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2009 | 56 | 1 | 41-53
Article title

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

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EN
Abstracts
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.
Publisher

Year
Volume
56
Issue
1
Pages
41-53
Physical description
Dates
published
2009
received
2008-04-24
revised
2008-09-09
accepted
2008-09-18
(unknown)
2008-11-19
Contributors
  • Department of Medical Biochemistry Medical University of Lodz, Łódź, Poland
author
  • Department of Medical Biochemistry Medical University of Lodz, Łódź, Poland
  • Department of Medical Biochemistry Medical University of Lodz, Łódź, Poland
  • Department of Medical Biochemistry Medical University of Lodz, Łódź, Poland
  • Department of Biopharmacy Medical University of Bialystok, Białystok, Poland
  • Department of Biopharmacy Medical University of Bialystok, Białystok, Poland
author
  • Department of Pneumology and Allergy, Medical University of Lodz, Łódź, Poland
  • Department of Medical Biochemistry Medical University of Lodz, Łódź, Poland
  • Department of Medical Biochemistry Medical University of Lodz, Łódź, Poland
  • Department of Medical Biochemistry Medical University of Lodz, Łódź, Poland
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Document Type
Publication order reference
Identifiers
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv56p41kz
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