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Prepublished online as a Blood First Edition Paper on March 27, 2003; DOI 10.1182/blood-2002-10-3150.

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2002-10-3150v1
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Blood, 15 July 2003, Vol. 102, No. 2, pp. 535-540

HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY

Fibrinogen gamma-chain splice variant {gamma}' alters fibrin formation and structure

Amy V. Cooper, Kristina F. Standeven, and Robert A. S. Ariëns

From the Academic Unit of Molecular Vascular Medicine, University of Leeds, Martin Wing, Leeds General Infirmary, United Kingdom.

Fibrinogen {gamma}A/{gamma}' results from alternative splicing of mRNA. This variant, which constitutes approximately 8% to 15% of plasma fibrinogen, contains FXIII and thrombin binding sites. Our objective was to investigate whether {gamma}A/{gamma}' differs in fibrin formation and structure from the more common variant {gamma}A/{gamma}A. Both variants were separated and purified by anion-exchange chromatography. Fibrin formation and clot structure of the variants and unfractionated fibrinogen were investigated by turbidity and scanning electron microscopy (SEM). Thrombin cleavage of fibrinopeptides was analyzed by high-performance liquid chromatography (HPLC). Turbidity analysis showed significantly altered polymerization rates and overall fiber thickness in {gamma}A/{gamma}' clots compared with {gamma}A/{gamma}A and unfractionated fibrinogen. This finding was consistent with a range of thrombin concentrations. HPLC demonstrated reduced rates of fibrinopeptide B (FpB) release from {gamma}A/{gamma}' fibrinogen compared with {gamma}A/{gamma}A. Delayed FpB release was associated with delayed lateral aggregation of protofibrils and significant differences were found on SEM, with {gamma}A/{gamma}' clots consisting of smaller diameter fibers and increased numbers of branch points compared with both {gamma}A/{gamma}A and unfractionated fibrinogen. These results demonstrate that the {gamma}A/{gamma}' splice variant of fibrinogen directly alters fibrin formation and structure, which may help to explain the increased thrombotic risk associated with this variant.


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