TOXI 80 |
| Tyrosine nitration is an important post-translational modification that is associated with oxidative stress and activation of nitric oxide synthases, leading to increased production of nitric oxide. Described here is a method that combines the specific isolation and enrichment of nitrated peptides through immunoprecipitation with mass spectrometric identification of the peptide sequences. Using nitrated human serum albumin as a model system, the major nitrated peptides were enriched and positively identified by mass spectrometric methods including confirmation of structure by peptide mass fingerprinting and LC/MS/MS, with data analysis by MASCOT and Spectrum Mill. Additionally, this method was applied to the analysis of hepatocytes that had been activated to produce nitric oxide which, through formation of other reactive nitrogen species, leads to tyrosine nitration. The inducible nitric oxide synthase (iNOS) was stimulated by a mixture containing an endotoxin (lipopolysaccharide), cytokines (interferon , interleukin-1, and tumor necrosis factor. Identified nitrated proteins will be presented. |
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Poster Presentations and Awards
6:00 PM-10:00 PM, Tuesday, 12 September 2006 Moscone Center -- Room 104, Poster
Sci-Mix
Division of Chemical Toxicology |