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Early alpha chain crosslinking in human fibrin preparations
J H Sobel1, C A Thibodeau, R E Canfield
1Department of Medicine, College of Physicians & Surgeons, Columbia University, New York, NY.
Thrombosis and Haemostasis
|October 31, 1988
Summary
Fibrinogen alpha chain crosslinking occurs even with degraded chains, involving regions NH2-terminal to A alpha residue 554. This suggests impaired fibrinogen can still stabilize via crosslinking in pathophysiologic states.
Area of Science:
- Biochemistry
- Molecular Biology
- Hemostasis
Background:
- Fibrinogen is a key protein in blood coagulation.
- Alpha chain crosslinking is crucial for fibrin clot stabilization.
- Degradation of fibrinogen's alpha chains can occur in various diseases.
Purpose of the Study:
- To investigate the role of specific regions in fibrinogen alpha chain crosslinking.
- To determine if partially degraded fibrinogen can still undergo crosslinking.
- To elucidate the early events in alpha chain polymerization.
Main Methods:
- Western blotting (SDS-PAGE) for analyzing alpha polymer evolution.
- Monoclonal antibodies (F-103, F-102) targeting specific A alpha chain regions for immunodetection.
- Immunoaffinity chromatography using F-102 Sepharose to isolate degraded fibrinogen.
Main Results:
- Three crosslinked components (100K, 168K, 210K) showed immunoreactivity with both antibodies.
- Early crosslinking involved interactions between intact and degraded A alpha chains (A alpha #1-276).
- Degraded fibrinogen lacking A alpha #540-554 could form early crosslinked components (95 K, 205 K) and polymers.
Conclusions:
- Early fibrinogen crosslinking activity is localized NH2-terminal to A alpha residue 554.
- Partially degraded fibrinogen molecules can undergo crosslinking and contribute to fibrin stabilization.
- This process occurs even with significant loss (at least 70 residues) of the COOH-terminal A alpha chain.