The susceptibility of plasma coagulation factor XI to nitration and peroxynitrite action.
Coagulation factor XI is present in blood plasma as the zymogen, like other serine proteases of hemostatic system, but as the only coagulation factor forms 140-160kDa homodimers. Its activation is induced by thrombin, and a positive feedback increases the generation of the extra thrombin. Experimental and clinical observations confirm protective roles of factor XI deficiencies in certain types of thromboembolic disorders. Thromboembolism still causes serious problems for modern civilization. Diseases associated with the blood coagulation system are often associated with inflammation and oxidative stress. Peroxynitrite is produced from nitric oxide and superoxide in inflammatory diseases. The aim of the current study is to evaluate effects of nitrative stress triggered by peroxynitrite on coagulation factor XI in human plasma employing biochemical and bioinformatic methods. The amidolytic assay shows increase in factor XI activity triggered by peroxynitrite. Peroxynitrite interferes factor XI by nitration and fragmentation, which is demonstrated by immunoprecipitation followed by western blotting. Nitrated factor XI is even present in control blood plasma. The results suggest possible modifications of factor XI on the molecular level. Computer simulations show tyrosine residues as targets of peroxynitrite action. The modifications induced by peroxynitrite in factor XI might be important in thrombotic disorders.
Department of General Biochemistry, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland. Electronic address: mponczek@biol.uni.lodz.pl.
Michał Błażej
MB
Ponczek
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D017124:Conserved Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D019143:Evolution, Molecular; D005172:Factor XI / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D008024:Ligands; D062105:Molecular Docking Simulation; D015538:Nitrosation; D010314:Partial Thromboplastin Time; D030421:Peroxynitrous Acid / Q000494:pharmacology*; D010949:Plasma / Q000378:metabolism; D016415:Sequence Alignment; D013379:Substrate Specificity / Q000187:drug effects; D013917:Thrombin / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
D005456:Fluorescent Dyes; D008024:Ligands; D030421:Peroxynitrous Acid; D014443:Tyrosine; D005172:Factor XI; D013917:Thrombin
3-nitrotyrosine; Coagulation factor XI; Peroxynitrite