Cloning and expression of a new recombinant thrombolytic and antithrombotic agent - a staphylokinase variant

• Autorzy: Kowalski M. , Brown G. , Bieniasz M. , Oszajca K. , Chabielska E. , Pietras T. , Szemraj Z. , Makandjou-Ola E. , Bartkowiak J. , Szemraj J.
• Języki publikacji: EN

Abstrakty

EN

 To develop a more potent antithrombin agent with thrombolytic and antiplatelet properties, a new staphylokinase (SAK) variant was constructed. The kringle 2 domain (K2) of tissue type-plasminogen activator (t-PA) containing a fibrin-specific binding site (i), the RGD sequence (Arg-Gly-Asp) for the prevention of platelet aggregation (ii) and the antithrombotic agent - hirulog (iii) was assembled to the C-terminal part of recombinant staphylokinase (r-SAK). cDNA for the hybrid protein SAK-RGD-K2-Hirul was cloned into Pichia pastoris pPIC9K yeast expression vector. The introduction of K2 t-PA, the RGD sequence and hirulog into the C-terminus of r-SAK did not alter the staphylokinase activity. We observed a higher clot lysis potency of SAK-RGD-K2-Hirul as evidenced by a faster and more profound lysis of 125I-labeled human fibrin clots. The potency of thrombin inhibition by the hirulog C-terminal part of the recombinant fusion protein was almost identical to that of r-Hir alone. These results suggest that the SAK-RGD-K2-Hirul construct can be a more potent and faster-acting thrombolytic agent with better antithrombin and antiplatelet properties compared to r-SAK and SAK-RGD-K2-Hir.

Słowa kluczowe

EN

antithrombotic agent zob.fibrinolytic agent; thrombolytic agent zob.fibrinolytic agent; fibrinolytic agent; thrombolysis; recombinant protein; expression; recombinant fusion protein; antiplatelet activity; staphylokinase; cloning; new agent; coronary heart disease; platelet aggregation; thromboembolism

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2009
• Tom: 56
• Numer: 1
• Opis fizyczny: p.41-53,fig.,ref.

Twórcy

autor

Kowalski M.

• Medical University of Lodz, Mazowiecka 6/8, 92-215 Lodz, Poland

autor

Brown G.

autor

Bieniasz M.

autor

Oszajca K.

autor

Chabielska E.

autor

Pietras T.

autor

Szemraj Z.

autor

Makandjou-Ola E.

autor

Bartkowiak J.

autor

Szemraj J.

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