Purification, crystallization and crystallographic analysis of Dictyostelium discoideum phenylalanine hydroxylase in complex with dihydrobiopterin and FeIII.
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.
Zhuang
Ningning
N
Division of Applied Life Science (BK21 Program), Gyeongsang National University, Jinju 660-701, Republic of Korea.
Seo
Kyung Hey
KH
Chen
Cong
C
Kim
Hye Lim
HL
Park
Young Shik
YS
Lee
Kon Ho
KH
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
D005290:Ferric Compounds; D001708:Biopterins; C017226:7,8-dihydrobiopterin; D010651:Phenylalanine Hydroxylase
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