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Information on EC 1.5.1.3 - dihydrofolate reductase and Organism(s) Staphylococcus aureus and UniProt Accession P0A017

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EC Tree
     1 Oxidoreductases
         1.5 Acting on the CH-NH group of donors
             1.5.1 With NAD+ or NADP+ as acceptor
                1.5.1.3 dihydrofolate reductase
IUBMB Comments
The enzyme from animals and some micro-organisms also slowly reduces folate to 5,6,7,8-tetrahydrofolate.
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This record set is specific for:
Staphylococcus aureus
UNIPROT: P0A017
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Word Map
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The taxonomic range for the selected organisms is: Staphylococcus aureus
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
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7,8-dihydrofolate reductase
-
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dehydrogenase, tetrahydrofolate
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DHFR type IIIC
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dihydrofolate reductase
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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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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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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
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oxidation
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reduction
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PATHWAY SOURCE
PATHWAYS
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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 hide
9002-03-3
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
show the reaction diagram
the conserved residues Leu28, Val31, Ile50 and Leu54 forming a hydrophobic pocket in the enzyme are the binding site for the p-aminobenzamide moiety of the substrate dihydrofolate
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-
?
7,8-dihydrofolate + NADPH
5,6,7,8-tetrahydrofolate + NADP+
show the reaction diagram
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-
-
-
?
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
show the reaction diagram
dihydrofolate + NADPH + H+
tetrahydrofolate + NADP+
show the reaction diagram
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-
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
7,8-dihydrofolate + NADPH + H+
5,6,7,8-tetrahydrofolate + NADP+
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
AR-101
i.e. 5-[(2R)-2-cyclopropyl-7,8-dimethoxy-2H-chromene-5-ylmethyl]pyrimidine-2,4-diamine, binding structure from crystal structure, the cyclopropyl-dimethoxychromene moiety interacts with the conserved residues Leu28, Val31, Ile50 and Leu54 forming a hydrophobic pocket in the enzyme
Ar-102
i.e. 5-[(2S)-2-cyclopropyl-7,8-dimethoxy-2H-chromene-5-ylmethyl]pyrimidine-2,4-diamine, binding structure from crystal structure, the cyclopropyl-dimethoxychromene moiety interacts with the conserved residues Leu28, Val31, Ile50 and Leu54 forming a hydrophobic pocket in the enzyme
iclaprim
binding structure from crystal structure
(E)-1-(1-cyclopropyl-6,7-dimethoxyphthalazin-2(1H)-yl)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)prop-2-en-1-one
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(E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(1-(4-methoxyphenyl)phthalazin-2(1H)-yl)prop-2-en-1-one
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-
(E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(1-phenylphthalazin-2(1H)-yl)prop-2-en-1-one
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-
(E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(1-propylphthalazin-2(1H)-yl)prop-2-en-1-one
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(E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(6,7-dimethoxy-1-propylphthalazin-2(1H)-yl)prop-2-en-1-one
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(E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(6,7-dimethyl-1-propylphthalazin-2(1H)-yl)prop-2-en-1-one
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3-(2,4-diamino-6-methylpyrimidin-5-yl)-1-(3,4,5-trimethoxyphenyl)prop-2-yn-1-ol
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5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-ethylpyrimidine-2,4-diamine
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5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
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5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-propylpyrimidine-2,4-diamine
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5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]pyrimidine-2,4-diamine
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5-[3-(2-methoxy-2',6'-dimethylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
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5-[3-(2-methoxy-2'-methylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
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5-[3-(2-methoxybiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
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5-[3-(5-methoxy-2',6'-dimethylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
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5-[3-(5-methoxy-2'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
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5-[3-(5-methoxy-4'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
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5-[3-(5-methoxybiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
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5-[3-methoxy-3-(3,4,5-trimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
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6-methyl-5-[3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine
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6-methyl-5-[3-(3,4,5-trimethoxyphenyl)pent-1-yn-1-yl]pyrimidine-2,4-diamine
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6-methyl-5-[3-methyl-3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine
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BAL0030543
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a dihydrophthalazine inhibitor with antibacterial activity, overview
BAL0030544
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a dihydrophthalazine inhibitor with antibacterial activity, overview
BAL0030545
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a dihydrophthalazine inhibitor with antibacterial activity, overview
grape seed extract
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grape seed extract inhibits the dihydrofolate reductase activity and growth of Staphylococcus aureus. The grape seed extract-induced growth inhibition is reversed by adding, tetrahydrofolate, 5,10-methylenetetrahydrofolate or methionine to the medium
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trimethoprim
additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00268
7,8-dihydrofolate
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0.00238
NADPH
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00000023 - 0.0000009
AR-101
0.00000022 - 0.0000035
Ar-102
0.0000002 - 0.0000026
iclaprim
0.000006 - 0.0001
trimethoprim
0.0000007
(E)-1-(1-cyclopropyl-6,7-dimethoxyphthalazin-2(1H)-yl)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)prop-2-en-1-one
30°C, pH not specified in the publication
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0.0000012
(E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(1-(4-methoxyphenyl)phthalazin-2(1H)-yl)prop-2-en-1-one, (E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(1-phenylphthalazin-2(1H)-yl)prop-2-en-1-one
30°C, pH not specified in the publication
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0.0000012
(E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(1-phenylphthalazin-2(1H)-yl)prop-2-en-1-one
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30°C, pH not specified in the publication
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0.0000012
(E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(1-propylphthalazin-2(1H)-yl)prop-2-en-1-one
30°C, pH not specified in the publication
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0.0000011
(E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(6,7-dimethoxy-1-propylphthalazin-2(1H)-yl)prop-2-en-1-one
30°C, pH not specified in the publication
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0.0000012
(E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(6,7-dimethyl-1-propylphthalazin-2(1H)-yl)prop-2-en-1-one
30°C, pH not specified in the publication
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IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0011 - 0.0041
3-(2,4-diamino-6-methylpyrimidin-5-yl)-1-(3,4,5-trimethoxyphenyl)prop-2-yn-1-ol
0.000068 - 0.00067
5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-ethylpyrimidine-2,4-diamine
0.00027 - 0.0013
5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
0.000048 - 0.00055
5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-propylpyrimidine-2,4-diamine
0.00027 - 0.0069
5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]pyrimidine-2,4-diamine
0.00041 - 0.00613
5-[3-(2-methoxy-2',6'-dimethylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
0.0005 - 0.00142
5-[3-(2-methoxy-2'-methylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
0.00017 - 0.00077
5-[3-(2-methoxybiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
0.000073 - 0.00018
5-[3-(5-methoxy-2',6'-dimethylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
0.000073 - 0.00019
5-[3-(5-methoxy-2'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
0.00041 - 0.00096
5-[3-(5-methoxy-4'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
0.000042 - 0.00019
5-[3-(5-methoxybiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
0.00073 - 0.012
5-[3-methoxy-3-(3,4,5-trimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
0.00017 - 0.0012
6-methyl-5-[3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine
0.00012 - 0.0039
6-methyl-5-[3-(3,4,5-trimethoxyphenyl)pent-1-yn-1-yl]pyrimidine-2,4-diamine
0.000064 - 0.00075
6-methyl-5-[3-methyl-3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine
0.000023 - 0.0017
trimethoprim
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
DHFR is a critical enzyme in the maintenance of reduced folate pools used in the biosynthetic pathways of purines, thymidylate, methionine, glycine, pantothenic acid, and N-formyl-methionyl tRNA
physiological function
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
DYR_STAAU
159
0
18251
Swiss-Prot
-
PDB
SCOP
CATH
UNIPROT
ORGANISM
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
wild-type enzyme or mutant F98Y in ternary complex with cofactor NADPH and inhibitors AR-101, AR-102, or iclaprim, hanging-drop vapour-diffusion technique at 24°C, mixing of 30 mg/ml protein with 5 mM NADPH and 0.3 mg/ml inhibitor in 25% PEG 3350, 200 mM NaCl, and 100 mM bis-tris, pH 5.5, X-ray diffraction structure determination and analysis at 2.1-2.5 A resolution
crystal structure of the wild-type chromosomal DHFR from Staphylococcus aureus in complex with NADPH and trimethoprim is determined to 1.95 A resolution. The enzyme maintains the conserved fold of DHFR observed in other species with the active site core formed by an eight-stranded beta-sheet and four alpha-helices surrounding the core
purified recombinant wild-type and mutant enzymes bound to cofactor NADPH and inhibitors 5, 8, 10, and 15, hanging-drop vaporization method, 12 mg/ml protein is incubated with 1 mM inhibitor and 2 mM NADPH on ice for 2 h, mixing of equal volumes of the protein:ligand:NADPH solution with an optimized crystallization solution consisting of 15% PEG 10000, 150 mM sodium acetate, 100 mM MES, pH 6.5, and 5% butyrlactone, 5-7 days X-ray diffraction structure determination and analysis at 1.69-2.35 A resolutions, overview
purified recombinant wild-type and mutant enzymes in complex with inhibitor trimethoprim, hanging drop vapour diffusion method, 30.6 mg/ml wild-type enzyme protein mixed with 1 mM NADPH and 1 mM trimethoprim and incubated on ice for 3 h, mixing the protein 1:1 with a reservoir solution containing 30 mM citric acid/40 mM bis-tris propane pH 6.4, 13.3% PEG 3350, and 16.7% PEG 6000 and incubating at 22°C, hexagonal rod crystals form in 1-2 weeks, X-ray diffraction structure determination and analysis at 3.0 and 1.95 A resolution, respectively, molecular replacement and structure modelling
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F98Y
site-directed mutagenesis, the mutant enzyme shows reduced sensitivity to inhibitors compared to the wild-type enzyme, structure comparison to the wild-type, overview
N48E/N130D
site-directed mutagenesis, the S1 mutant enzyme shows improved expression levels and solubility. Inhibition kinetics and inhibitor binding thermodynamics in comparison to the wild-type enzyme, overview. In the absence of substrate and cofactor the active site of S1 DHFR is blocked, trimethoprim shows loss of potency and NADPH synergy on binding S1 DHFR
N48E/N130D/Y98F/A43G
site-directed mutagenesis, inhibition kinetics and inhibitor binding thermodynamics in comparison to the wild-type enzyme, overview
Y98F/A43G
inhibitor trimethoprim shows loss potency and NADPH synergy on binding S1 mutant DHFR. Mutation of residues Y98F/A43G in S1 mutant restores trimethoprim sensitivity and NADPH synergy
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-type and mutant enzmyes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 (AI) by adsorption chromatography and gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
expressed in Escherichia coli as a His-tagged fusion protein
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expression in Escherichia coli
expression of wild-type and mutant enzymes in Escherichia coli strain BL21 (AI)
overexpression of His-tagged wild-type and mutant enzmyes in Escherichia coli strain BL21(DE3) as soluble proteins
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
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significantly reduced inflammatory responses and mortality are observed in zebrafish infected with Staphylococcus aureus pre-incubated with grape seed extract. Grape seed extract might serve as an effective natural alternative for the control of food poisoning caused by Staphylococcus aureus with proper safety measure
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Sekiguchi, J.; Tharavichitkul, P.; Miyoshi-Akiyama, T.; Chupia, V.; Fujino, T.; Araake, M.; Irie, A.; Morita, K.; Kuratsuji, T.; Kirikae, T.
Cloning and characterization of a novel trimethoprim-resistant dihydrofolate reductase from a nosocomial isolate of Staphylococcus aureus CM.S2 (IMCJ1454)
Antimicrob. Agents Chemother.
49
3948-3951
2005
Staphylococcus aureus, Staphylococcus aureus CM.S2
Manually annotated by BRENDA team
Oefner, C.; Parisi, S.; Schulz, H.; Lociuro, S.; Dale, G.E.
Inhibitory properties and X-ray crystallographic study of the binding of AR-101, AR-102 and iclaprim in ternary complexes with NADPH and dihydrofolate reductase from Staphylococcus aureus
Acta Crystallogr. Sect. D
65
751-757
2009
Staphylococcus aureus (P0A017), Staphylococcus aureus
Manually annotated by BRENDA team
Bowker, K.E.; Caspers, P.; Gaucher, B.; MacGowan, A.P.
In vitro activities of three new dihydrofolate reductase inhibitors against clinical isolates of Gram-positive bacteria
Antimicrob. Agents Chemother.
53
4949-4952
2009
Streptococcus pneumoniae, Staphylococcus aureus, Streptococcus mitis
Manually annotated by BRENDA team
Frey, K.M.; Liu, J.; Lombardo, M.N.; Bolstad, D.B.; Wright, D.L.; Anderson, A.C.
Crystal structures of wild-type and mutant methicillin-resistant Staphylococcus aureus dihydrofolate reductase reveal an alternate conformation of NADPH that may be linked to trimethoprim resistance
J. Mol. Biol.
387
1298-1308
2009
Staphylococcus aureus (P0A016), Staphylococcus aureus
Manually annotated by BRENDA team
Heaslet, H.; Harris, M.; Fahnoe, K.; Sarver, R.; Putz, H.; Chang, J.; Subramanyam, C.; Barreiro, G.; Miller, J.
Structural comparison of chromosomal and exogenous dihydrofolate reductase from Staphylococcus aureus in complex with the potent inhibitor trimethoprim
Proteins
76
706-717
2009
Staphylococcus aureus, Staphylococcus aureus (P13955)
Manually annotated by BRENDA team
Kao, T.T.; Tu, H.C.; Chang, W.N.; Chen, B.H.; Shi, Y.Y.; Chang, T.C.; Fu, T.F.
Grape seed extract inhibits the growth and pathogenicity of Staphylococcus aureus by interfering with dihydrofolate reductase activity and folate-mediated one-carbon metabolism
Int. J. Food Microbiol.
141
17-27
2010
Staphylococcus aureus
Manually annotated by BRENDA team
Muddala, N.; White, J.; Nammalwar, B.; Pratt, I.; Thomas, L.; Bunce, R.; Berlin, K.; Bourne, C.
Inhibitor design to target a unique feature in the folate pocket of Staphylococcus aureus dihydrofolate reductase
Eur. J. Med. Chem.
200
112412
2020
Staphylococcus aureus (A0A0M3KKX1), Staphylococcus aureus, Staphylococcus aureus ET3-1 (A0A0M3KKX1)
Manually annotated by BRENDA team