Open AccessPurification, crystallization and crystallographic analysis of Dictyostelium discoideum phenylalanine hydroxylase in complex with dihydrobiopterin and Fe III
Author(s) -
Zhuang Ningning,
Seo Kyung Hey,
Chen Cong,
Kim HyeLim,
Park Young Shik,
Lee Kon Ho
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110007220
Subject(s) - dictyostelium discoideum , crystallization , phenylalanine hydroxylase , phenylalanine , chemistry , crystallography , stereochemistry , biochemistry , organic chemistry , amino acid , gene
Dictyostelium discoideum phenylalanine hydroxylase ( Dic PAH; residues 1–415) was expressed in Escherichia coli and purified for structural analysis. Apo Dic PAH and Dic PAH 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 Dic PAH and the Dic PAH–BH 2 –Fe III complex diffracted to 2.6 and 2.07 Å resolution, respectively, and belonged to space group P 2 1 , with unit‐cell parameters a = 70.02, b = 85.43, c = 74.86 Å, β = 110.12° and a = 70.97, b = 85.33, c = 74.89 Å, β = 110.23°, respectively. There were two molecules in the asymmetric unit. The structure of Dic PAH has been solved by molecular replacement.