Nitrite ion modifies tyrosine and lysine residues of extracellular matrix proteins

Age-related macular degeneration (AMD) is a disease characterized by degenerative changes in the retinal pigment epithelium and Bruch's membrane. Inflammation is considered a major risk factor for the development and progression of AMD. Nitrite is a potent byproduct of inflammation and has been detected at elevated concentrations in AMD donor tissue. We hypothesize that nitrite chemically modifies the extracellular matrix (ECM) of Bruch's membrane as an initial step to degenerative changes observed in AMD. Non-enzymatically nitrated synthetic ECM peptides, fibronectin and laminin, were used as model systems for inflammation. Using LC/MS, we identified that nitration preferentially occurred on tyrosine and deamination of lysine under the studied conditions. At tyrosine residues, 3-nitrotyrosine was produced and shifted the total mass by the addition of 45 amu. Deamination of lysine occurred and resulted in the formation of either an alkene or alcohol group. The alkene group was observed with a loss of 17 amu. An addition of 1 amu was observed with alcohol formation. We hypothesize that these initial chemical modifications to the structure of ECM proteins may be the responsible for altering the structure and consequent function of Bruch's membrane.