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D-dimer specific monoclonal antibodies react with fibrinogen aggregates
A Bennick1, U Haddeland, F Brosstad
1Research Institute for Internal Medicine, Rikshospitalet, Oslo, Norway.
Thrombosis Research
|April 15, 1996
Summary
Heat-induced fibrinogen aggregates, not monomers, mimic fibrin structures. These aggregates enhance tissue plasminogen activator (t-PA) activity, suggesting polymer formation is key for fibrinolysis, even in soluble states.
Area of Science:
- Biochemistry
- Protein Chemistry
- Molecular Biology
Background:
- Human fibrinogen is a key protein in blood coagulation.
- Fibrinogen undergoes conformational changes upon heating.
- Understanding these changes is crucial for fibrinolysis research.
Purpose of the Study:
- To investigate the structural and functional properties of heat-induced fibrinogen aggregates.
- To compare these aggregates with monomeric fibrinogen.
- To elucidate the role of fibrinogen aggregates in fibrinolysis.
Main Methods:
- Gel permeation chromatography to separate fibrinogen fractions.
- Enzyme-linked immunosorbent assay (ELISA) with D-dimer specific antibodies.
- Edman degradation and SDS-PAGE immunoblotting for protein analysis.
Main Results:
- Heat treatment yielded soluble fibrinogen aggregates and monomeric fibrinogen.
- Aggregates, but not monomers, reacted with D-dimer antibodies.
- Both fractions lacked soluble fibrin and high molecular weight derivatives.
- Aggregates, unlike monomers, stimulated t-PA-mediated plasminogen activation.
- Specific peptide sequences (Aalpha-(148-160) and gamma-(312-324)) were exposed in aggregates.
Conclusions:
- Soluble fibrinogen aggregates exhibit fibrin-like structural properties.
- Fibrinogen or fibrin polymer formation is essential for enhanced t-PA activity.
- This effect occurs even when polymers remain in a soluble state, impacting fibrinolysis.