Studies of structural and functional changes of fibrinogen

Czech version

Authors: J. Štikarová; R. Kotlín; J. Suttnar; T. Riedel; J. E. Dyr
Authors‘ workplace: Oddělení biochemie Ústavu hematologie a krevní transfuze Praha, vedoucí oddělení biochemie prof. Ing. Jan E. Dyr, DrSc.
Published in: Vnitř Lék 2012; 58(Suppl 2): 70-83
Category:

Overview

At the Institute of Hematology and Blood Transfusion, we have been studying hereditary dysfibrinogenemia for more than ten years. During this period we have described more than 30 families in the Czech Republic with inherited mutations in fibrinogen. This paper provides an overview of 8 interesting cases of dysfibrinogenemia which we have characterized. Individual cases differ mainly in their clinical manifestations. The study of congenital fibrinogen disorders provides scientists and clinicians important information on structural and functional aspects of the fibrinogen molecule during various physiological processes, especially hemostasis. For roughly one third of the patients with thrombosis the causes of their thrombotic complications have not been found yet. It is therefore possible that at least some of them might be the result of mutations in fibrinogen molecule, especially if these changes do not have to affect basic coagulation tests. Mutations Bβ Arg237Ser, γ Tyr363Asn, and Aα Asn106Asp have thrombotic manifestation. The carriers of these mutations reported both deep vein thrombosis and pulmonary embolism. Mutations Aα Gly13Glu, Aα Arg16Cys, γ Tyr262Cys, and γ Arg275His have bleeding manifestation with varying intensity. A seven year old carrier of the Aα Arg16His mutation has been asymptomatic; however clinical manifestation of the mutation in the future cannot be excluded because the mutation is situated in the site of fibrinopeptide A release. Posttranslational modified fibrinogen is linked with various diseases. These diseases are associated with oxidative stress which leads to uncontrolled production of oxidants. The oxidants modify structure as well as affect function of fibrinogen. We used several oxidative reagents mimicking various (patho)physiological states. We characterized the structural changes with quantification of carbonyls groups and SDS-PAGE followed by immunodetection. Tyrosyl radicals were also detected by SDS-PAGE with immunodetection and by fluorescent determinations. To determine the extent of oxidative nature of the fibrinogen we used OFR (Oxidative Fibrinogen Reactivity). We also studied the influence of these structural changes on the fibrin network architecture (scanning electron microscopy), the interaction of fibrinogen with thrombin (fibrinopeptides release, turbidimetric monitoring of fibrin network formation) and platelets (static and dynamic adhesion of platelets). New carbonyl groups and tyrosyl radicals were formed in fibrinogen. Most modifications occured both to influence fibrin network as well as its interaction with platelets. The overall effect of the functional changes depended on the nature and intensity of the oxidizing agent and what is very important, ranged from protitrombogennic to significantly thrombogenic.

Key words:
fibrinogen – dysfibrinogenemia – oxidative stress

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