Thrombolytic properties of staphylokinase

Matsuo, Osamu; Okada, Katsuyuki; Fukao, Hideharu; Tomioka, Yoshiki; Ueshima, Shigeru; Watanuki, Masaaki; Sakai, Mai

doi:10.1182/blood.v76.5.925.bloodjournal765925

Cited by 15 publications

(20 citation statements)

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“…Clot lysis activity of recombinant proteins in a circulating plasma system in vitro. Thrombolytic activity of r-SAK was compared with that obtained for SAK-RGD-K2-Hirul and SAK-RGD-K2-Hir in the circulating plasma system containing 125 I-labeled thrombus, as described by Matsuo et al (1990). 125 Ilabeled fibrinogen (2 × 10 5 c.p.m.)…”

Section: Methodsmentioning

confidence: 99%

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

Kowalski¹,

Brown²,

Bieniasz³

et al. 2008

Acta Biochim Pol

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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.

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Section: Methodsmentioning

confidence: 99%

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

Kowalski¹,

Brown²,

Bieniasz³

et al. 2008

Acta Biochim Pol

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“…Thrombolytic activity of r‐SAK was compared with that obtained for SAK‐RGD‐K2‐Hir in the circulating plasma system containing [ 125 I]‐labeled thrombus, as described by Matsuo et al [16]. [ 125 I]‐labeled fibrinogen (2 × 10 5 cpm) was used to obtain the radioactive plasma clot.…”

Section: Methodsmentioning

confidence: 99%

IN FOCUS: A new recombinant thrombolytic and antithrombotic agent with higher fibrin affinity – a staphylokinase variant. I. In vitro study

Szemraj

Walkowiak

Kawecka

et al. 2005

Journal of Thrombosis and Haemostasis

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To cite this article: Szemraj J, Walkowiak B, Kawecka I, Janiszewska G, Buczko W, Bartkowiak J, Chabielska E. A new recombinant thrombolytic and antithrombotic agent with higher fibrin affinity -a staphylokinase variant. I. In vitro study. J Thromb Haemost 2005; 3: 2156-65.Summary. We attempted to construct a new recombinant protein characterized by fibrin-specific properties of plasminogen activation combined with antithrombin and antiplatelet activities. To the C-terminal part of recombinant staphylokinase (r-SAK), which is a promising profibrinolytic agent, we assembled: (i) the Kringle 2 domain (K2) of tissue-type plasminogen activator (t-PA), containing a fibrin-specific binding site, (ii) the RGD sequence (Arg-Gly-Asp) for the prevention of platelet aggregation and (iii) the antithrombotic agent -hirudin. The cDNA for hybrid protein SAK-RGD-K2-Hir was cloned into pESP-3 yeast protein expression vector. The introduction of K2 t-PA, RGD sequence and hirudin into r-SAK molecule did not alter the SAK activity.The plasminogen activation rate (determined by K M and K cat ) of SAK-RGD-K2-Hir was not significantly different from that of r-SAK. Affinity and binding strength of the recombinant protein to fibrin immobilized on the biosensor were higher than to r-SAK. We observed a higher clot lysis potency of SAK-RGD-K2-Hir as evidenced by a faster and more profound lysis of 125 I-labeled human fibrin clots. The potency of thrombin inhibition by the hirudin part of the recombinant fusion protein SAK-RGD-K2-Hir was the same as that of r-Hir alone. In conclusion, the results of the in vitro study suggest that the SAK-RGD-K2-Hir construct can be a more potent and faster-acting thrombolytic agent with antithrombin and antiplatelet properties compared with standard r-SAK.Keywords: antiplatelet activity, hirudin and K2 domain of t-PA, recombinant protein, staphylokinase, thrombolysis, thrombolytic and antithrombotic agents.

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“…The concentration of plasma fibrinogen was measured and calculated from standard curves, as described previously. (18) The concentration of plasma fibrinogen decreased to an undetectable level at 1 hour after the first intravenous administration of batroxobin. Higher and larger numbers of doses of batroxobin are lethal in Plg À/À mice.…”

Section: Fibrinogen Depletionmentioning

confidence: 96%

Plasminogen Plays a Crucial Role in Bone Repair

Kawao

Tamura

Okumoto

et al. 2013

Journal of Bone and Mineral Research

105

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The further development in research of bone regeneration is necessary to meet the clinical demand for bone reconstruction. Plasminogen is a critical factor of the tissue fibrinolytic system, which mediates tissue repair in the skin and liver. However, the role of the fibrinolytic system in bone regeneration remains unknown. Herein, we investigated bone repair and ectopic bone formation using plasminogen-deficient (Plg -/-) mice. Bone repair of the femur is delayed in Plg -/-mice, unlike that in the wild-type (Plg þ/þ ) mice. The deposition of cartilage matrix and osteoblast formation were both decreased in Plg -/-mice. Vessel formation, macrophage accumulation, and the levels of vascular endothelial growth factor (VEGF) and transforming growth factor-b (TGF-b) were decreased at the site of bone damage in Plg -/-mice. Conversely, heterotopic ossification was not significantly different between Plg þ/þ and Plg -/-mice. Moreover, angiogenesis, macrophage accumulation, and the levels of VEGF and TGF-b were comparable between Plg þ/þ and Plg -/-mice in heterotopic ossification. Our data provide novel evidence that plasminogen is essential for bone repair. The present study indicates that plasminogen contributes to angiogenesis related to macrophage accumulation, TGF-b, and VEGF, thereby leading to the enhancement of bone repair.

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Thrombolytic properties of staphylokinase

Cited by 15 publications

References 0 publications

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

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

IN FOCUS: A new recombinant thrombolytic and antithrombotic agent with higher fibrin affinity – a staphylokinase variant. I. In vitro study

Plasminogen Plays a Crucial Role in Bone Repair

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