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Literature summary extracted from

  • Chen, C.; Seo, K.H.; Kim, H.L.; Zhuang, N.; Park, Y.S.; Lee, K.H.
    Crystallization and preliminary characterization of dihydropteridine reductase from Dictyostelium discoideum (2008), Acta Crystallogr. Sect. F, 64, 1013-1015.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.34 dicDHPR gene (residues 1-231) cloned into pET-28b expression plasmid with hexahistidine tag and thrombin cleavage site, expression in Escherichia coli BL21 (DE3) Dictyostelium discoideum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.5.1.34 dicDHPR-NAD complex by incubation in 20 mM Tris-HCl, pH 8.0, hanging-drop vapor diffusion (4 microl drops), 18°C (291 K), screening solution (0.1 M bis-tris, pH 6.5, 17% PEG 3350), hexahistidine tag not removed, x-ray diffraction Dictyostelium discoideum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.5.1.34 24600
-
 calculated, monomer Dictyostelium discoideum
1.5.1.34 25000
-
 SDS-PAGE, monomer Dictyostelium discoideum
1.5.1.34 50000
-
 gel-filtration chromatography (Superdex 200 column), dimer Dictyostelium discoideum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.5.1.34 quinonoid dihydrobiopterin + NADH + H+ Dictyostelium discoideum
-
 6R-(1'R,2'R)-5,6,7,8-tetrahydrobiopterin + NAD+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.34 Dictyostelium discoideum
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.1.34 cells centrifuged, washed with PBS (pH 7.4), resuspended in lysis buffer (50 mM Tris-HCl, pH 7.5), disrupted by sonication, centrifugation, supernatant filtered (0.45 microm), applied to a column with nickel-NTA beads, eluted (50 mM Tris-HCl, pH 7.5), active fractions pooled, concentrated, and exchanged into 50 mM sodium phosphate, pH 6.8, ultrafiltration, applied to a cation-exchange chromatography HS20 column, eluted with salt gradient at pH 6.8, concentration by ultrafiltration, gel-filtration chromatography with Superdex 200 column in 50 mM Tris-HCl, pH 8.0, concentration of eluted fractions in 20 mM Tris-HCl, pH 8.0, by ultrafiltration Dictyostelium discoideum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.34 quinonoid dihydrobiopterin + NADH + H+
-
 Dictyostelium discoideum 6R-(1'R,2'R)-5,6,7,8-tetrahydrobiopterin + NAD+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.5.1.34 dimer 2 * 25000, four molecules form 2 dimers in the asymmetric unit in crystals Dictyostelium discoideum

Synonyms

EC Number Synonyms Comment Organism
1.5.1.34 dicDHPR
-
 Dictyostelium discoideum
1.5.1.34 dihydropteridine reductase
-
 Dictyostelium discoideum

General Information

EC Number General Information Comment Organism
1.5.1.34 metabolism enzyme catalyzes the last step of tetrahydrobiopterin recycling Dictyostelium discoideum