Inhibition of factor X and factor V activation by dermatan sulfate and a pentasaccharide with high affinity for antithrombin III in human plasma.
There is evidence that by catalyzing thrombin inhibition, several glycosaminoglycans can inhibit the thrombin-mediated amplification reactions of coagulation and thereby delay prothrombin activation. The two amplification reactions can apparently be catalysed by endogenously generated factor Xa and thrombin. This study provides evidence which suggests that on a molar basis, an agent which can only catalyse thrombin inhibition is approximately 10 times more effective than an agent which can only catalyse factor Xa inhibition in their ability to inhibit intrinsic prothrombin activation. We determined the concentrations of each of heparin, dermatan sulfate and a pentasaccharide with high affinity for antithrombin III, to delay intrinsic prothrombin activation for at least 15s. Heparin catalyses both thrombin and factor Xa inhibition; dermatan sulfate catalyses only thrombin inhibition, while the pentasaccharide only catalyses factor Xa inhibition. Efficient prothrombin activation, which coincided with both factor X activation and factor V proteolysis, was first observed 45s after CaC12 was added to contact-activated plasma. Heparin (approximately 0.1 microM) prolonged by at least 30 s the time required for the activation of the three clotting factors to begin. The minimum concentrations of the pentasaccharide and dermatan sulfate to delay the activation of prothrombin, factors X and V were approximately 50 microM and approximately 5 microM, respectively. Thus, each anticoagulant could inhibit intrinsic prothrombin activation only when it inhibited activation of both factors X and V. A combination of approximately 5 microM pentasaccharide and approximately 0.05 microM dermatan sulfate similarly delayed the activation of all three clotting factors. Thus, while catalysis of thrombin inhibition is a more effective pathway than catalysis of factor Xa inhibition for delaying prothrombin activation, the simultaneous catalysis of thrombin and factor Xa inhibition can synergistically improve the ability of a sulfated polysaccharide to delay prothrombin activation.
Ofosu
F A
FA
Canadian Red Cross Society, Blood Transfusion Service, Hamilton.
Choay
J
J
Anvari
N
N
Smith
L M
LM
Blajchman
M A
MA
D000818:Animals; D000990:Antithrombin III / Q000378:metabolism*; D002122:Calcium Chloride / Q000494:pharmacology; D002240:Carbohydrate Sequence; D002384:Catalysis; D003871:Dermatan Sulfate / Q000494:pharmacology*; D004357:Drug Synergism; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases; D004797:Enzyme-Linked Immunosorbent Assay; D005165:Factor V / Q000037:antagonists & inhibitors*; D005170:Factor X / Q000037:antagonists & inhibitors*; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D011516:Prothrombin / Q000378:metabolism; D012756:Sheep; D013917:Thrombin / Q000378:metabolism
D011134:Polysaccharides; D003871:Dermatan Sulfate; D000990:Antithrombin III; D005165:Factor V; D011516:Prothrombin; D005170:Factor X; D006493:Heparin; D010450:Endopeptidases; D013917:Thrombin; D002122:Calcium Chloride