α Domain deletion converts sreptokinase into a fibrin-dependent plasminogen activator through mechanisms akin to staphylokinase and tissue plasminogen activator

Irina Y. Sazonova, Brian R. Robinson, Inna Gladysheva, Francis J. Castellino, Guy Reed

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The mechanism of action of plasminogen (Pg) activators may affect their therapeutic properties in humans. Streptokinase (SK) is a robust Pg activator in physiologic fluids in the absence of fibrin. Deletion of a "catalytic switch" (SK residues 1-59), alters the conformation of the SK α domain and converts SKΔ59 into a fibrin-dependent Pg activator through unknown mechanisms. We show that the SK α domain binds avidly to the Pg kringle domains that maintain Glu-Pg in a tightly folded conformation. By virtue of deletion of SK residues 1-59, SKΔ59 loses the ability to unfold Glu-Pg during complex formation and becomes incapable of nonproteolytic active site formation. In this manner, SKΔ59 behaves more like staphylokinase than like SK; it requires plasmin to form a functional activator complex, and in this complex SKΔ59 does not protect plasmin from inhibition by α2-antiplasmin. At the same time, SKΔ59 is unlike staphylokinase or SK and is more like tissue Pg activator, because it is a poor activator of the tightly folded form of Glu-Pg in physiologic solutions. SKΔ59 can only activate Glu-Pg when it was unfolded by fibrin interactions or by Cl--deficient buffers. Taken together, these studies indicate that an intact α domain confers on SK the ability to nonproteolytically activate Glu-Pg, to unfold and process Glu-Pg substrate in physiologic solutions, and to alter the substrate-inhibitor interactions of plasmin in the activator complex. The loss of an intact α domain makes SKΔ59 activate Pg through classical "fibrin-dependent mechanisms" (akin to both staphylokinase and tissue Pg activator) that include: 1) a marked preference for a fibrin-bound or unfolded Glu-Pg substrate, 2) a requirement for plasmin in the activator complex, and 3) the creation of an activator complex with plasmin that is readily inhibited by α 2-antiplasmin.

Original languageEnglish (US)
Pages (from-to)24994-25001
Number of pages8
JournalJournal of Biological Chemistry
Volume279
Issue number24
DOIs
StatePublished - Jun 11 2004

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Plasminogen Activators
Plasminogen
Tissue Plasminogen Activator
Streptokinase
Fibrin
Fibrinolysin
Antifibrinolytic Agents
Conformations
Substrates
Kringles
Staphylococcus aureus auR protein
Catalytic Domain
Buffers
Switches
Fluids

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

α Domain deletion converts sreptokinase into a fibrin-dependent plasminogen activator through mechanisms akin to staphylokinase and tissue plasminogen activator. / Sazonova, Irina Y.; Robinson, Brian R.; Gladysheva, Inna; Castellino, Francis J.; Reed, Guy.

In: Journal of Biological Chemistry, Vol. 279, No. 24, 11.06.2004, p. 24994-25001.

Research output: Contribution to journalArticle

Sazonova, Irina Y. ; Robinson, Brian R. ; Gladysheva, Inna ; Castellino, Francis J. ; Reed, Guy. / α Domain deletion converts sreptokinase into a fibrin-dependent plasminogen activator through mechanisms akin to staphylokinase and tissue plasminogen activator. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 24. pp. 24994-25001.
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T1 - α Domain deletion converts sreptokinase into a fibrin-dependent plasminogen activator through mechanisms akin to staphylokinase and tissue plasminogen activator

AU - Sazonova, Irina Y.

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AU - Castellino, Francis J.

AU - Reed, Guy

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AB - The mechanism of action of plasminogen (Pg) activators may affect their therapeutic properties in humans. Streptokinase (SK) is a robust Pg activator in physiologic fluids in the absence of fibrin. Deletion of a "catalytic switch" (SK residues 1-59), alters the conformation of the SK α domain and converts SKΔ59 into a fibrin-dependent Pg activator through unknown mechanisms. We show that the SK α domain binds avidly to the Pg kringle domains that maintain Glu-Pg in a tightly folded conformation. By virtue of deletion of SK residues 1-59, SKΔ59 loses the ability to unfold Glu-Pg during complex formation and becomes incapable of nonproteolytic active site formation. In this manner, SKΔ59 behaves more like staphylokinase than like SK; it requires plasmin to form a functional activator complex, and in this complex SKΔ59 does not protect plasmin from inhibition by α2-antiplasmin. At the same time, SKΔ59 is unlike staphylokinase or SK and is more like tissue Pg activator, because it is a poor activator of the tightly folded form of Glu-Pg in physiologic solutions. SKΔ59 can only activate Glu-Pg when it was unfolded by fibrin interactions or by Cl--deficient buffers. Taken together, these studies indicate that an intact α domain confers on SK the ability to nonproteolytically activate Glu-Pg, to unfold and process Glu-Pg substrate in physiologic solutions, and to alter the substrate-inhibitor interactions of plasmin in the activator complex. The loss of an intact α domain makes SKΔ59 activate Pg through classical "fibrin-dependent mechanisms" (akin to both staphylokinase and tissue Pg activator) that include: 1) a marked preference for a fibrin-bound or unfolded Glu-Pg substrate, 2) a requirement for plasmin in the activator complex, and 3) the creation of an activator complex with plasmin that is readily inhibited by α 2-antiplasmin.

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