Crystallization and preliminary characterization of dihydropteridine reductase from Dictyostelium discoideum.
Dihydropteridine reductase from Dictyostelium discoideum (dicDHPR) can produce D-threo-BH(4) [6R-(1'R,2'R)-5,6,7,8-tetrahydrobiopterin], a stereoisomer of L-erythro-BH(4), in the last step of tetrahydrobiopterin (BH(4)) recycling. In this reaction, DHPR uses NADH as a cofactor to reduce quinonoid dihydrobiopterin back to BH(4). To date, the enzyme has been purified to homogeneity from many sources. In this report, the dicDHPR-NAD complex has been crystallized using the hanging-drop vapour-diffusion method with PEG 3350 as a precipitant. Rectangular-shaped crystals were obtained. Crystals grew to maximum dimensions of 0.4 x 0.6 x 0.1 mm. The crystal belonged to space group P2(1), with unit-cell parameters a = 49.81, b = 129.90, c = 78.76 A, beta = 100.00 degrees , and contained four molecules in the asymmetric unit, forming two closely interacting dicDHPR-NAD dimers. Diffraction data were collected to 2.16 A resolution using synchrotron radiation. The crystal structure has been determined using the molecular-replacement method.
Chen
Cong
C
Division of Applied Life Science (BK21 Program), Gyeongsang National University, Jinju 660-701, Republic of Korea.
Seo
Kyung Hye
KH
Kim
Hye Lim
HL
Zhuang
Ningning
N
Park
Young Shik
YS
Lee
Kon Ho
KH
D000818:Animals; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray; D004023:Dictyostelium / Q000201:enzymology*; D004093:Dihydropteridine Reductase / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D015800:Protozoan Proteins / Q000737:chemistry* / Q000302:isolation & purification
D015800:Protozoan Proteins; D004093:Dihydropteridine Reductase