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 FeII/FeIII 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 P21212 1, with unit-cell parameters a = 44.87, b = 77.26, c = 84.84 Å, and diffract to 2.8 Å resolution. © 2006 International Union of Crystallography All rights reserved.