Study on the activity of recombinant mutant tissue-type plasminogen activator fused with the C-terminal fragment of hirudin

In the present study, bifunctional fusion proteins were designed by fusing the kringle 2 and protease domains of tissue-type plasminogen activator (tPA) to the C-terminal fragment of hirudin. The thrombolytic and anticoagulant activities of these recombinant proteins from mammalian cells were investigated using in vitro coagulation models and chromogenic assays. The results showed that all assayed tPA mutants retained catalytic activity. The C-terminal fragment of hirudin may have weak affinity to thrombin and thus was insufficient to suppress thrombin-mediated fibrin agglutination. The strength of the thrombolytic activity only relied on the selected tPA sequences, and the fibrinolytic efficiency of single-chain protein significantly decreased. Our data indicate that truncated tPA combined with a hirudin peptide may provide a framework for the further development of a new antithrombotic agent.

Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.

The authors engineered bifunctional fusion proteins linking the kringle 2 and protease domains of tissue-type plasminogen activator to the C-terminal hirudin fragment; all mutants retained catalytic activity, but the hirudin fragment may bind thrombin too weakly to suppress fibrin agglutination.

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