Demonstration by2H ENDOR Spectroscopy thatmyo-Inositol Binds via an Alkoxide Bridge to the Mixed-Valent Diiron Center ofmyo-Inositol Oxygenase

myo-Inositol oxygenase (MIOX) is a non-heme diiron oxygenase that cleaves cyclohexane-(1,2,3,5/4,6-hexa)-ol (myo-inositol, MI) to d-glucuronate. Here, we use 2H ENDOR spectroscopy to demonstrate that MI binds to the diiron(II/III) cofactor of MIOX via an alkoxide bridge, most likely involving O1. Analysis shows that MI adopts a symmetrical geometry in which the O−C−2H plane of the bridge is approximately orthogonal to the Fe−O−Fe plane.