title: Activity Regulation by Fibrinogen and Fibrin of Streptokinase from Streptococcus Pyogenes.
abstract: Streptokinase is a virulence factor of streptococci and acts as a plasminogen activator to generate the serine protease plasmin which promotes bacterial metastasis. Streptokinase isolated from group C streptococci has been used therapeutically as a thrombolytic agent for many years and its mechanism of action has been extensively studied. However, group A streptococci are associated with invasive and potentially fatal infections, but less detail is available on the mechanism of action of streptokinase from these bacteria. We have expressed recombinant streptokinase from a group C strain to investigate the therapeutic molecule (here termed rSK-H46A) and a molecule isolated from a cluster 2a strain from group A (rSK-M1GAS) which is known to produce the fibrinogen binding, M1 protein, and is associated with life-threatening disease. Detailed enzyme kinetic models have been prepared which show how fibrinogen-streptokinase-plasminogen complexes regulate plasmin generation, and also the effect of fibrin interactions. As is the case with rSK-H46A our data with rSK-M1GAS support a "trigger and bullet" mechanism requiring the initial formation of SK•plasminogen complexes which are replaced by more active SK•plasmin as plasmin becomes available. This model includes the important fibrinogen interactions that stimulate plasmin generation. In a fibrin matrix rSK-M1GAS has a 24 fold higher specific activity than the fibrin-specific thrombolytic agent, tissue plasminogen activator, and 15 fold higher specific activity than rSK-H46A. However, in vivo fibrin specificity would be undermined by fibrinogen stimulation. Given the observed importance of M1 surface receptors or released M1 protein to virulence of cluster 2a strain streptococci, studies on streptokinase activity regulation by fibrin and fibrinogen may provide additional routes to addressing bacterial invasion and infectious diseases.
authors.affiliation: Component development laboratory, NHS Blood and Transplant, Cambridge Donor Centre, Cambridge, United Kingdom.
authors.forename: Sian
authors.initials: S
authors.lastname: Huish
authors.affiliation: Biotherapeutics Section, National Institute for Biological Standard and Control, South Mimms, Herts, United Kingdom.
authors.forename: Craig
authors.initials: C
authors.lastname: Thelwell
authors.affiliation: Biotherapeutics Section, National Institute for Biological Standard and Control, South Mimms, Herts, United Kingdom.
authors.forename: Colin
authors.identifier: http://orcid.org/0000-0001-7608-208X
authors.initials: C
authors.lastname: Longstaff
mesh_terms: D005337:Fibrin / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D008962:Models, Theoretical; D010958:Plasminogen / Q000378:metabolism; D013297:Streptococcus pyogenes / Q000378:metabolism*; D013300:Streptokinase / Q000378:metabolism*
chemical_list: D005337:Fibrin; D005340:Fibrinogen; D010958:Plasminogen; D013300:Streptokinase