Active Site Structures of the Escherichia coli N‑Hydroxylaminopurine Resistance Molybdoenzyme YcbX

X-ray absorption near-edge structure (XANES) and extended X-ray absorption fine structure (EXAFS) data have been used to characterize the coordination environment for the catalytic Mo site of Escherichia coli YcbX in two different oxidation states. In the oxidized state, the Mo­(VI) ion is coordinated by two terminal oxo ligands, a thiolate S atom from cysteine, and two S donors from the bidentate pyranopterin ene-1,2-dithiolate (pyranopterin dithiolene). Upon reduction, it is the more basic equatorial oxo ligand that is protonated, with a Mo–Oeq bond distance that is best described as either a short Mo4+–OH2 bond or a long Mo4+–OH bond. Mechanistic implications for substrate reduction are discussed in light of these structural details.