S-Nitrosation of Cellular Proteins by NO Donors in Rat Embryonic Fibroblast 3Y1 Cells: Factors Affecting S-Nitrosation
Figure 2
Protein S-nitrosation induced by exposure to potential S-nitrosating agents in intact 3Y1 cells. (a) Comparison of the efficiencies of potential S-nitrosating agents to S-nitrosate 3Y1 cells. Cells maintained in HBSS were treated with CysNO (200 μM), GSNO (200 μM), NOR-3 (200 μM), or SIN-1 (1 mM) for 30 min. S-nitrosated proteins were detected by the biotin-switch assay. (b and c) Effects of media on S-nitrosation efficiency of S-nitrosothiols. Cells cultured in 10% FCS/DMEM and HBSS were treated with GSNO (1 mM) or CysNO (1 mM) for 30 min, and S-nitrosated proteins were detected by (b) the biotin-switch assay and (c) the DAN assay. The blots shown in (a) and (b) are representative results from several experiments with similar results. Data in (c) are shown as mean ± SEM of three to four independent assays.
