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Prothrombin conversion under flow conditions by prothrombinase assembled on adherent platelets

D Billy1, J Briedé, J W Heemskerk

  • 1Department of Biochemistry, Maastricht University, The Netherlands.

Blood Coagulation & Fibrinolysis : an International Journal in Haemostasis and Thrombosis
|April 1, 1997
PubMed
Summary

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Platelet activation on fibrinogen surfaces supports stable prothrombinase complex assembly. Thrombin generation is primarily limited by prothrombin supply, not factor Va concentration.

Area of Science:

  • Biochemistry
  • Hematology
  • Cell Biology

Background:

  • Platelets play a crucial role in hemostasis and thrombosis.
  • Prothrombinase complex (factors Xa and Va) is essential for thrombin generation.
  • Understanding prothrombinase assembly on activated platelets is key to hemostasis research.

Purpose of the Study:

  • To investigate prothrombin activation by prothrombinase on adhered platelets.
  • To determine the factors limiting thrombin formation on activated platelets in flow conditions.

Main Methods:

  • Utilized a flow chamber model with fibrinogen-coated coverslips.
  • Observed platelet morphology changes and annexin V binding to detect procoagulant phospholipids.
  • Assembled prothrombinase complexes on adhered platelets by perfusing with factors Xa, Va, and prothrombin.

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Main Results:

  • Adherent platelets undergo morphological changes, including pseudopod formation and spreading, culminating in balloon-shaped cells.
  • Calcium-dependent exposure of procoagulant phospholipids on the outer membrane was observed.
  • Thrombin formation rate increased with prothrombin concentration but plateaued with factor Va (0.05-0.5 nM).
  • Factor Xa could be omitted post-assembly without affecting steady-state thrombin production.

Conclusions:

  • Adherent platelets provide an efficient procoagulant surface for stable prothrombinase complex assembly.
  • Thrombin production is primarily limited by the rate of prothrombin supply to the catalytic surface.
  • This study elucidates the dynamics of thrombin generation on activated platelets under flow.