Figure 1.

The ANSID approach for aromatic nitration site identification. A schematic of the ANSID method. Proteins are first digested with trypsin, followed by Lys- and N-terminal amine-blocking through reductive dimethylation with formaldehyde and dimethylamine borane. The nitro-aromatic residues are then reduced to produce amines with hemin-agarose beads in the presence of DTT, followed by capture and enrichment of the amino-aromatic amino acid-containing peptides using aldehyde-agarose beads. The captured peptides are then eluted with TFA and analyzed by nanoLC-MS/MS. Abbreviations: DMAB, dimethylamine borane; DTT, dithiothreitol; TFA, trifluoroacetic acid.