Peroxynitrite induced fibrinogen site identification.

An increasing number of peroxynitrite-mediated fibrinogen nitrifications have been associated with thrombotic diseases. However, few reports related to priority nitrified fibrinogen injury sites exist. In this paper, an improved method, which simulated the environment in vivo, was used to inspect the structural changes of fibrinogen treated with peroxynitrite and LC-MS/MS in order to investigate the fibrinogen injury sites. The SDS-PAGE results indicated that γ subunits of Fg were vulnerable to oxidative/nitrative modifications induced by peroxynitrite. An in-depth analysis of fibrinogen γ chain (Fg I) nitration site identification and susceptibility to peroxynitrite-utilizing LC-MS/MS strategy was performed. Based on a large dataset, the results indicated the priority injury sites during database searching were Y96, Y262, Y274, Y348, and Y363; these results could be applied to biomedical studies.