title: Crystallization and preliminary crystallographic analysis of 2-aminophenol 1,6-dioxygenase complexed with substrate and with an inhibitor.
abstract: Dioxygen activation implemented by nonhaem Fe(II) enzymes containing the 2-His-1-carboxylate facial triad has been extensively studied in recent years. Extradiol dioxygenase is the archetypal member of this superfamily and catalyzes the oxygenolytic ring opening of catechol analogues. Here, the crystallization and preliminary X-ray analysis of 2-aminophenol 1,6-dioxygenase, an enzyme representing a minor subset of extradiol dioxygenases that catalyze the fission of 2-aminophenol rather than catecholic compounds, is reported. Crystals of the holoenzyme with FeII and of complexes with the substrate 2-aminophenol and the suicide inhibitor 4-nitrocatechol were grown using the cocrystallization method under the same conditions as used for the crystallization of the apoenzyme. The crystals belonged to space group C2 and diffracted to 2.3-2.7 Å resolution; the crystal that diffracted to the highest resolution had unit-cell parameters a=270.24, b=48.39, c=108.55 Å, β=109.57°. All X-ray data sets collected from diffraction-quality crystals were suitable for structure determination.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
authors.forename: De-Feng
authors.initials: DF
authors.lastname: Li
authors.forename: Jia-Yue
authors.initials: JY
authors.lastname: Zhang
authors.forename: Yanjie
authors.initials: Y
authors.lastname: Hou
authors.forename: Lei
authors.initials: L
authors.lastname: Liu
authors.forename: Shuang-Jiang
authors.initials: SJ
authors.lastname: Liu
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
mesh_terms: D000627:Aminophenols / Q000737:chemistry*; D001051:Apoenzymes / Q000737:chemistry / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002396:Catechols / Q000737:chemistry*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D020604:Comamonas testosteroni / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D049308:Dioxygenases / Q000737:chemistry* / Q000302:isolation & purification; D004791:Enzyme Inhibitors / Q000737:chemistry
chemical_list: D000627:Aminophenols; D001051:Apoenzymes; D001426:Bacterial Proteins; D002396:Catechols; D004791:Enzyme Inhibitors; C027667:2-aminophenol; C106307:2-aminophenol 1,6-dioxygenase; D049308:Dioxygenases; C001833:4-nitrocatechol