title: Purification, crystallization and preliminary crystallographic analysis of mouse myo-inositol oxygenase.
abstract: Myo-inositol oxygenase (MIOX) catalyzes the novel oxidative cleavage of myo-inositol (MI) and its epimer D-chiro inositol (DCI) to D-glucuronate. MIOX utilizes an Fe(II)/Fe(III) binuclear iron centre for the dioxygen-dependent cleavage of the C1-C6 bond in MI. Despite its key role in inositol metabolism, the structural basis of its unique four-electron oxidation mechanism and its substrate specificity remain unknown. In order to answer these questions and to facilitate the use of this key enzyme for the development of new therapeutic strategies for diabetes, the mouse enzyme has been cloned, expressed in Escherichia coli, purified and crystallized from 4.4 M sodium formate. The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 44.87, b = 77.26, c = 84.84 angstroms, and diffract to 2.8 angstroms resolution.
authors.affiliation: Centre for Molecular Biodiscovery, University of Auckland, Private Bag 92019, Auckland, New Zealand. pbro100@ec.auckland.ac.nz
authors.forename: Peter M
authors.initials: PM
authors.lastname: Brown
authors.forename: Tom T
authors.initials: TT
authors.lastname: Caradoc-Davies
authors.forename: James M
authors.initials: JM
authors.lastname: Dickson
authors.forename: Garth J S
authors.initials: GJ
authors.lastname: Cooper
authors.forename: Kerry M
authors.initials: KM
authors.lastname: Loomes
authors.forename: Edward N
authors.initials: EN
authors.lastname: Baker
mesh_terms: D000818:Animals; D001483:Base Sequence; D002236:Carbohydrate Conformation; D003460:Crystallization; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D007294:Inositol / Q000737:chemistry; D050562:Inositol Oxygenase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D051379:Mice; D008958:Models, Molecular; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D014961:X-Ray Diffraction
chemical_list: D017931:DNA Primers; D011994:Recombinant Proteins; D007294:Inositol; D050562:Inositol Oxygenase