The effect of chemical anti-inhibitors on fibrinolytic enzymes and inhibitors.
Fibrinolytic enzyme inhibitors hamper the determination of the specific fibrinolytic serine protease activity. Reportedly, chemical anti-inhibitors eliminate the influence of fibrinolytic inhibitors, but it remains unclear to what extent they change the specific activity of fibrinolytic serine proteases. We studied the influence of chemical anti-inhibitors (chloramine T, flufenamate, sodium lauryl sulfate, and methylamine) on fibrinolytic serine proteases and fibrinolytic enzyme inhibitors using the physiological substrate fibrin as plasmin substrate. Low concentrations of chloramine T (0.01 mmol/l) prevent the inhibition of plasminogen activators. Higher concentrations (1 mmol/l) reduce the inhibition of plasmin, but simultaneously quench the plasminogen activator activity. Flufenamate eliminates most fibrinolytic enzyme inhibitors, but increases the activity of plasmin (apparent recovery 140%) and plasminogen activators (apparent recovery > 200%). Sodium lauryl sulfate eliminates the major fibrinolytic enzyme inhibitors, but increases the activity of plasmin (apparent recovery > 200%) and plasminogen activator, urokinase type (apparent recovery 130%). Methylamine affects only plasmin inhibition. We conclude that chemical anti-inhibitors may invalidate the analytical specificity of methods for the determination of fibrinolytic serine protease activity.
Sidelmann
J
J
Institute for Thrombosis Research, South Jutland University Centre, Ribe County Hospital, Esbjerg, Denmark. thromsuc@inet.uni-c.dk
Jespersen
J
J
Kluft
C
C
Gram
J
J
D002700:Chloramines / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005341:Fibrinolysin / Q000037:antagonists & inhibitors / Q000737:chemistry; D005342:Fibrinolysis / Q000187:drug effects*; D005343:Fibrinolytic Agents / Q000097:blood / Q000494:pharmacology*; D005439:Flufenamic Acid / Q000494:pharmacology; D006801:Humans; D008744:Methylamines / Q000494:pharmacology; D017395:Plasminogen Activator Inhibitor 1 / Q000097:blood / Q000494:pharmacology; D010960:Plasminogen Activators / Q000037:antagonists & inhibitors / Q000737:chemistry; D015842:Serine Proteinase Inhibitors / Q000097:blood*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D010959:Tissue Plasminogen Activator / Q000037:antagonists & inhibitors / Q000097:blood / Q000187:drug effects; D014105:Tosyl Compounds / Q000494:pharmacology; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors / Q000097:blood / Q000187:drug effects
D002700:Chloramines; D005343:Fibrinolytic Agents; D008744:Methylamines; D017395:Plasminogen Activator Inhibitor 1; D015842:Serine Proteinase Inhibitors; D014105:Tosyl Compounds; C016300:chloramine-T; D012967:Sodium Dodecyl Sulfate; D005439:Flufenamic Acid; C027451:methylamine; D010960:Plasminogen Activators; D010959:Tissue Plasminogen Activator; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator