Relationships between the Reversible Oxidation of the Single Cysteine Residue and the Physiological Function of the Mitochondrial Glutaredoxin S15 from .

Glutaredoxins (GRXs) are widespread proteins catalyzing deglutathionylation or glutathionylation reactions or serving for iron-sulfur (Fe-S) protein maturation. Previous studies highlighted a role of the mitochondrial class II GRXS15 in Fe-S cluster assembly, whereas only a weak glutathione-dependent oxidation activity was detected with the non-physiological roGFP2 substrate in vitro. Still, the protein must exist in a reduced form for both redox and Fe-S cluster binding functions. Therefore, this study aimed at examining the redox properties of GRXS15. The acidic pKa of the sole cysteine present in GRXS15 indicates that it should be almost totally under a thiolate form at mitochondrial pH and thus possibly subject to oxidation. Oxidizing treatments revealed that this cysteine reacts with HO or with oxidized glutathione forms. This leads to the formation of disulfide-bridge dimers and glutathionylated monomers which have redox midpoint potentials of -304 mV and -280 mV, respectively. Both oxidized forms are reduced by glutathione and mitochondrial thioredoxins. In conclusion, it appears that GRXS15 is prone to oxidation, forming reversible oxidation forms that may be seen either as a catalytic intermediate of the oxidoreductase activity and/or as a protective mechanism preventing irreversible oxidation and allowing Fe-S cluster binding upon reduction.