Chimerism reveals a role for the streptokinase β-domain in nonproteolytic active site formation, substrate, and inhibitor interactions

Inna Gladysheva, Irina Y. Sazonova, Shakeel A. Chowdhry, Lin Liu, Ryan B. Turner, Guy Reed

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Streptokinase (SK) and staphylokinase form cofactor-enzyme complexes that promote the degradation of fibrin thrombi by activating human plasminogen. The unique abilities of streptokinase to nonproteolytically activate plasminogen or to alter the interactions of plasmin with substrates and inhibitors may be the result of high affinity binding mediated by the streptokinase β-domain. To examine this hypothesis, a chimeric streptokinase, SKβswap, was created by swapping the SK β-domain with the homologous β-domain of Streptococcus uberis Pg activator (SUPA or PauA, SK uberis), a streptokinase that cannot activate human plasminogen. SKβswap formed a tight complex with microplasminogen with an affinity comparable with streptokinase. The SKβswap-plasmin complex also activated human plasminogen with catalytic efficiencies (kcat/Km = 16.8 versus 15.2 μM-1 min-1) comparable with streptokinase. However, SKβswap was incapable of nonproteolytic active site generation and activated plasminogen by a staphylokinase mechanism. When compared with streptokinase complexes, SKβswap-plasmin and SKβswap-microplasmin complexes had altered affinities for low molecular weight substrates. The SKβswap-plasmin complex also was less resistant than the streptokinaseplasmin complex to inhibition by α2-antiplasmin and was readily inhibited by soybean trypsin inhibitor. Thus, in addition to mediating high affinity binding to plasmin(ogen), the streptokinase β-domain is required for nonproteolytic active site generation and specifically modulates the interactions of the complex with substrates and inhibitors.

Original languageEnglish (US)
Pages (from-to)26846-26851
Number of pages6
JournalJournal of Biological Chemistry
Volume277
Issue number30
DOIs
StatePublished - Jul 26 2002

Fingerprint

Chimerism
Streptokinase
Catalytic Domain
Substrates
Plasminogen
Fibrinolysin
Antifibrinolytic Agents
Trypsin Inhibitors
Coenzymes
Streptococcus
Fibrin
Soybeans
Thrombosis
Molecular Weight
Molecular weight

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Chimerism reveals a role for the streptokinase β-domain in nonproteolytic active site formation, substrate, and inhibitor interactions. / Gladysheva, Inna; Sazonova, Irina Y.; Chowdhry, Shakeel A.; Liu, Lin; Turner, Ryan B.; Reed, Guy.

In: Journal of Biological Chemistry, Vol. 277, No. 30, 26.07.2002, p. 26846-26851.

Research output: Contribution to journalArticle

Gladysheva, Inna ; Sazonova, Irina Y. ; Chowdhry, Shakeel A. ; Liu, Lin ; Turner, Ryan B. ; Reed, Guy. / Chimerism reveals a role for the streptokinase β-domain in nonproteolytic active site formation, substrate, and inhibitor interactions. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 30. pp. 26846-26851.
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