Effect of Phenylglyoxal-modified α₂-Antiplasmin on Urokinase-induced Fibrinolysis

Summary

One of the functions of activated blood clotting factor XIII (FXIIIa) is the crosslinking of α₂-antiplasmin (α₂AP) to fibrin. This process results in localization and concentration of α₂AP throughout fibrin, thereby making fibrin more resistant to digestion by plasmin. We reasoned that competition by chemically-modified inactive α₂AP (modα₂AP) with native α₂AP would diminish the resistance of fibrin to digestion by plasmin. Modα₂AP was prepared by treating native α₂AP with an Arg-specific reagent, phenylglyoxal. An average of four of the total nineteen Arg residues in α₂AP reacted with phenylglyoxal and resulted in complete loss of plasmin inhibitory activity; however, modα₂AP competed effectively with native α₂AP for becoming crosslinked to fibrin by FXIIIa catalysis. In the presence of modα₂AP, urokinase (UK)-induced plasma clot lysis time shortened significantly. Modα₂AP enhanced UK-induced clot lysis in a whole blood system as shown by the similarities of rates of clot lysis for a mixture of 20 U/ml UK and 1.5 μM modα₂AP versus that induced by 100 U/ml UK without modα₂AP. Less fibrinogenolysis occurred in whole blood when modα₂AP was present since much lower UK concentrations were needed to achieve the same level of fibrinolysis than when only native α₂AP was present. Our results indicate that modα₂AP enhances UK-induced fibrinolysis by competitive inhibition of factor XIIIa-mediated incorporation of native α₂AP into fibrin.

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