How Does Single Oxygen Atom Addition Affect the Properties of an Fe−Nitrile Hydratase Analogue? The Compensatory Role of the Unmodified Thiolate

Nitrile hydratase (NHase) is one of a growing number of enzymes shown to contain post-translationally modified cysteine sulfenic acids (Cys-SOH). Cysteine sulfenic acids have been shown to
play diverse roles in cellular processes, including transcriptional regulation, signal transduction, and the
regulation of oxygen metabolism and oxidative stress responses. The function of the cysteine sulfenic acid
coordinated to the iron active site of NHase is unknown. Herein we report the first example of a sulfenate-ligated iron complex, [FeIII(ADIT)(ADIT-O)]+ (5), and compare its electronic and magnetic properties with
those of structurally related complexes in which the sulfur oxidation state and protonation state have been
systematically altered. Oxygen atom addition was found to decrease the unmodified thiolate Fe−S bond
length and blue-shift the ligand-to-metal charge-transfer band (without loss of intensity). S K-edge X-ray
absorption spectroscopy and density functional theory calculations show that, although the modified RS-O- fragment is incapable of forming a π bond with the FeIII center, the unmodified thiolate compensates for
this loss of π bonding by increasing its covalent bond strength. The redox potential shifts only slightly (75
mV), and the magnetic properties are not affected (the S = 1/2 spin state is maintained). The coordinated
sulfenate S−O bond is activated and fairly polarized (S+−O-). Addition of strong acids at low temperatures
results in the reversible protonation of sulfenate-ligated 5. An X-ray structure demonstrates that Zn2+ binds
to the sulfenate oxygen to afford [FeIII(ADIT)(ADIT-O-ZnCl3)] (6). The coordination of ZnCl3- to the RS-O-
unit causes the covalent overlap with the unmodified thiolate to increase further. A possible catalytic role
for the unmodified NHase thiolate, involving its ability to “tune” the electronics in response to protonation
of the sulfenate (RS-O-) oxygen and/or substrate binding, is discussed.