Oxidant-induced Interprotein Disulfide Formation in Cardiac Protein DJ-1 Occurs via an Interaction with Peroxiredoxin 2

Abstract

The role and responses of the dimeric DJ-1 protein to cardiac oxidative stress is incompletely understood. H 2 O 2 induces a 50-kDa DJ-1 interprotein homodimer disulfide, known to form between Cys-53 on each subunit. A trimeric 75-kDa DJ-1 complex that mass spectrometry shows contained 2-Cys peroxiredoxin also formed and precedes the appearance of the disulfide dimer. These observations may represent peroxiredoxin sensing and transducing the oxidant signal to DJ-1. The dimeric disulfide DJ-1 complex was stabilized by auranofin, suggesting that thioredoxin recycles it in cells. Higher concentrations of H 2 O 2 concomitantly induce DJ-1 Cys-106 hyperoxidation (sulfination or sulfonation) in myocytes, perfused heart, or HEK cells. An oxidation-resistant C53A DJ-1 shows potentiated H 2 O 2 -induced Cys-106 hyperoxidation. DJ-1 also forms multiple disulfides with unknown target proteins during H 2 O 2 treatment, the formation of which is also potentiated in cells expressing the C53A mutant. This suggests that the intersubunit disulfide induces a conformational change that limits Cys-106 forming heterodisulfide protein complexes or from hyperoxidizing. High concentrations of H 2 O 2 also induce cell death, with DJ-1 Cys-106 sulfonation appearing causal in these events, as expression of C53A DJ-1 enhanced both Cys-106 sulfonation and cell death. Nonetheless, expression of the DJ-1 C106A mutant, which fully prevents hyperoxidation, also showed exacerbated cell death responses to H 2 O 2 . A rational explanation for these findings is that DJ-1 Cys-106 forms disulfides with target proteins to limit oxidantinduced cell death. However, when Cys-106 is hyperoxidized, formation of these potentially protective heterodimeric disulfide complexes is limited, and so cell death is exacerbated.

C Terminus of DJ-1 Determines Its Homodimerization, Deglycation Activity and Suppression of Ferroptosis

“…In addition, the results about the suppression of ferroptosis were consist with MGO detoxification ability as DJ-1 ΔC3 and L187E mutants lost the inhibition of ferroptosis compared with DJ-1 WT. Besides, the suppression was more obvious in DJ-1 ΔC3 and L187E mutants than V51C mutant, indicating that the hydrophobic C terminus may have other effects such as involving regulation of signaling pathway [4, 6, 38]. All these data supported our previous discovery that DJ-1 was a negative modulator of ferroptosis providing new opportunities to facilitate ferroptosis-based cancer therapy [14].…”

Oxidation of phenylalanyl-tRNA synthetase positively regulates translational quality control

“…These results are in contrast to the better characterized oxidation of a cysteine residue in ThrRS that alters proofreading activity (5). However, oxidation of proteins has been shown to generally impact the structure of proteins and encourage the formation of aggregates leading to altered cellular responses (15,19,22). Oxidative generation of an unstructured protein causes PheRS to function in a hyperaccurate manner by more rapidly hydrolyzing the ester linkage of noncognate Tyr-tRNA Phe both in vitro and in vivo.…”

“…Protein Oxidation Causes a Conformational Change of PheRS. Protein oxidation can lead to the formation of disulfide bonds and other amino acid changes that induce a conformational change (15,19). To investigate why editing-deficient and WT PheRS have different oxidation patterns, circular dichroism was used to explore the possibility of a conformational change in the editing-deficient PheRS.…”

The oxidized thiol proteome in aging and cataractous mouse and human lens revealed by ICAT labeling

“…Under oxidative stress, Cys106 will be sulfinated or sulfonated to shift to a more acidic isoform triggering an oxidation elimination response. The Cys46 and Cys53 have much less regulatory activity than Cys106 (Meulener et al ., ), but Cys53 was found to be playing an important role in the stabilization of PARK7, so the Cys106 can form heterodisulfide protein complexes (Fernandez‐Caggiano et al ., ). We did not detect Cys106 disulfide in PARK7, but dramatic oxidation was found in Cys46 and Cys53 implying that Cys106 may have been hyperoxidized due to PARK7 protein conformational change via extensive oxidation of Cys46 and Cys53 residues.…”