Factor Xa is highly protected from antithrombin-fondaparinux and antithrombin-enoxaparin when incorporated into the prothrombinase complex.
Antithrombin and its cofactor, heparin, target both the product of prothrombin activation by prothrombinase, thrombin, as well as the enzyme responsible for the reaction, factor (F)Xa. These studies were carried out to quantify the effects of each of the prothrombinase components on the half-life of FXa in the presence of antithrombin and the low-molecular-weight heparins (enoxaparin, Aventis, Laval, Quebec, Canada) or the heparin pentasaccharide (fondaparinux, Organon Sanofi-Synthelabo, Cypress, TX, USA). Experiments were carried out using a recombinant form of prothrombin in which the active site serine has been mutated to cysteine and subsequently labeled with fluorescein. This mutant allowed calculation of the second order rate constant for inhibition of FXa by antithrombin in such a way that competition for antithrombin by thrombin is eliminated and competition for FXa by prothrombin is accounted for. Intrinsic rate constants for the inhibition of FXa by antithrombin-enoxaparin and antithrombin-fondaparinux, in the presence of the various prothrombinase components, were calculated. Addition of phospholipid had no significant effect on the second order rate constant for inhibition of FXa by antithrombin, while addition of FVa appeared to be mildly protective. Further addition of prothrombin however, caused profound protection of FXa, increasing its half-life from 1.1 to 353 s in the case of fondaparinux, and from 0.4 to 42 s in the case of enoxaparin. Similar results were reported for unfractionated heparin previously [1]. Therefore, in the presence of unfractionated heparin, fondaparinux, or enoxaparin, prothrombinase is profoundly protected from antithrombin.
Brufatto
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Departments of Biochemistry and Medicine, Queen's University, Kingston, Ontario K7L 3N6, Canada.
Ward
A
A
Nesheim
M E
ME
D000990:Antithrombin III / Q000494:pharmacology*; D001665:Binding Sites / Q000235:genetics; D002384:Catalysis; D004359:Drug Therapy, Combination; D017984:Enoxaparin / Q000494:pharmacology*; D005165:Factor V / Q000187:drug effects*; D015951:Factor Xa / Q000187:drug effects* / Q000378:metabolism; D000077425:Fondaparinux; D006207:Half-Life; D006801:Humans; D007700:Kinetics; D008962:Models, Theoretical; D009154:Mutation; D011134:Polysaccharides / Q000494:pharmacology*; D011516:Prothrombin / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics
D017984:Enoxaparin; D011134:Polysaccharides; D011994:Recombinant Proteins; C022475:prothrombinase complex; D000990:Antithrombin III; D005165:Factor V; D011516:Prothrombin; D015951:Factor Xa; D000077425:Fondaparinux