title: Purification, crystallization and crystallographic analysis of Dictyostelium discoideum phenylalanine hydroxylase in complex with dihydrobiopterin and FeIII.
abstract: Dictyostelium discoideum phenylalanine hydroxylase (DicPAH; residues 1-415) was expressed in Escherichia coli and purified for structural analysis. Apo DicPAH and DicPAH complexed with dihydrobiopterin (BH(2)) and Fe(III) were crystallized using 0.06 M PIPES pH 7.0, 26%(w/v) PEG 2000 by the hanging-drop vapour-diffusion method. Crystals of apo DicPAH and the DicPAH-BH(2)-Fe(III) complex diffracted to 2.6 and 2.07 A resolution, respectively, and belonged to space group P2(1), with unit-cell parameters a = 70.02, b = 85.43, c = 74.86 A, beta = 110.12 degrees and a = 70.97, b = 85.33, c = 74.89 A, beta = 110.23 degrees , respectively. There were two molecules in the asymmetric unit. The structure of DicPAH has been solved by molecular replacement.
authors.affiliation: Division of Applied Life Science (BK21 Program), Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Ningning
authors.initials: N
authors.lastname: Zhuang
authors.forename: Kyung Hey
authors.initials: KH
authors.lastname: Seo
authors.forename: Cong
authors.initials: C
authors.lastname: Chen
authors.forename: Hye Lim
authors.initials: HL
authors.lastname: Kim
authors.forename: Young Shik
authors.initials: YS
authors.lastname: Park
authors.forename: Kon Ho
authors.initials: KH
authors.lastname: Lee
mesh_terms: D001708:Biopterins / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D004023:Dictyostelium / Q000201:enzymology*; D005290:Ferric Compounds / Q000737:chemistry* / Q000378:metabolism; D010651:Phenylalanine Hydroxylase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding
chemical_list: D005290:Ferric Compounds; D001708:Biopterins; C017226:7,8-dihydrobiopterin; D010651:Phenylalanine Hydroxylase
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