Von Willebrand factor-binding protein is a hysteretic conformational activator of prothrombin.
Von Willebrand factor-binding protein (VWbp), secreted by Staphylococcus aureus, displays secondary structural homology to the 3-helix bundle, D1 and D2 domains of staphylocoagulase (SC), a potent conformational activator of the blood coagulation zymogen, prothrombin (ProT). In contrast to the classical proteolytic activation mechanism of trypsinogen-like serine proteinase zymogens, insertion of the first 2 residues of SC into the NH(2)-terminal binding cleft on ProT (molecular sexuality) induces rapid conformational activation of the catalytic site. Based on plasma-clotting assays, the target zymogen for VWbp may be ProT, but this has not been verified, and the mechanism of ProT activation is unknown. We demonstrate that VWbp activates ProT conformationally in a mechanism requiring its Val(1)-Val(2) residues. By contrast to SC, full time-course kinetic studies of ProT activation by VWbp demonstrate that it activates ProT by a substrate-dependent, hysteretic kinetic mechanism. VWbp binds weakly to ProT (K(D) 2.5 microM) to form an inactive complex, which is activated through a slow conformational change by tripeptide chromogenic substrates and its specific physiological substrate, identified here as fibrinogen (Fbg). This mechanism increases the specificity of ProT activation by delaying it in a slow reversible process, with full activation requiring binding of Fbg through an exosite expressed on the activated ProT*.VWbp complex. The results suggest that this unique mechanism regulates pathological fibrin (Fbn) deposition to VWF-rich areas during S. aureus endocarditis.
Kroh
Heather K
HK
Department of Pathology, Vanderbilt University School of Medicine, Nashville, TN 37232-2561, USA.
Panizzi
Peter
P
Bock
Paul E
PE
D001667:Binding, Competitive; D001777:Blood Coagulation; D002352:Carrier Proteins / Q000737:chemistry / Q000502:physiology*; D004789:Enzyme Activation; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008968:Molecular Conformation; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011516:Prothrombin / Q000737:chemistry*; D013211:Staphylococcus aureus / Q000378:metabolism; D013379:Substrate Specificity; D014362:Trypsinogen / Q000737:chemistry; D014841:von Willebrand Factor / Q000737:chemistry / Q000502:physiology*
D002352:Carrier Proteins; D014841:von Willebrand Factor; D011516:Prothrombin; D014362:Trypsinogen