Peroxynitrite-mediated modification of fibrinogen affects platelet aggregation and adhesion.
The reaction of peroxynitrite with fibrinogen resulted in both structural modifications and altered biological properties of this glycoprotein. SDS-PAGE analysis of peroxynitrite-treated fibrinogen, performed under non-reducing conditions, showed some aggregated material on the top of the gel (5-10% of total staining bands) and the presence of nitrotyrosine. The amount of nitrotyrosine, detected by immunoassay with anti-nitrotyrosine antibodies, was dependent on peroxynitrite concentration. In comparison with native molecule, peroxynitrite-treated fibrinogen subjected to SDS-PAGE under reducing conditions revealed not only three bands corresponding to Aalpha, Bbeta and gamma chains, but the existence of additional high molecular bands probably due to the formation of dityrosine crosslinking between fibrinogen subunits. The different susceptibility in tyrosine nitration of fibrinogen subunits was also observed. The Aalpha chain was the most intensely nitrated, while Bbeta and gamma chains were nitrated much less? Peroxynitrite-treated fibrinogen in comparison with native molecule had a distinct capability to mediate platelet adhesion and aggregation. Both unstimulated and ADP-activated platelets showed a reduced ability to adhere to peroxynitrite-modified fibrinogen. The percentage of ADP-induced platelet aggregation decreased as a function of peroxynitrite-mediated modification of fibrinogen molecule.
Nowak
Pawel
P
Department of General Biochemistry, University of Lodz, Lodz, Poland. pnowak@biol.lodz.pl
Wachowicz
Barbara
B
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000818:Animals; D004305:Dose-Response Relationship, Drug; D005340:Fibrinogen / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D016877:Oxidants / Q000494:pharmacology*; D030421:Peroxynitrous Acid / Q000494:pharmacology*; D010973:Platelet Adhesiveness / Q000187:drug effects*; D010974:Platelet Aggregation / Q000187:drug effects*; D010979:Platelet Function Tests; D013552:Swine
D016877:Oxidants; D030421:Peroxynitrous Acid; D000244:Adenosine Diphosphate; D005340:Fibrinogen