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Information on EC 1.5.1.3 - dihydrofolate reductase and Organism(s) Haloferax volcanii and UniProt Accession P15093

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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:
Haloferax volcanii
UNIPROT: P15093
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The taxonomic range for the selected organisms is: Haloferax volcanii
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
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DHFR type IIIC
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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
5,6,7,8-tetrahydrofolate + NADP+
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KCl
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dihydrofolate reductase hvDHFR1 and hvDHFR2 unfold at KCl concentrations below 0.5 M. Above 1 M, the KCl dependence of the dihydrofolate reductase activities can be attributed to the effect of salt on substrate affinity
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
trimethoprim
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hDHFR-1 is much more resistant than hDHFR-2
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KCl
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hDHFR-2, activation is maximal at 0.5 M
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.08
7,8-dihydrofolate
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lowest Km value in 3.0 M KCl, recombinant wild-type enzyme
additional information
additional information
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overview Km-values of wild-type and mutant enzymes decreasing with increasing concentration of KCl
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.14 - 100
7,8-dihydrofolate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
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isozyme hDHFR-1
6
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isozyme hDHFR-2
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
Manually annotated by BRENDA team
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
NMR-derived binary hvDHFR1:folate complex structure is presented to explore any possible conformational changes that occur upon binding folate
the crystal structure is determined at 2.6 A resolution. The structure is in the apo state, with an open conformation of the active-site gully
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A31K
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site-directed mutagenesis
L30K
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site-directed mutagenesis
L30K/A31K
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site-directed mutagenesis
additional information
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construction of hdrAgene, hdrB gene and double deletion mutants
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the enzyme retains secondary structure at monovalent salt concentrations as low as 0.5 M
isozyme hDHFR-1, loss of more than 80% activity after exposure to 0.2 M KCl for 24 h at 24°C
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isozyme hDHFR-2 loss of less than 10% activity after exposure to 40 mM KCl for 18 h at 52°C
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
urea
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dihydrofolate reductase hvDHFR1 is ,1–2 kcal/mol more stable to urea denaturation than dihydrofolate reductase hvDHFR2. NaCl, KCl, and CsCl enhance the stability to urea denaturation
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
isozyme hDHFR-2 from hdrB gene overexpressed in Escherichia coli
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large quantities of recombinant wild-type enzyme from Escherichia coli, purification of mutants L30K, A31K and double mutant L30K/A31K recombinant from Escherichia coli
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
overexpression of hdrB gene in Escherichia coli
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overexpression of wild-type and mutant enzymes in Escherichia coli
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RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
study on kinetic folding of urea-denaturedc dihydrofolate reductase and comparison with Escherichia coli 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
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very fast and total reconstitution of active recombinant wild-type enzyme from inclusion bodies due to overexpression in E. coli by 6 M guanidine hydrochloride followed by dilution into 1 M NaCl or KCl solution
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ortenberg, R.; Rozenblatt-Rosen, O.; Mevarech, M.
The extremely halophilic archaeon Haloferax volcanii has two very different dihydrofolate reductases
Mol. Microbiol.
35
1493-1505
2000
Haloferax volcanii
Manually annotated by BRENDA team
Blecher, O.; Goldman, S.; Mevarech, M.
High expression in Escherichia coli of the gene coding for dihydrofolate reductase of the extremely halophilic archaebacterium Haloferax volcanii. Reconstitution of the active enzyme and mutation studies
Eur. J. Biochem.
216
199-203
1993
Haloferax volcanii
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
Boroujerdi, A.F.; Young, J.K.
NMR-derived folate-bound structure of dihydrofolate reductase 1 from the halophile Haloferax volcanii
Biopolymers
91
140-114
2009
Haloferax volcanii (P15093), Haloferax volcanii, Haloferax volcanii DSM 3757 (P15093)
Manually annotated by BRENDA team
Wright, D.B.; Banks, D.D.; Lohman, J.R.; Hilsenbeck, J.L.; Gloss, L.M.
The effect of salts on the activity and stability of Escherichia coli and Haloferax volcanii dihydrofolate reductases
J. Mol. Biol.
323
327-344
2002
Haloferax volcanii
Manually annotated by BRENDA team
Pieper, U.; Kapadia, G.; Mevarech, M.; Herzberg, O.
Structural features of halophilicity derived from the crystal structure of dihydrofolate reductase from the Dead Sea halophilic archaeon, Haloferax volcanii
Structure
6
75-88
1998
Haloferax volcanii (P15093), Haloferax volcanii, Haloferax volcanii DSM 3757 (P15093)
Manually annotated by BRENDA team