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Redox Biol. 2014 Jan 15;2:436-46. doi: 10.1016/j.redox.2013.12.030. eCollection 2014.

Identification of DUOX1-dependent redox signaling through protein S-glutathionylation in airway epithelial cells.

Author information

1
Department of Pathology, Vermont Lung Center, College of Medicine, University of Vermont, Burlington, VT 05405, United States.
2
Department of Biology, College of Arts and Sciences, University of Vermont, Burlington, VT 05405, United States.
3
Department of Physiology, Faculty of Medicine, and "Lendulet" Peroxidase Enzyme Research Group, Semmelweis University, Budapest, Hungary.

Abstract

The NADPH oxidase homolog dual oxidase 1 (DUOX1) plays an important role in innate airway epithelial responses to infection or injury, but the precise molecular mechanisms are incompletely understood and the cellular redox-sensitive targets for DUOX1-derived H2O2 have not been identified. The aim of the present study was to survey the involvement of DUOX1 in cellular redox signaling by protein S-glutathionylation, a major mode of reversible redox signaling. Using human airway epithelial H292 cells and stable transfection with DUOX1-targeted shRNA as well as primary tracheal epithelial cells from either wild-type or DUOX1-deficient mice, DUOX1 was found to be critical in ATP-stimulated transient production of H2O2 and increased protein S-glutathionylation. Using cell pre-labeling with biotin-tagged GSH and analysis of avidin-purified proteins by global proteomics, 61 S-glutathionylated proteins were identified in ATP-stimulated cells compared to 19 in untreated cells. Based on a previously established role of DUOX1 in cell migration, various redox-sensitive proteins with established roles in cytoskeletal dynamics and/or cell migration were evaluated for S-glutathionylation, indicating a critical role for DUOX1 in ATP-stimulated S-glutathionylation of β-actin, peroxiredoxin 1, the non-receptor tyrosine kinase Src, and MAPK phosphatase 1. Overall, our studies demonstrate the importance of DUOX1 in epithelial redox signaling through reversible S-glutathionylation of a range of proteins, including proteins involved in cytoskeletal regulation and MAPK signaling pathways involved in cell migration.

KEYWORDS:

Cell migration; Cysteine; DUOX1; NADPH oxidase; Proteomics; S-glutathionylation

PMID:
24624333
PMCID:
PMC3949091
DOI:
10.1016/j.redox.2013.12.030
[Indexed for MEDLINE]
Free PMC Article

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