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Related Experiment Videos

Proteolytic alterations of factor Va bound to platelets.

P B Tracy, M E Nesheim, K G Mann

    The Journal of Biological Chemistry
    |January 10, 1983
    PubMed
    Summary

    Platelet-bound Factor Va is cleaved by Factor Xa, generating functional peptides. This cleavage is not essential for Factor Va activity but may enhance its role in thrombin generation.

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    Area of Science:

    • Biochemistry
    • Hematology
    • Molecular Biology

    Background:

    • Factor Va is crucial for thrombin generation, forming the prothrombinase complex on platelet surfaces.
    • Activated protein C (APC) inactivates Factor Va, regulating coagulation.
    • Factor Xa's role in protecting Factor Va from APC and its own proteolytic activity on Factor Va were investigated.

    Purpose of the Study:

    • To investigate the proteolytic events during Factor Va inactivation by APC on platelets.
    • To determine if Factor Xa protects Factor Va from APC.
    • To explore the effect of Factor Xa cleavage on Factor Va function.

    Main Methods:

    • Incubation of Factor Va with washed platelets.
    • Electrophoresis and autoradiography to analyze Factor Va peptides.
    • Proteolytic inactivation studies using APC and Factor Xa.
    • Monitoring thrombin formation and Factor Va cleavage in a phospholipid vesicle model.

    Main Results:

    • Platelet-associated proteases and Factor Xa cleave Factor Va into distinct peptides (D, E, D').
    • APC inactivates platelet-bound Factor Va peptides, yielding five products that remain membrane-associated.
    • Factor Xa protects Factor Va from APC, but active Factor Xa also cleaves Factor Va.
    • Factor Xa rapidly cleaves component E, while APC primarily cleaves components D and D'.
    • Factor Xa-cleaved Factor Va remains catalytically functional.

    Conclusions:

    • Factor Xa cleavage of Factor Va is not essential for its activity in prothrombinase complex.
    • Factor Xa-mediated cleavage may enhance Factor Va's function in thrombin generation.
    • Factor Va undergoes complex proteolytic modifications on the platelet surface by multiple enzymes.

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