrate: 10-formyl-dihydrofolate
3512.5
NADPH
pH 7.5, temperature not specified in the publication, cosubstrate: 7,8-dihydrofolate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0000115
1,4-phenylenebis(methylene) bis(N-[amino(imino)methyl](imidothiocarbamate))
25°C, pH 7.5
0.01116
1-(3-ethoxyphenyl)-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
pH 7.3, 22°C
0.00102
1-(4-chlorophenyl)-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
pH 7.3, 22°C
0.00402
1-(4-ethoxyphenyl)-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
pH 7.3, 22°C
0.02022
1-[3-(aminomethyl)phenyl]-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
pH 7.3, 22°C
0.0008802
1-[3-chloro-4-(5-phenylpentyl)phenyl]-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
pH 7.3, 22°C
0.0393
1-[4-(aminomethyl)phenyl]-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
pH 7.3, 22°C
0.000958
3-(4,6-diamino-2,2-dimethyl-1,3,5-triazin-1(2H)-yl)benzonitrile
pH 7.3, 22°C
0.00289
4-[3-(4,6-diamino-2,2-dimethyl-1,3,5-triazin-1(2H)-yl)benzamido]benzene-1-sulfonyl fluoride
pH 7.3, 22°C
0.105
4-[4,6-diamino-2-(4-chlorophenyl)-1,3,5-triazin-1(2H)-yl]benzene-1-sulfonamide
pH 7.3, 22°C
0.0000497
4-[4-[3-(4,6-diamino-2,2-dimethyl-1,3,5-triazin-1-yl)phenyl]butyl]benzenesulfonyl fluoride
pH 7.3, 22°C
0.0000926
4-[6-[4-(4,6-diamino-2,2-dimethyl-1,3,5-triazin-1-yl)phenyl]hexyl]benzenesulfonyl fluoride
pH 7.3, 22°C
0.00155
6,6-dimethyl-1-(4-methylphenyl)-1,6-dihydro-1,3,5-triazine-2,4-diamine
pH 7.3, 22°C
0.0003591
6,6-dimethyl-1-[3-(4-phenylbutyl)phenyl]-1,6-dihydro-1,3,5-triazine-2,4-diamine
pH 7.3, 22°C
0.00000742
7-[(4-aminophenyl)methyl]-7H-pyrrolo[3,2-f]quinazoline-1,3-diamine
pH 7.3, 22°C
additional information
additional information
-
Ki of substrates 7,8-dihydrofolate and NADPH for the 2 isozymes
-
additional information
additional information
-
Ki of substrates dihydrofolate, NADPH and acetylpyridine adenine nucleotide
-
additional information
additional information
inhibition kinetics, overview
-
additional information
additional information
-
inhibition kinetics, overview
-
additional information
additional information
inhibition kinetics, overview
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0068
(4-(1-[(5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(methyl)amino]ethyl)phenyl)methanol
Escherichia coli
pH 7.0, 37°C
0.000000011
1,6-bis-(4-fluoro-phenyl)-[1,3,5]triazine-2,4-diamine
Escherichia coli
-
IC50: 0.011 nM, 0.03 mM 7,8-dihydrofolate
0.019
1-(3-([(2,4-diaminopteridin-6-yl)methyl]amino)phenyl)ethanol
Escherichia coli
pH 7.0, 37°C
0.263
1-(3-ethoxyphenyl)-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
Escherichia coli
pH 7.3, 22°C
0.02091
1-(4-chlorophenyl)-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
Escherichia coli
pH 7.3, 22°C
0.0891
1-(4-ethoxyphenyl)-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
Escherichia coli
pH 7.3, 22°C
0.478
1-[3-(aminomethyl)phenyl]-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
Escherichia coli
pH 7.3, 22°C
0.01845
1-[3-chloro-4-(5-phenylpentyl)phenyl]-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
Escherichia coli
pH 7.3, 22°C
0.722
1-[4-(aminomethyl)phenyl]-6,6-dimethyl-1,6-dihydro-1,3,5-triazine-2,4-diamine
Escherichia coli
pH 7.3, 22°C
0.0017
2-(2-((5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(1-(naphthalen-1-yl)ethyl)amino)ethoxy)ethanol
Escherichia coli
pH 7.0, 37°C
0.000028
2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.028
2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.000033
2-amino-5-[(2-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.0028
2-amino-5-[(3,4-dichlorophenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.000022
2-amino-5-[(3,4-dichlorophenyl)thio]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.0000028
2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]-pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.0028
2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.000027
2-amino-5-[(3,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.000003
2-amino-5-[(3-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.000029
2-amino-5-[(4-bromophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.000042
2-amino-5-[(4-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.028
2-amino-5-[(4-chlorophenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.032
2-amino-5-[(4-fluorophenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.000028
2-amino-6-ethyl-5-(2-naphthylthio)thieno[2,3-d ]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.000033
2-amino-6-ethyl-5-(phenylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.000033
2-amino-6-ethyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d ]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.000031
2-amino-6-ethyl-5-[(4-fluorophenyl)sulfanyl]thieno[2,3-d ]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.000039
2-amino-6-ethyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
pH 7.0, 37°C
0.058
2-amino-6-methyl-5-(2-naphthylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.035
2-amino-6-methyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0012
2-amino-6-methyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0003
2-[(5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(1-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-8-yl]ethyl)amino]ethanol
Escherichia coli
pH 7.0, 37°C
0.076
2-[(5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(methyl)amino]-2-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-yl]ethanol
Escherichia coli
pH 7.0, 37°C
0.00093
3-((5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(1-(naphthalen-1-yl)ethyl)amino)propanoic acid
Escherichia coli
pH 7.0, 37°C
0.02019
3-(4,6-diamino-2,2-dimethyl-1,3,5-triazin-1(2H)-yl)benzonitrile
Escherichia coli
pH 7.3, 22°C
0.48
3-heptyl-4-hydroxy-1H-naphthalen-2-one
Escherichia coli
-
IC50: 0.48 mM, 0.03 mM 7,8-dihydrofolate
0.00273
4-((5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(1-(naphthalen-1-yl)ethyl)amino)butanoic acid
Escherichia coli
pH 7.0, 37°C
0.0002
4-([(2,4-diaminopteridin-6-yl)methyl](methyl)amino)benzoic acid
Escherichia coli
pH 7.0, 37°C
0.06283
4-[3-(4,6-diamino-2,2-dimethyl-1,3,5-triazin-1(2H)-yl)benzamido]benzene-1-sulfonyl fluoride
Escherichia coli
pH 7.3, 22°C
1.548
4-[4,6-diamino-2-(4-chlorophenyl)-1,3,5-triazin-1(2H)-yl]benzene-1-sulfonamide
Escherichia coli
pH 7.3, 22°C
0.00115
4-[4-[3-(4,6-diamino-2,2-dimethyl-1,3,5-triazin-1-yl)phenyl]butyl]benzenesulfonyl fluoride
Escherichia coli
pH 7.3, 22°C
0.00184
4-[6-[4-(4,6-diamino-2,2-dimethyl-1,3,5-triazin-1-yl)phenyl]hexyl]benzenesulfonyl fluoride
Escherichia coli
pH 7.3, 22°C
0.000018
5-(3,4,5-trimethoxy-benzyl)-pyrimidine-2,4-diamine
Escherichia coli
-
IC50: 18 nM, 0.03 mM 7,8-dihydrofolate
0.03227
6,6-dimethyl-1-(4-methylphenyl)-1,6-dihydro-1,3,5-triazine-2,4-diamine
Escherichia coli
pH 7.3, 22°C
0.00759
6,6-dimethyl-1-[3-(4-phenylbutyl)phenyl]-1,6-dihydro-1,3,5-triazine-2,4-diamine
Escherichia coli
pH 7.3, 22°C
0.00079
6,7-dimethyl-5,6,7,8-tetrahydro-quinazoline-2,4-diamine
Escherichia coli
-
IC50: 790 nM, 0.03 mM 7,8-dihydrofolate
0.0006
6-(([4-chlorotricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-yl]amino)methyl)pteridine-2,4-diamine
Escherichia coli
pH 7.0, 37°C
0.00066
6-(4-trifluoromethyl-phenoxy)-quinazoline-2,4-diamine
Escherichia coli
-
IC50: 660 nM, 0.03 mM 7,8-dihydrofolate
0.1
6-([(4-aminophenyl)amino]methyl)pteridine-2,4-diamine
Escherichia coli
pH 7.0, 37°C
0.014
6-([(4-ethoxyphenyl)amino]methyl)pteridine-2,4-diamine
Escherichia coli
pH 7.0, 37°C
0.0006
6-([butyl(phenyl)amino]methyl)pteridine-2,4-diamine
Escherichia coli
pH 7.0, 37°C
0.00011
6-([ethyl(phenyl)amino]methyl)pteridine-2,4-diamine
Escherichia coli
pH 7.0, 37°C
0.00015
6-([phenyl(prop-2-en-1-yl)amino]methyl)pteridine-2,4-diamine
Escherichia coli
pH 7.0, 37°C
0.00088
6-([phenyl(propan-2-yl)amino]methyl)pteridine-2,4-diamine
Escherichia coli
pH 7.0, 37°C
0.00015
6-([phenyl(propyl)amino]methyl)pteridine-2,4-diamine
Escherichia coli
pH 7.0, 37°C
0.011
6-([tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-ylamino]methyl)pteridine-2,4-diamine
Escherichia coli
pH 7.0, 37°C
0.048
6-([tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-8-ylamino]methyl)pteridine-2,4-diamine
Escherichia coli
pH 7.0, 37°C
0.00019
6-p-tolyloxy-quinazoline-2,4-diamine
Escherichia coli
-
IC50: 190 nM, 0.03 mM 7,8-dihydrofolate
0.00031
6-p-tolylsulfanyl-quinazoline-2,4-diamine
Escherichia coli
-
IC50: 310 nM, 0.03 mM 7,8-dihydrofolate
0.1
6-[(tricyclo[4.3.0.07,9]nona-1,3,5-trien-3-ylamino)methyl]pteridine-2,4-diamine
Escherichia coli
pH 7.0, 37°C
0.000000011
7-methyl-5,6,7,8-tetrahydro-quinazoline-2,4-diamine
Escherichia coli
-
IC50: 0.011 nM, 0.03 mM 7,8-dihydrofolate
0.0022
methyl 4-([(2,4-diaminopteridin-6-yl)methyl](methyl)amino)benzoate
Escherichia coli
pH 7.0, 37°C
0.00032
N-(4-chloro-2-cyanophenyl)imidodicarbonimidic diamide
Escherichia coli
-
IC50: 320 nM, 0.03 mM 7,8-dihydrofolate
0.000001
N-(4-[(2-amino-6-ethyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid
Escherichia coli
-
pH 7.0, 37°C
0.0000002
N-(4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid
Escherichia coli
-
pH 7.0, 37°C
0.0002
N-(4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d]pyrimidin-5-yl)sulfanyl]benzoyl)-L-glutamic acid
Escherichia coli
-
-
0.0083
N-(5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)-N-[1-(2-ethoxynaphthalen-1-yl)ethyl]-b-alanine
Escherichia coli
pH 7.0, 37°C
0.00096
N7-(1-(naphthalen-1-yl)ethyl)-N7-propylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.002
N7-(2-aminoethyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.001
N7-(2-ethoxyethyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0033
N7-(2-propoxyethyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0035
N7-(3-ethoxypropyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0048
N7-(4-aminobutyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.003
N7-(5-aminopentyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0006
N7-(cyclopropylmethyl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0035
N7-(cyclopropylmethyl)-N7-[1-(4-methylnaphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0004
N7-(prop-2-en-1-yl)-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.00088
N7-benzyl-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0357
N7-benzyl-N7-methylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.00184
N7-butyl-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.004
N7-cyclopropyl-N7-(1-(naphthalen-2-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0002
N7-ethyl-N7-(1-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-yl]ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0032
N7-ethyl-N7-(1-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-8-yl]ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0214
N7-ethyl-N7-(4-methylbenzyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.00021
N7-ethyl-N7-[1-(4-methylnaphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0112
N7-methyl-N7-(1-phenylethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.000016
N7-methyl-N7-(1-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-yl]ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.000055
N7-methyl-N7-[1-(4-methylnaphthalen-1-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0007
N7-methyl-N7-[1-(quinolin-4-yl)ethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0035
N7-methyl-N7-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-7-ylmethyl]pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.00185
N7-pentyl-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.00027
N7-[1-(1-benzothiophen-3-yl)ethyl]-N7-(cyclopropylmethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.00015
N7-[1-(1-benzothiophen-3-yl)ethyl]-N7-ethylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.00005
N7-[1-(1-benzothiophen-3-yl)ethyl]-N7-methylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0011
N7-[1-(2-ethoxynaphthalen-1-yl)ethyl]-N7-methylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.003
N7-[1-(2-ethylnaphthalen-1-yl)ethyl]-N7-methylpyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0022
N7-[1-(4-chloronaphthalen-1-yl)ethyl]-N7-(cyclopropylmethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.00103
N7-[2-(4-fluorophenyl)ethyl]-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0025
N7-[2-(cyclohex-2-en-1-yl)ethyl]-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0008
N7-[3-(2-methyl-4H-imidazol-4-yl)propyl]-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.0034
N7-[3-(2-methylpropoxy)propyl]-N7-(1-(naphthalen-1-yl)ethyl)pyrimido[4,5-d]pyrimidine-2,4,7-triamine
Escherichia coli
pH 7.0, 37°C
0.021
[(5,7-diaminopyrimido[4,5-d]pyrimidin-2-yl)(1-[tricyclo[4.4.0.02,5]deca-1(10),2(5),6,8-tetraen-8-yl]ethyl)amino]acetic acid
Escherichia coli
pH 7.0, 37°C
0.000109
[amino-(4-[[(amino-thioureido-methyl)-amino]-methyl]-2,5-dimethyl-benzylamino)-methyl]-thiourea
Escherichia coli
-
IC50: 109 nM, 0.03 mM 7,8-dihydrofolate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
45
additional information
additional information
-
ecDHFR shows a monotonous decrease of its activity with increasing pressure, down to 50% at 250 MPa
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.5
6.5
7.6
additional information
additional information
-
trimethoprim-resistant, form 2: no detectable pH-optimum, continuous increase of activity as pH decreases from pH 9.0 to pH 4.0
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.2 - 8.8
-
half-maximal activities at pH 4.2 and pH 8.8, acetate buffer, Tris buffer
additional information
additional information
analysis of the pH dependence of wild-type and mutant enzymes, overview. The differences between the temperature-independent deuteride transfer rate constant Dkhyd at pH 9.0 and temperature-dependent Dkhyd at pH 7.0 actually result from a pH-dependent commitment to catalysis on the khyd
additional information
-
analysis of the pH dependence of wild-type and mutant enzymes, overview. The differences between the temperature-independent deuteride transfer rate constant Dkhyd at pH 9.0 and temperature-dependent Dkhyd at pH 7.0 actually result from a pH-dependent commitment to catalysis on the khyd
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
37
additional information
additional information
strong dependence of reaction rates on temperature. Temperature-independent kinetic isotope effects between 15 and 35°C
additional information
-
strong dependence of reaction rates on temperature. Temperature-independent kinetic isotope effects between 15 and 35°C
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
-
replacement of Asp27 in Escherichia coli DHFR by Glu27 in Moritella DHFR may be an adaptation for cold that fortuitously also enhances activity under pressure. The extra carbon of a glutamate increases flexibility of the Thr113-Res27 hydrogen bond while pressure increases the hydrogen bond strength and correlation of sheet F with helix B
malfunction
growth deficiency phenotypes (in un-supplemented M9 minimal medium containing thymidine) are direct consequences of the gene deletions
metabolism
physiological function
for mechanism, dynamics are crucial for solvent entry and protonation of substrate. The mechanism invokes the release of a sole proton from a hydronium (H3O+) ion, its pathway through a narrow channel that sterically hinders the passage of water, and the ultimate protonation of DHF at the N5 atom. DOD47 is the catalytic water that promotes protonation of the N5 atom in DHF. The deuteron, modeled into the nuclear difference density peak in the active site, possibly forms a low-barrier hydrogen bond with the oxygen atom of DOD47 and is positioned between the Met20 side chain and the DOD47 so as to define a pathway that could lead to protonation of N5 by the deuteron
additional information
metabolism
enzyme DHFR is a pivotal enzyme in the thymidine and purine synthesis pathway
metabolism
study of protein dynamics, using a pump-probe method that employs pulsed-laser photothermal heating of a gold nanoparticle (AuNP) to directly excite a local region of the protein structure and transient absorbance to probe the effect on enzyme activity. Activity is accelerated by pulsed-laser excitation when the AuNP is attached close to a network of coupled motions in DHFR. No rate acceleration is observed when the AuNP is attached away from the network with pulsed excitation, or for any attachment site with continuous wave excitation
additional information
analysis of the orientation of residues Y100, D27, M20, and the ligands in the active site of ecDHFR, PDB ID 1rx2
additional information
-
analysis of the orientation of residues Y100, D27, M20, and the ligands in the active site of ecDHFR, PDB ID 1rx2
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). PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
17300 - 18300
18000
17300 - 18300
-
trimethoprim-resistant, amino acid composition, sedimentation velocity
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
monomer
monomer
-
preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
for equilibrium and kinetic folding mechanism, a continuous adenosine-binding domain is required for a stable thermal intermediate and a continuous discontinuous loop domain is required for a stable urea intermediate
additional information
-
for equilibrium and kinetic folding mechanism, a continuous adenosine-binding domain is required for a stable thermal intermediate and a continuous discontinuous loop domain is required for a stable urea intermediate
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
structure of plasmid-encoded R67 isoform bound to NADP+, at 1.15 A resolution. NADP+ assumes an extended conformation with the nicotinamide ring bound near the center of the active site pore, the ribose and diphospate group extending toward the outer pore. the ribonicotinamide exhibits anti conformation, and the coenzyme displayes symmetrical binding mode with several water-mediated hydrogen bonds
ternary complex of isoform R67 with dihydrofolate and NADPH, resolved to 1.26 A. In the catalytic complex, the polar backbone atoms of two symmetry-related I68 residues provide recognition motifs that interact with the carboxamide on the nicotinamide ring, and the N3-O4 amide function on the pteridine ring. This set of interactions orients the aromatic rings of substrate and cofactor in a relative endo geometry in which the reactive centers are held in close proximity. Additionally, a central, hydrogen-bonded network consisting of two pairs of Y69-Q67/Q67-Y69 residues provides a tight interface, which holds the substrates in place in an orientation conductive to hydride transfer
2.1 A resolution neutron structure of a pseudo-Michaelis complex determined at acidic pH, direct observation of the catalytic proton and its parent solvent molecule
analysis of higher energy conformational substrates by NMR relaxation dispersion. The maximum hydride transfer and steady-state turnover rates are governed by the dynamics of transitions between ground and excited states of the intermediates. Model of conformational changes during the catalytic cycle
comparison of temperature dependence of dynamics between Geobacillus stearothermophilus and Escherichia coli enzymes using elastic coherent scattering. The Geobacillus' enzyme has a significantly broader distribution and slightly larger amplitudes of the atomic mean-square displacements extracted from the dynamic structure factor
crystal structure of seleno- and telluromethionine-containing enzyme in complex with methotrexate and CaCl2 by vapour diffusion hanging drop method, X-ray analysis with synchotron and rotating anode generator as X-ray source
in complex with inhibitor 1,4-phenylenebis(methylene) bis(N-[amino(imino)methyl](imidothiocarbamate)) and NADPH. Inhibitor exploits a unique binding surface in loop M20
in the excited state of the enzyme:THF:NADPH product release complex, the reduced nicotinamide ring of the cofactor transiently enters the active site where it displaces the pterin ring of the THF product. The p-aminobenzoyl-L-glutamate tail of THF remains weakly bound in a widened binding cleft
microsecond compaction dynamics study of the folding. A significant collapse of the radius of gyration from 30 A to 23.2 A occurs within 0.3 sec after the initiatiation of refolding by a urea dilution jump. the subsequent folding occurs on a considerably longer time scale. Experimental data may best be explained by the specific hydrophobic collapse model. The folding trajectory of the protein is located between those of alpha-helical and beta-sheet proteins
modeling of complexes with inhibitors hydrochlorothiazide and indapamide. Compared to the DHFR-ethacrynic acid complex, DHFR-sulphadiazine, DHFR-indapamide, and DHFR-hydrochlorothiazide complexes have higher negative binding energies. The compounds may act similar to antibiotics from the perspective of binding with DHFR
mutant M1A/M16N/M20L/M42Y/M92F/C85A/C152S, increased structural flexibility and an increased size of the N-(p-aminobenzoyl)-L-glutamate binding cleft
self-organized polymer model to monitor the kinetics of closed to occluded and reverse transitions. During the closed to occluded transition, coordinated changes in a number of residues in the loop domain enable the M20 loop to slide along the alpha-helix in the adenosine-binding domain. Sliding is triggered by pulling of the M20 loop by the betaG-betaH loop and the pushing action of the betaG-betaH loop. The residues that facilitate the M20 loop motion are part of the network of residues that transmit allosteric signals during the close to occluded transition. Replacement of M16 and G121 by a disulfide cross-link impedes that transition. The order of events in the occluded to closed transition is not the reverse of the forward transition. The contact E18-S49 in the occluded structure persists until the sliding of the M20 loop is nearly complete
structures of protein crystallized in varying ionic strengths. High ionic strengths (0.75/1.5M) can preferentially stabilize the loop in closed/occluded conformations
study on enzyme ternary complex with substrate analogue folates and oxidized NADP+ cofactor using NMR relaxation methods. Conformational exchanges of protein between a ground state with closed conformation of active site loops and an excited state with loops in occluded conformation. Fluctuations include motions of the nicotinamide ring of the cofactor into and out of the active site
use of carbon-deuterium bonds as probes of proteins. The stretching absorption frequency of (methyl-d3) methionine carbon-deuterium bonds shows an approximately linear dependence on solvent dielectric. Characterization of the IR absorptions at residues Met16 and Met20, within the catalytically important Met20 loop, and Met42, which is located within the hydrophobic core of the enzyme. The carbon-deuterium bonds tare sensitive to their local protein environment and dihydrofolate reductase is electrostatically and dynamically heterogeneous
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
I68M
compared to wild-type, low kcat/Km (DHF) values. Mutant allows growth in presence of sorbitol up to 1.44 osmol conditions
Q67H
mutant with reasonable catalytic efficiency, but displays substrate and cofactor inhibition. Contrary to wild-type which allows growth on all sorbitol conditions until the osmolyte concentration becomes too high or cell water content becomes too low, mutant Q67H allows growth up to 1.81 osmol conditions
Y69L
compared to wild-type, low kcat/Km (DHF) values. Mutant allows growth in presence of sorbitol up to 0.81 osmol conditions
C85S/C152E
-
cysteine-free mutant, analysis of inhibition by p.eroxynitrite. bicarbonate buffer protects mutant from inhibition by peroxynitrite. Decrease in mutant stability upon oxidation
D27S
site-directed mutagenesis, the mutant shows a 3400fold reduced rate for the NADPH-dependent reduction of 7,8-dihydrofolate at pH 7.0 in water compared to wild-type
DELTAAla145
the mutant shows increased catalytic efficiency compared to the wild type enzyme
DELTAArg52
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
DELTAArg52/DELTAAla145
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
DELTAArg52/DELTAGly121
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
DELTAArg52/DELTAGly67
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
DELTAGly121
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
DELTAGly121/DELTAAla145
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
DELTAGly67
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
DELTAGly67/DELTAAla145
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
DELTAGly67/DELTAGly121
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
G121V
I14A
the mutant shows strongly reduced pre-steady-state rates of H transfer at 25°C and pH 7.0 compared to the wild type enzyme
I14G
the mutant shows severely reduced pre-steady-state rates of H transfer at 25°C and pH 7.0 compared to the wild type enzyme
I14V
the mutant shows reduced pre-steady-state rates of H transfer at 25°C and pH 7.0 compared to the wild type enzyme
K32M
-
weakens binding of dihydrofolate over 60-fold, increases kcat value by a factor of 5
L16M
-
site-directed mutagenesis, same kinetic properties like the wild-type enzyme
L16M/L20M
-
site-directed mutagenesis, double mutant, elevated turnover number and specific activity
L16M/L20M/L42M
-
site-directed mutagenesis, triple mutant, elevated turnover number and specific activity
L16SeM
-
site-directed mutagenesis, with selenomethionine, same kinetic properties like the wild-type enzyme
L16SeM/L20SeM
-
site-directed mutagenesis, double mutant, with selenomethionine, elevated turnover number and specific activity
L16SeM/L20SeM/L42SeM
-
site-directed mutagenesis, triple mutant, with selenomethionine, elevated turnover number and specific activity
L20M
-
site-directed mutagenesis, elevated turnover number and specific activity
L20SeM
-
site-directed mutagenesis, with selenomethionine, elevated turnover number and specific activity
L42M
-
site-directed mutagenesis, same kinetic properties like the wild-type enzyme
L42SeM
-
site-directed mutagenesis, with selenomethionine, same kinetic properties like the wild-type enzyme
L54I
-
the mutation reduces the hydride transfer efficiency by about 100fold
L92M
-
site-directed mutagenesis, same kinetic properties like the wild-type enzyme
L92SeM
-
site-directed mutagenesis, with selenomethionine, same kinetic properties like the wild-type enzyme
M1A/M16F/M20L/M42Y/M92F/C85A/C152S
hyperactive mutant, increase in dissociation rate constant of tetrahydrofolate from the enzyme-NADPH-tetrahydrofolate ternary complex
M1A/M16N/M20L/M42Y/C85A/M92F/C152S
mutant ANLYF carries seven amino acid substitutions that result in a methionine- and cysteine-free mutant enzyme with decreased stability. Mutant ANLYF show very high activity and may be stabilized by backbone cyclization via a cyanocysteine-mediated intramolecular ligation reaction, without loss of its high activity
M1A/M16N/M20L/M42Y/M92F/C85A/C152S
hyperactive mutant, increase in dissociation rate constant of tetrahydrofolate from the enzyme-NADPH-tetrahydrofolate ternary complex
M1A/M16S/M20L/M42Y/M92F/C85A/C152S
hyperactive mutant, increase in dissociation rate constant of tetrahydrofolate from the enzyme-NADPH-tetrahydrofolate ternary complex
M42A
-
45% decrease in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42C
-
slight decrease in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42E
-
6fold decrease in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42G
-
45% decrease in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42H
-
40% decrease in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42I
-
slight increase in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42L
-
slight decrease in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42P
-
50% increase in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42Q
-
45% decrease in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42S
-
45% decrease in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42T
-
slight increase in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42V
-
slight decrease in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42W
M42W/G121V
-
kinetic isotope effect study, major change in the nature of H transfer, leading to poor reorganization and substantial gating
M42Y
-
30% increase in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
Q67H
-
about 100-fold tightening in binding to both dihydrolfolate and NADPH, mutation can help rescue the K32 M mutation
R44A
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44C
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44D
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44E
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44F
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44G
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44I
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44K
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44L
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44M
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44N
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44P
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44Q
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44S
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44T
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44V
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44W
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
R44Y
-
the mutation significantly reduces the enzymatic activity and the binding affinity toward the cofactor NADPH
Y100F
site-directed mutagenesis, the mutant shows a 14fold reduced rate for the NADPH-dependent reduction of 7,8-dihydrofolate at pH 7.0 in water compared to wild-type
Y100F/D27S
site-directed mutagenesis, the mutant shows an over 100000fold reduced rate for the NADPH-dependent reduction of 7,8-dihydrofolate at pH 7.0 in water compared to wild-type
additional information
G121V
the mutation interferes with coupled loop movements and appears to impair catalysis by destabilizing the closed Michaelis complex and introducing an extra step into the kinetic pathway
G121V
-
kinetic isotope effect study, environmentally coupled tunnelling similar to wild-type
G121V
reduced catalytic efficiency due to significantly reduced pre-exponential factors, the chemical mechanism of catalysis is not impaired
G121V
-
the mutation reduces the hydride transfer efficiency by about 100fold
M42W
-
6fold decrease in ratio kcat/Km, analysis of thermodynamic parameters for urea denaturation
M42W
-
kinetic isotope effect study, environmentally coupled tunnelling similar to wild-type
additional information
double-labeling of enzyme with fluorescent quencher QSY35, at amino acid 17, and fluorescent probe Alexa555,at amino acid 35, by introducing Cys residues at these sites. Labeled enzyme retains full catalytic activity, analysis of first order rate constants for the stopped-flow fluorescent change of forward and reverse reaction
additional information
permutations in the N- and C-terminal domain show that for equilibrium and kinetic folding mechanism, a continuous adenosine-binding domain is required for a stable thermal intermediate and a continuous discontinuous loop domain is required for a stable urea intermediate
additional information
-
permutations in the N- and C-terminal domain show that for equilibrium and kinetic folding mechanism, a continuous adenosine-binding domain is required for a stable thermal intermediate and a continuous discontinuous loop domain is required for a stable urea intermediate
additional information
-
construction of a mutant isoform that carries the K32M mutation in gene copies 1 and 3 and the Q67H mutation in gene copies 2 and 4 of the tetrameric enzyme. Mutation Q67H can mostly compensate for mutation K32M. In a multimutant that combines the K32M mutation in gene copies 1 and 3 with the Q67H mutation in all gene copies, dihydrolflate is inhibitory, while NADPH is not. A mutant containing only the Q67H substitution in all four gene copies is severely inhibited by both NADPH and substrate
additional information
construction of mutants based on mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S, i.e. ANLYF. Linear ANLYF-Gn is ANLYF with the N- and C-terminal extensions of Ala-(Gly)n-3 and -(Gly)2-Cys-Ala residues, respectively, with n=6-8, and circular ANLYF-Gn is ANLYF with the C and N termini cross-linked by the linker of the -Gly-Gly-Ala-(Gly)n-3 sequence, n=6-8
additional information
-
construction of mutants based on mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S, i.e. ANLYF. Linear ANLYF-Gn is ANLYF with the N- and C-terminal extensions of Ala-(Gly)n-3 and -(Gly)2-Cys-Ala residues, respectively, with n=6-8, and circular ANLYF-Gn is ANLYF with the C and N termini cross-linked by the linker of the -Gly-Gly-Ala-(Gly)n-3 sequence, n=6-8
additional information
competitive hydrogen to deuterium and hydrogen to tritium kinetic isotope effects (KIEs) on the second-order rate constant (kcat/Km) are measured for wild-type and mutant ecDHFR variants to assess the intrinsic KIE on the catalyzed hydride transfer
additional information
-
competitive hydrogen to deuterium and hydrogen to tritium kinetic isotope effects (KIEs) on the second-order rate constant (kcat/Km) are measured for wild-type and mutant ecDHFR variants to assess the intrinsic KIE on the catalyzed hydride transfer
additional information
-
preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase (TmDHFR) dimer interface increases the melting temperature to about 60°C, approximately 9°C higher than that measured for the Escherichia coli enzyme. The steady-state and pre-steady-state rate constants measured for Xet-3 were similar to those of dimeric TmDHFR but significantly lower than those of the parent dihydrofolate reductase from Escherichia coli
additional information
replacement of an integral alpha-helical segment of Escherichia coli DHFR with segments from a number of different organisms (Listeria grayi, Haemophilus influenzae, Streptococcus dysgalactiae, Saccharomyces cerevisiae, Plasmodium falciparum, Homo sapiens). The parameters most sensitive to helical replacement are KM and Ki. The rank orders of KM, Ki, change of Tm, and protein stability against unfolding to the molten-globule state are significantly correlated with helical content
additional information
-
replacement of an integral alpha-helical segment of Escherichia coli DHFR with segments from a number of different organisms (Listeria grayi, Haemophilus influenzae, Streptococcus dysgalactiae, Saccharomyces cerevisiae, Plasmodium falciparum, Homo sapiens). The parameters most sensitive to helical replacement are KM and Ki. The rank orders of KM, Ki, change of Tm, and protein stability against unfolding to the molten-globule state are significantly correlated with helical content
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
60
-
preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface increases the melting temperature to about 60°C, approximately 9°C higher than that measured for the Escherichia coli enzyme
37
-
with NADPH bound all mutants retain activity longer than the wild-type enzyme, half-life of the wild-type enzyme 32 days
37
-
without NADPH, 50% loss of activity of wild-type and mutant enzymes after 60 h
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
7,8-dihydrofolate and folate protect against ethoxyformic anhydride modification
-
are more mobile. Betweeen EcDHFR and TmDHFR there is a shift in melting temperature of 26 K
-
at ionic strengths below the intracellular ion concentration-derived ionic strength in Escherichia coli ( at or below 0.237 M), the DHFR M20 loop tends to adopt open/closed conformations, and rarely an occluded loop state. As the ionic strength exceeds the physiological ionic strength of 0.237 M, the loop tends to adopt a closed/occluded conformation. the solution ionic strength affects the M20 loop stability differently depending on its conformation: High ionic strengths stabilize the occluded conformation more than low ionic strengths, as Ca2+ ions can approach E17 of the M20 loop and stabilize its orientation, whereas they are distal from the E17 at low ionic strengths. Both low and high ionic strengths can stabilize the closed conformation,
but they do not significantly affect the stability of the open conformation
comparison of the temperature dependence of stability and of flexibility between Thermotoga maritima and Escherichia coli enzymes. The TmDHFR dimer is overall not significantly more rigid than EcDHFR monomer. The TmDHFR protein core and most residues at the dimer interface exhibit smaller fluctuations than in EcDHFR, regions that are opposite to the interface
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
wild-type and mutants from high expression vector pCOCK, large scale purification
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
a variant (Xet-3) of dihydrofolate reductase from Escherichia coli constructed by introducing residues located at the Thermotoga maritima dihydrofolate reductase (TmDHFR) dimer interface is expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli
expression in Escherichia coli
folA gene, into multicopy vector pBR322, gene enrichment by Mu-mediated transposition
-
high expression rate of wild-type and mutants from expression vector pCOCK
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
kinetic of refolding upon dilution of unfolded enzyme in 4.5 M urea to 1.29 M urea
-
refolding of enzyme reversibly unfolded in 7 M urea, effect of several peptide fragments, derived from limited proteolytic cleavage of dihydrofolate reductase on the attainment of the folded state
-
study on kinetic folding of urea-denatured dihydrofolate reductase and comparison with Haloferax volcanii enzyme. Folding follows similar kinetics for both enzymes, with a 5-ms stopped-flow burst-phase species that folds to the native state through two sequential intermediateswith relaxation times of 0.1-3 sec and 25-100 sec. The unfolding of Haloferax volcanii enzyme at low ionic strength is relatively slow. Increased KCl concentrations slow the urea-induced unfolding of both enzymes, but much less than expected from equilibrium studies. Unfolding rates are relatively independent of ionic strength
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
study of protein dynamics, using a pump-probe method that employs pulsed-laser photothermal heating of a gold nanoparticle (AuNP) to directly excite a local region of the protein structure and transient absorbance to probe the effect on enzyme activity. Activity is accelerated by pulsed-laser excitation when the AuNP is attached close to a network of coupled motions in DHFR. No rate acceleration is observed when the AuNP is attached away from the network with pulsed excitation, or for any attachment site with continuous wave excitation
drug development
medicine
-
enzyme is a target of several anti-folate inhibitory drugs to combat bacteria, protozoa and cancer
drug development
-
receptor-based pharmacophore models based on ensembles of protein conformations from molecular dynamics simulations of enzyme in complex with NADPH in both the closed and open conformation of the M20 loop. Optimal models identify enzyme inhibitors over druglike noninhibitors. Model performance improves with increased dynamic sampling
drug development
the enzyme represents an attractive target for inhibitor design to disrupt systems that require rapid DNA turnover, e.g. proliferating cancer cells and pathogenic microbes
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Kraut, J.; Matthews, D.A.
Dihydrofolate reductase
Biol. Macromol. and Assem. (Junak, F. , McPherson, eds. )
3
1-71
1987
Klebsiella aerogenes, Bacteria, Gallus gallus, Escherichia coli, Homo sapiens, Lacticaseibacillus casei, Mus musculus, Pigeon, protozoa, vertebrata
-
Freisheim, J.H.; Matthews, D.A.
The comparative biochemistry of dihydrofolate reductase
Folate Antagonists Ther. Agents (Sirotnak, F. M. , ed. )
1
69-131
1984
Bacteria, Tequatrovirus T4, Bos taurus, Gallus gallus, Crithidia fasciculata, Streptococcus pneumoniae, Escherichia coli, Enterococcus faecium, Homo sapiens, Lacticaseibacillus casei, Mammalia, Mus musculus, protozoa, Sus scrofa, vertebrata
-
Roth, B.; Bliss, E.; Beddell, C.R.
Inhibitors of dihydrofolate reductase
Top. Mol. Struct. Biol.
3
363-393
1983
Bos taurus, Gallus gallus, Escherichia coli, Enterococcus faecium, Homo sapiens, Lacticaseibacillus casei, Sus scrofa
-
Morrison, J.F.; Stone, S.R.
Mechanism of the reaction catalyzed by dihydrofolate reductase from Escherichia coli: pH and deuterium isotope effects with NADPH as the variable substrate
Biochemistry
27
5499-5506
1988
Escherichia coli
Hall, J.G.; Frieden, C.
Protein fragments as probes in the study of protein folding mechanisms: differential effects of dihydrofolate reductase fragments on the refolding of the intact protein
Proc. Natl. Acad. Sci. USA
86
3060-3064
1989
Escherichia coli
Frieden, C.
Refolding of Escherichia coli dihydrofolate reductase: sequential formation of substrate binding sites
Proc. Natl. Acad. Sci. USA
87
4413-4416
1990
Escherichia coli
Bolin, J.T.; Filman, D.J.; Matthews, D.A.; Hamlin, R.C.; Kraut, J.
Crystal structures of Escherichia coli and Lactobacillus casei dihydrofolate reductase refined at 1.7 A resolution. I. General features and binding of methotrexate
J. Biol. Chem.
257
13650-13662
1982
Escherichia coli, Lacticaseibacillus casei
Roth, B.
Design of dihydrofolate reductase inhibitors from X-ray crystal structures
Fed. Proc.
45
2765-2772
1986
Gallus gallus, Escherichia coli, Lacticaseibacillus casei
Then, R.L.
Purification of guanosine triphosphate cyclohydrolase I and dihydrofolate reductase on a dihydrofolate-Sepharose affinity column
Anal. Biochem.
100
122-128
1979
Escherichia coli, Rattus norvegicus, Escherichia coli B / ATCC 11303, Escherichia coli MB 1428 / B / ATCC 11303
Kaufman, B.T.
Methotrexate-agarose in the purification of dihydrofolate reductase
Methods Enzymol.
34
272-281
1974
Tequatrovirus T4, Bos taurus, Saccharomyces cerevisiae, Gallus gallus, Cricetulus sp., Escherichia coli, Lacticaseibacillus casei, Mus musculus, Rattus norvegicus, Salmonella enterica subsp. enterica serovar Typhimurium
Huennekens, F.M.; Vitols, K.S.; Whiteley, J.M.; Neef, V.G.
Dihydrofolate reductase
Methods Cancer Res.
13
199-225
1976
Tequatrovirus T4, Bos taurus, Gallus gallus, Cricetulus sp., Escherichia coli, Enterococcus faecalis, Enterococcus faecium, Lacticaseibacillus casei, Mammalia, Mus musculus, Sus scrofa
-
Stone, S.R.; Morrison, J.F.
Dihydrofolate reductase from Escherichia coli: the kinetic mechanism with NADPH and reduced acetylpyridine adenine dinucleotide phosphate as substrates
Biochemistry
27
5493-5499
1988
Escherichia coli
Poe, M.; Greenfield, N.J.; Hirshfield, J.M.; Williams, M.N.; Hoogsteen, K.
Dihydrofolate reductase. Purification and characterization of the enzyme from an amethopterin-resistant mutant of Escherichia coli
Biochemistry
11
1023-1030
1972
Escherichia coli, Escherichia coli MB 1428 / B / ATCC 11303
Baccanari, D.; Phillips, A.; Smith, S.; Sinski, D.; Burchall, J.
Purification and properties of Escherichia coli dihydrofolate reductase
Biochemistry
14
5267-5273
1975
Escherichia coli
Baccanari, D.P.; Averett, D.; Briggs, C.; Burchall, J.
Escherichia coli dihydrofolate reductase: isolation and characterization of two isozymes
Biochemistry
16
3566-3572
1977
Escherichia coli
Poe, M.; Breeze, A.S.; Wu, J.K.; Short, C.R.; Hoogsteen, K.
Dihydrofolate reductase from trimethoprim-resistant Escherichia coli MB 3746 and MB 3747. Purification, amino acid composition, and some kinetic properties
J. Biol. Chem.
254
1799-1805
1979
Escherichia coli
Erickson, J.S.; Mathews, C.K.
Dihydrofolate reductases of Escherichia coli and bacteriophage Tr. A spectrofluorometric study
Biochemistry
12
372-380
1973
Tequatrovirus T4, Escherichia coli, Escherichia coli MB 1428 / B / ATCC 11303
Smith, S.L.; Stone, D.; Novak, P.; Baccanari, D.P.; Burchall, J.J.
R plasmid dihydrofolate reductase with subunit structure
J. Biol. Chem.
254
6222-6225
1979
Escherichia coli
Rood, J.I.; Laird, A.J.; Williams, J.W.
Cloning of the Escherichia coli K-12 dihydrofolate reductase gene following mu-mediated transposition
Gene
8
255-265
1980
Escherichia coli
Boles, J.O.; Lewinski, K.; Kuncle, M.G.; Hatada, M.; Lebioda, L.; Dunlap, R.B.; Odom, J.D.
Expression, characterization and crystallographic analysis of telluromethionyl dihydrofolate reductase
Acta Crystallogr. Sect. D
51
731-739
1995
Escherichia coli (P0ABQ4), Escherichia coli
Shaw, D.; Odom, J.D.; Dunlap, R.B.
High expression and steady-state kinetic characterization of methionine site-directed mutants of Escherichia coli methionyl- and selenomethionyl-dihydrofolate reductase
Biochim. Biophys. Acta
1429
401-410
1999
Escherichia coli
Schnell, J.R.; Dyson, H.J.; Wright, P.E.
Structure, dynamics, and catalytic function of dihydrofolate reductase
Annu. Rev. Biophys. Biomol. Struct.
33
119-140
2004
Escherichia coli (P0ABQ4), Escherichia coli
Venkitakrishnan, R.P.; Zaborowski, E.; McElheny, D.; Benkovic, S.J.; Dyson, H.J.; Wright, P.E.
Conformational changes in the active site loops of dihydrofolate reductase during the catalytic cycle
Biochemistry
43
16046-16055
2004
Escherichia coli (P0ABQ4), Escherichia coli
Zolli-Juran, M.; Cechetto, J.D.; Hartlen, R.; Daigle, D.M.; Brown, E.D.
High throughput screening identifies novel inhibitors of Escherichia coli dihydrofolate reductase that are competitive with dihydrofolate
Bioorg. Med. Chem. Lett.
13
2493-2496
2003
Escherichia coli
Giladi, M.; Altman-Price, N.; Levin, I.; Levy, L.; Mevarech, M.
FolM, a new chromosomally encoded dihydrofolate reductase in Escherichia coli
J. Bacteriol.
185
7015-7018
2003
Escherichia coli
Pucciarelli, S.; Spina, M.; Montecchia, F.; Lupidi, G.; Eleuteri, A.M.; Fioretti, E.; Angeletti, M.
Peroxynitrite-mediated oxidation of the C85S/C152E mutant of dihydrofolate reductase from Escherichia coli: functional and structural effects
Arch. Biochem. Biophys.
434
221-231
2005
Escherichia coli
Swanwick, R.S.; Maglia, G.; Tey, L.H.; Allemann, R.K.
Coupling of protein motions and hydrogen transfer during catalysis by Escherichia coli dihydrofolate reductase
Biochem. J.
394
259-265
2006
Escherichia coli (P0ABQ4), Escherichia coli
Antikainen, N.M.; Smiley, R.D.; Benkovic, S.J.; Hammes, G.G.
Conformation coupled enzyme catalysis: single-molecule and transient kinetics investigation of dihydrofolate reductase
Biochemistry
44
16835-16843
2005
Escherichia coli (P0ABQ4)
Pu, J.; Ma, S.; Garcia-Viloca, M.; Gao, J.; Truhlar, D.G.; Kohen, A.
Nonperfect synchronization of reaction center rehybridization in the transition state of the hydride transfer catalyzed by dihydrofolate reductase
J. Am. Chem. Soc.
127
14879-14886
2005
Escherichia coli
Ohmae, E.; Fukumizu, Y.; Iwakura, M.; Gekko, K.
Effects of mutation at methionine-42 of Escherichia coli dihydrofolate reductase on stability and function: implication of hydrophobic interactions
J. Biochem.
137
643-652
2005
Escherichia coli
Iwakura, M.; Maki, K.; Takahashi, H.; Takenawa, T.; Yokota, A.; Katayanagi, K.; Kamiyama, T.; Gekko, K.
Evolutional design of a hyperactive cysteine- and methionine-free mutant of Escherichia coli dihydrofolate reductase
J. Biol. Chem.
281
13234-13246
2006
Escherichia coli (P0ABQ4), Escherichia coli
Summerfield, R.L.; Daigle, D.M.; Mayer, S.; Mallik, D.; Hughes, D.W.; Jackson, S.G.; Sulek, M.; Organ, M.G.; Brown, E.D.; Junop, M.S.
A 2.13 A structure of E. coli dihydrofolate reductase bound to a novel competitive inhibitor reveals a new binding surface involving the M20 loop region
J. Med. Chem.
49
6977-6986
2006
Escherichia coli (P0ABQ4), Escherichia coli
McElheny, D.; Schnell, J.R.; Lansing, J.C.; Dyson, H.J.; Wright, P.E.
Defining the role of active-site loop fluctuations in dihydrofolate reductase catalysis
Proc. Natl. Acad. Sci. USA
102
5032-5037
2005
Escherichia coli (P0ABQ4)
Wang, L.; Goodey, N.M.; Benkovic, S.J.; Kohen, A.
Coordinated effects of distal mutations on environmentally coupled tunneling in dihydrofolate reductase
Proc. Natl. Acad. Sci. USA
103
15753-15758
2006
Escherichia coli
Svensson, A.K.; Zitzewitz, J.A.; Matthews, C.R.; Smith, V.F.
The relationship between chain connectivity and domain stability in the equilibrium and kinetic folding mechanisms of dihydrofolate reductase from E.coli
Protein Eng. Des. Sel.
19
175-185
2006
Escherichia coli (P0ABQ4), Escherichia coli
Boehr, D.D.; McElheny, D.; Dyson, H.J.; Wright, P.E.
The dynamic energy landscape of dihydrofolate reductase catalysis
Science
313
1638-1642
2006
Escherichia coli (P0ABQ4), Escherichia coli
Krahn, J.M.; Jackson, M.R.; DeRose, E.F.; Howell, E.E.; London, R.E.
Crystal structure of a type II dihydrofolate reductase catalytic ternary complex
Biochemistry
46
14878-14888
2007
Escherichia coli (P00383)
Feng, J.; Goswami, S.; Howell, E.E.
R67, the other dihydrofolate reductase: rational design of an alternate active site configuration
Biochemistry
47
555-565
2008
Escherichia coli
Boehr, D.D.; Dyson, H.J.; Wright, P.E.
Conformational relaxation following hydride transfer plays a limiting role in dihydrofolate reductase catalysis
Biochemistry
47
9227-9233
2008
Escherichia coli (P0ABQ4), Escherichia coli
Ohmae, E.; Tatsuta, M.; Abe, F.; Kato, C.; Tanaka, N.; Kunugi, S.; Gekko, K.
Effects of pressure on enzyme function of Escherichia coli dihydrofolate reductase
Biochim. Biophys. Acta
1784
1115-1121
2008
Escherichia coli (P0ABQ4), Escherichia coli
Meinhold, L.; Clement, D.; Tehei, M.; Daniel, R.; Finney, J.L.; Smith, J.C.
Protein dynamics and stability: the distribution of atomic fluctuations in thermophilic and mesophilic dihydrofolate reductase derived using elastic incoherent neutron scattering
Biophys. J.
94
4812-4818
2008
Geobacillus stearothermophilus, Escherichia coli (P0ABQ4)
Thielges, M.C.; Case, D.A.; Romesberg, F.E.
Carbon-deuterium bonds as probes of dihydrofolate reductase
J. Am. Chem. Soc.
130
6597-6603
2008
Escherichia coli
Takahashi, H.; Arai, M.; Takenawa, T.; Sota, H.; Xie, Q.H.; Iwakura, M.
Stabilization of hyperactive dihydrofolate reductase by cyanocysteine-mediated backbone cyclization
J. Biol. Chem.
282
9420-9429
2007
Escherichia coli (P0ABQ4), Escherichia coli
Chopra, S.; Dooling, R.M.; Horner, C.G.; Howell, E.E.
A balancing act between net uptake of water during dihydrofolate binding and net release of water upon NADPH binding in R67 dihydrofolate reductase
J. Biol. Chem.
283
4690-4698
2008
Escherichia coli (P00383), Escherichia coli
Lerner, M.G.; Bowman, A.L.; Carlson, H.A.
Incorporating dynamics in E. coli dihydrofolate reductase enhances structure-based drug discovery
J. Chem. Inf. Model.
47
2358-2365
2007
Escherichia coli
Arai, M.; Kondrashkina, E.; Kayatekin, C.; Matthews, C.R.; Iwakura, M.; Bilsel, O.
Microsecond hydrophobic collapse in the folding of Escherichia coli dihydrofolate reductase, an alpha/beta-type protein
J. Mol. Biol.
368
219-229
2007
Escherichia coli (P0ABQ4), Escherichia coli
Chen, J.; Dima, R.I.; Thirumalai, D.
Allosteric communication in dihydrofolate reductase: signaling network and pathways for closed to occluded transition and back
J. Mol. Biol.
374
250-266
2007
Escherichia coli (P0ABQ4), Escherichia coli
Gloss, L.M.; Topping, T.B.; Binder, A.K.; Lohman, J.R.
Kinetic folding of Haloferax volcanii and Escherichia coli dihydrofolate reductases: haloadaptation by unfolded state destabilization at high ionic strength
J. Mol. Biol.
376
1451-1462
2008
Escherichia coli, Haloferax volcanii
Divya, N.; Grifith, E.; Narayana, N.
Structure of the Q67H mutant of R67 dihydrofolate reductase-NADP+ complex reveals a novel cofactor binding mode
Protein Sci.
16
1063-1068
2007
Escherichia coli (P00383)
Khavrutskii, I.V.; Price, D.J.; Lee, J.; Brooks, C.L.
Conformational change of the methionine 20 loop of Escherichia coli dihydrofolate reductase modulates pKa of the bound dihydrofolate
Protein Sci.
16
1087-1100
2007
Escherichia coli (P0ABQ4), Escherichia coli
Spina, M.; Cuccioloni, M.; Mozzicafreddo, M.; Montecchia, F.; Pucciarelli, S.; Eleuteri, A.M.; Fioretti, E.; Angeletti, M.
Mechanism of inhibition of wt-dihydrofolate reductase from E. coli by tea epigallocatechin-gallate
Proteins
72
240-251
2008
Escherichia coli
Gangjee, A.; Qiu, Y.; Li, W.; Kisliuk, R.L.
Potent dual thymidylate synthase and dihydrofolate reductase inhibitors: classical and nonclassical 2-amino-4-oxo-5-arylthio-substituted-6-methylthieno[2,3-d]pyrimidine antifolates
J. Med. Chem.
51
5789-5797
2008
Escherichia coli, Toxoplasma gondii, Homo sapiens (P00374), Homo sapiens
Groff, D.; Thielges, M.C.; Cellitti, S.; Schultz, P.G.; Romesberg, F.E.
Efforts toward the direct experimental characterization of enzyme microenvironments: tyrosine100 in dihydrofolate reductase
Angew. Chem. Int. Ed. Engl.
48
3478-3481
2009
Escherichia coli (P0ABQ4)
Banjanac, M.; Tatic, I.; Ivezic, Z.; Tomic, S.; Dumic, J.
Pyrimido-pyrimidines: A novel class of dihydrofolate reductase inhibitors
Food Technol. Biotechnol.
47
236-245
2009
Homo sapiens (P00374), Escherichia coli (P0ABQ4)
-
Gangjee, A.; Li, W.; Kisliuk, R.L.; Cody, V.; Pace, J.; Piraino, J.; Makin, J.
Design, synthesis, and X-ray crystal structure of classical and nonclassical 2-amino-4-oxo-5-substituted-6-ethylthieno[2,3-d]pyrimidines as dual thymidylate synthase and dihydrofolate reductase inhibitors and as potential antitumor agents
J. Med. Chem.
52
4892-4902
2009
Escherichia coli, Toxoplasma gondii, Homo sapiens (P00374), Homo sapiens
Batruch, I.; Javasky, E.; Brown, E.D.; Organ, M.G.; Johnson, P.E.
Thermodynamic and NMR analysis of inhibitor binding to dihydrofolate reductase
Bioorg. Med. Chem.
18
8485-8492
2010
Escherichia coli (P0ABQ4), Escherichia coli
Murakami, C.; Ohmae, E.; Tate, S.; Gekko, K.; Nakasone, K.; Kato, C.
Cloning and characterization of dihydrofolate reductases from deep-sea bacteria
J. Biochem.
147
591-599
2010
Escherichia coli, Photobacterium profundum (D1MX70), Photobacterium profundum, Moritella yayanosii (D1MX71), Moritella yayanosii, Moritella japonica (D1MX72), Moritella japonica, Shewanella violacea (D1MYR1), Shewanella violacea
Yokota, A.; Takahashi, H.; Takenawa, T.; Arai, M.
Probing the roles of conserved arginine-44 of Escherichia coli dihydrofolate reductase in its function and stability by systematic sequence perturbation analysis
Biochem. Biophys. Res. Commun.
391
1703-1707
2010
Escherichia coli
Kamath, G.; Howell, E.; Agarwal, P.
The tail wagging the dog: Insights into catalysis in R67 dihydrofolate reductase
Biochemistry
49
9078-9088
2010
Escherichia coli (P00383)
Liu, C.T.; Wang, L.; Goodey, N.M.; Hanoian, P.; Benkovic, S.J.
Temporally overlapped but uncoupled motions in dihydrofolate reductase catalysis
Biochemistry
52
5332-5334
2013
Escherichia coli
Horiuchi, Y.; Ohmae, E.; Tate, S.; Gekko, K.
Coupling effects of distal loops on structural stability and enzymatic activity of Escherichia coli dihydrofolate reductase revealed by deletion mutants
Biochim. Biophys. Acta
1804
846-855
2010
Escherichia coli (P0ABQ4), Escherichia coli
Stojkovic, V.; Perissinotti, L.L.; Willmer, D.; Benkovic, S.J.; Kohen, A.
Effects of the donor-acceptor distance and dynamics on hydride tunneling in the dihydrofolate reductase catalyzed reaction
J. Am. Chem. Soc.
134
1738-1745
2012
Escherichia coli (P0ABQ4), Escherichia coli
Srinivasan, B.; Tonddast-Navaei, S.; Skolnick, J.
Ligand binding studies, preliminary structure-activity relationship and detailed mechanistic characterization of 1-phenyl-6,6-dimethyl-1,3,5-triazine-2,4-diamine derivatives as inhibitors of Escherichia coli dihydrofolate reductase
Eur. J. Med. Chem.
103
600-614
2015
Escherichia coli (P0ABQ4), Escherichia coli
Srinivasan, B.; Skolnick, J.
Insights into the slow-onset tight-binding inhibition of Escherichia coli dihydrofolate reductase detailed mechanistic characterization of pyrrolo [3,2-f] quinazoline-1,3-diamine and its derivatives as novel tight-binding inhibitors
FEBS J.
282
1922-1938
2015
Escherichia coli (P0ABQ4)
Liu, C.T.; Francis, K.; Layfield, J.P.; Huang, X.; Hammes-Schiffer, S.; Kohen, A.; Benkovic, S.J.
Escherichia coli dihydrofolate reductase catalyzed proton and hydride transfers temporal order and the roles of Asp27 and Tyr100
Proc. Natl. Acad. Sci. USA
111
18231-18236
2014
Escherichia coli (P0ABQ4), Escherichia coli
Guo, J.; Loveridge, E.J.; Luk, L.Y.; Allemann, R.K.
Effect of dimerization on dihydrofolate reductase catalysis
Biochemistry
52
3881-3887
2013
Escherichia coli, Thermotoga maritima (Q60034), Thermotoga maritima, Thermotoga maritima DSM 3109 (Q60034)
Sah, S.; Shah, R.; Govindan, A.; Varada, R.; Rex, K.; Varshney, U.
Utilisation of 10-formyldihydrofolate as substrate by dihydrofolate reductase (DHFR) and 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) tranformylase/IMP cyclohydrolase (PurH) in Escherichia coli
Microbiology
164
982-991
2018
Escherichia coli (P0ABQ4), Escherichia coli, Escherichia coli K12 (P0ABQ4)
Wan, Q.; Bennett, B.C.; Wymore, T.; Li, Z.; Wilson, M.A.; Brooks, C.L.; Langan, P.; Kovalevsky, A.; Dealwis, C.G.
Capturing the catalytic proton of dihydrofolate reductase Implications for general acid-Base catalysis
ACS Catal.
11
5873-5884
2021
Escherichia coli (P0ABQ4), Escherichia coli
Kaur, S.; Bhattacharyya, R.; Banerjee, D.
Hydrochlorothiazide and indapamide bind the NADPH binding site of bacterial dihydrofolate reductase results of an in-silico study and their implications
In Silico Pharmacol.
8
005
2020
Escherichia coli (P0ABQ4)
Oyen, D.; Fenwick, R.B.; Aoto, P.C.; Stanfield, R.L.; Wilson, I.A.; Dyson, H.J.; Wright, P.E.
Defining the structural basis for allosteric product release from E. coli dihydrofolate reductase using NMR relaxation dispersion
J. Am. Chem. Soc.
139
11233-11240
2017
Escherichia coli (P0ABQ4), Escherichia coli
Babu, C.S.; Lim, C.
Influence of solution ionic strength on the stabilities of M20 loop conformations in apo E. coli dihydrofolate reductase
J. Chem. Phys.
154
195103
2021
Escherichia coli (P0ABQ4), Escherichia coli
Penhallurick, R.; Durnal, M.; Harold, A.; Ichiye, T.
Adaptations for pressure and temperature in dihydrofolate reductases
Microorganisms
9
1706
2021
Escherichia coli, Moritella yayanosii
Maffucci, I.; Laage, D.; Stirnemann, G.; Sterpone, F.
Differences in thermal structural changes and melting between mesophilic and thermophilic dihydrofolate reductase enzymes
Phys. Chem. Chem. Phys.
22
18361-18373
2020
Escherichia coli, Thermotoga maritima
Kozlowski, R.; Zhao, J.; Dyer, R.B.
Acceleration of catalysis in dihydrofolate reductase by transient, site-specific photothermal excitation
Proc. Natl. Acad. Sci. USA
118
e2014592118
2021
Escherichia coli (P0ABQ4)
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