Oxidant-induced Interprotein Disulfide Formation in Cardiac Protein DJ-1 Occurs via an Interaction with Peroxiredoxin 2*
| Autor: | Manuel Mayr, Javier Barallobre-Barreiro, Hyun-Ju Cho, Mariana Fernández-Caggiano, Joseph R. Burgoyne, Philip Eaton, Ewald Schröder |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Male Proteomics Conformational change Immunoprecipitation Protein subunit Blotting Western Protein Deglycase DJ-1 Fluorescent Antibody Technique Peroxiredoxin 2 Biology Biochemistry 03 medical and health sciences 0302 clinical medicine Animals Humans Myocytes Cardiac Protein Interaction Domains and Motifs Disulfides Rats Wistar Molecular Biology Cells Cultured HEK 293 cells Cell Biology Hydrogen Peroxide Peroxiredoxins Oxidants Rats Oxidative Stress 030104 developmental biology HEK293 Cells Mutation Biophysics Thioredoxin Peroxiredoxin Microtubule-Associated Proteins Oxidation-Reduction 030217 neurology & neurosurgery |
| Popis: | The role and responses of the dimeric DJ-1 protein to cardiac oxidative stress is incompletely understood. H2O2 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 H2O2 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 H2O2-induced Cys-106 hyperoxidation. DJ-1 also forms multiple disulfides with unknown target proteins during H2O2 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 H2O2 also induce cell death, with DJ-1 Cys-106 sulfonation appearing causal in these events, as expressionof 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 H2O2. A rational explanation for these findings is that DJ-1 Cys-106 forms disulfides with target proteins to limit oxidant-induced cell death. However, when Cys-106 is hyperoxidized, formation of these potentially protective heterodimeric disulfide complexes is limited, and so cell death is exacerbated. |
| Databáze: | OpenAIRE |
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