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Effects of high NaCl on cellular glutathione and glutathionylation of proteins in HeLa cells
Author(s) -
Edwards Sarah K.,
Burg Maurice B.,
Dmitrieva Natalia I.
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.859.2
Subject(s) - glutathione , hela , reactive oxygen species , oxidative stress , chemistry , apoptosis , antioxidant , microbiology and biotechnology , blot , protein carbonylation , biochemistry , biology , cell , gene , enzyme
High NaCl causes oxidative stress, evidenced by increased cellular reactive oxygen species (ROS) and oxidation (carbonylation) of proteins. Reduced glutathione neutralizes ROS by becoming oxidized. Reversible glutathionylation of cysteines in proteins protects the proteins from irreversible oxidation and signals the redox state. We measured glutathione in HeLa cells and analyzed glutathionylation of proteins with anti‐glutathione antibody in non‐reducing Western blots. HeLa cells survive modest increases of NaCl (to 400–500 mosmol/kg), but die from apoptosis at 600–700 mosmol/kg. Acute increase to 400–500 mosmol/kg for 2 hours has little effect on the level of glutathione, but increase to 600–700 mosmol/kg decreases glutathione level at 1 hour, before the cells become apoptotic, and completely eliminates it by 2 hours, coincident with activation of apoptosis. Elevation of NaCl to 400–500 mosmol/kg, but not to 600–700 mosmol/kg, increases glutathionylation of a 40–45 kDa protein. Imminoprecipitation with anti‐glutathione antibody reveals several additional proteins whose glutathionylation increases at 400 mosmol/kg. We conclude that high NaCl‐induced apoptosis in HeLa cells is associated with a decreased level of glutathione, and we speculate that protein glutathionylation might contribute to signaling cellular adaptation to high NaCl. Supported by the Intramural Program of NHLBI.