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

Abstract:

: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 (125)I-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.

journal_name

Acta Biochim Pol

journal_title

Acta biochimica Polonica

authors

Kowalski M,Brown G,Bieniasz M,Oszajca K,Chabielska E,Pietras T,Szemraj Z,Makandjou-Ola E,Bartkowiak J,Szemraj J

subject

Has Abstract

pub_date

2009-01-01 00:00:00

pages

41-53

issue

1

eissn

0001-527X

issn

1734-154X

pii

20081653

journal_volume

56

pub_type

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