Insight into the mechanism of an iron dioxygenase by resolution of steps following the FeIV=HO species.

Iron oxygenases generate elusive transient oxygen species to catalyze substrate oxygenation in a wide range of metabolic processes. Here we resolve the reaction sequence and structures of such intermediates for the archetypal non-heme Fe(II) and alpha-ketoglutarate-dependent dioxygenase TauD. Time-resolved Raman spectra of the initial species with (16)O(18)O oxygen unequivocally establish the Fe(IV) horizontal lineO structure. (1)H/(2)H substitution reveals direct interaction between the oxo group and the C1 proton of substrate taurine. Two new transient species were resolved following Fe(IV) horizontal lineO; one is assigned to the nu(FeO) mode of an Fe(III) horizontal line O(H) species, and a second is likely to arise from the vibration of a metal-coordinated oxygenated product, such as Fe(II) horizontal line O horizontal line C(1) or Fe(II) horizontal line OOCR. These results provide direct insight into the mechanism of substrate oxygenation and suggest an alternative to the hydroxyl radical rebinding paradigm.