Literature summary for 1.5.1.34 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.

Cloned(Commentary)

Cloned (Comment)	Organism
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)

Crystallization (Comment)	Organism
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]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Dictyostelium discoideum	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
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

Purification (Comment)

Organism

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)

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

Subunits

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

Synonyms

Synonyms	Comment	Organism
dicDHPR	-	Dictyostelium discoideum
dihydropteridine reductase	-	Dictyostelium discoideum

General Information

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