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Catalase and estradiol inhibit mitochondrial protein S-glutathionylation

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Abstract

Regulation and downstream effects of mitochondrial protein S-glutathionylation in response to oxidative stress are poorly understood. The study aim was to determine whether anti-oxidants such as catalase and estradiol alter mitochondrial protein S-glutathionylation and in turn affect apoptosis following ultraviolet B (UV-B) light irradiation. HeLa cells were transduced with increasing amounts of adenovirus encoding catalase (Ad-Cat) and β-galactosidase (Ad-Lac Z) or pre-incubated with estradiol before induction of apoptosis by UV-B light exposure. Inhibition of mitochondrial protein S-glutathionylation was assessed using autoantibodies specific for the non-S-glutathionylated form of PDC-E2. The percentage of apoptotic cells following UV-B irradiation were not significantly different between mock cells (cells with no virus infection) and Ad-Cat and Ad-Lac Z infected cells at all viral doses (all p > 0.050). Autoantibody staining of non-S-glutathionylated PDC-E2 in apoptotic cells was three times greater in only Ad-Cat infected cells compared to only Ad-Lac Z infected cells (81.3 ± 16.7 vs 26 ± 7.2 %, respectively, p = 0.030). Similarly estradiol treatment (33 and 100 nM) also significantly increased PDC-E2 staining in apoptotic cells compared to non-treated cells (both p < 0.010). The percentage of apoptotic cells was not significantly different with any of the estradiol concentrations (all p > 0.100). The observed procaspase 12 cleavage following UV-B irradiation suggests that a mitochondrial-independent apoptotic pathway was activated. In conclusion, following an apoptotic stimulus, estradiol may inhibit mitochondrial protein S-glutathionylation without inhibiting apoptosis. This effect may play a role in ninefold greater prevalence of autoantibodies against PDC-E2 in women with primary biliary cirrhosis.

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Abbreviations

Ad-Cat:

Adenovirus encoding catalase

Ad-Lac Z:

Adenovirus encoding β-galactosidase

DAPI:

4′,6′-Diamidino-2-phenylindole

ECL:

Enhanced chemiluminescence

HRP:

Horse radish peroxidase

m.o.i.:

Multiplicity of infection

PBC:

Primary biliary cirrhosis

PDC-E2:

E2 subunit of the pyruvate dehydrogenase complex

r PDC-E2:

Recombinant PDC-E2

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

UV-B:

Ultraviolet B light

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Acknowledgments

These studies were supported by an NIH grant T32DK07832 (JAO), and the Artzt Family PBC Foundation (JAO). Microscopy was performed at the MSSM-Microscopy Shared Research Facility, supported, in part, with funding from NIH-NCI shared resources grant (1 R24 CA095823-01).

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Authors and Affiliations

  1. Department of Medicine, The Mount Sinai School of Medicine, Box 1133, New York, NY, 10029, USA

    Bin Hu, Jorge Allina, Vivek Kesar & Joseph A. Odin

  2. Recanati/Miller Transplantation Institute, The Mount Sinai School of Medicine, New York, NY, USA

    Joseph A. Odin

  3. Department of Environmental Medicine, New York University School of Medicine, New York University, Tuxedo, NY, USA

    Jorge Allina & Joseph A. Odin

  4. Department of Pharmacology and Systems Therapeutics, The Mount Sinai School of Medicine, New York, NY, USA

    Jingxiang Bai

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  1. Bin Hu
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  2. Jorge Allina
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  4. Vivek Kesar
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  5. Joseph A. Odin
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Correspondence to Joseph A. Odin.

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Contribution

Bin Hu and Jorge Allina contributed equally to this manuscript. Bin Hu performed the catalase experiments and Jorge Allina performed the estradiol experiments. Vivek Kesar aided with data analysis and manuscript preparation. Jingxiang Bai created the adenoviral vectors and assisted with the catalase experiments. Joseph A. Odin conceptualized and mentored the study and reviewed the final manuscript.

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Hu, B., Allina, J., Bai, J. et al. Catalase and estradiol inhibit mitochondrial protein S-glutathionylation. Mol Cell Biochem 367, 51–58 (2012). https://doi.org/10.1007/s11010-012-1318-7

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  • DOI: https://doi.org/10.1007/s11010-012-1318-7

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