Selective modulation of thrombin‐activatable fibrinolysis inhibitor (TAFI) activation by thrombin or the thrombin‐thrombomodulin complex using TAFI‐derived peptides. (30th September 2015)

• Record Type: Journal Article
• Authors: Plug, T.; Marquart, J. A.; Marx, P. F.; Meijers, J. C. M.
• Is Part Of: Journal of thrombosis and haemostasis. Volume 13:Number 11(2015:Nov.)
• Page Start: 2093
• Page End: 2101
• Publication Date: 2015-09-30
• Subjects: carboxypeptidase B2 -- fibrinolysis -- thrombin -- thrombin‐activatable fibrinolysis inhibitor -- thrombomodulin; Thrombosis -- Periodicals; Hemostasis -- Periodicals; Blood coagulation disorders -- Periodicals; 616.1
• DOI: 10.1111/jth.13133
• Languages: English
• ISSNs: 1538-7933

Abstract

Summary: Background: Thrombin‐activatable fibrinolysis inhibitor (TAFI) is a risk factor for coronary heart disease. TAFI is proteolytically activated by thrombin, the thrombin‐thrombomodulin complex and plasmin. Once active, it dampens fibrinolysis and inflammation. The aim of this study was to generate TAFI‐derived peptides that specifically modulate TAFI activation and activity. Methods: Thirty‐four overlapping TAFI peptides, and modifications thereof, were synthesized. The effects of these peptides on TAFI activation and TAFIa activity were determined. In addition, the binding of the peptides to thrombin were determined. Results: Four peptides (peptides 2, 18, 19 and 34) inhibited TAFI activation and two peptides (peptides 14 and 24) inhibited TAFIa activity directly. Peptide 2 (Arg12‐Glu28) and peptide 34 (Cys383‐Val401) inhibited TAFI activation by the thrombin‐thrombomodulin complex with IC50 values of 7.3 ± 1.8 and 6.1 ± 0.9 μm, respectively. However, no inhibition was observed in the absence of thrombomodulin. This suggests that the regions Arg12‐Glu28 and Cys383‐Val401 in TAFI are involved in thrombomodulin‐mediated TAFI activation. Peptide 18 (Gly205‐Ser221) and peptide 19 (Arg214‐Asp232) inhibited TAFI activation by thrombin and the thrombin‐thrombomodulin complex. Furthermore, these peptides bound to thrombin ( K D : 1.5 ± 0.4 and 0.52 ± 0.07 μm for peptides 18 and 19, respectively), suggesting that Gly205‐Asp232 of TAFI is involved in binding to thrombin. Peptide 14 (His159‐His175) inhibited TAFIa activity. The inhibition was TAFIa specific, because no effect on the homologous enzyme carboxypeptidase B was observed. Conclusions: Thrombin‐activatable fibrinolysis inhibitor‐derived peptides show promise as new tools to modulate TAFI activation and TAFIa activity. Furthermore, these peptides revealed potential binding sites on TAFI for thrombin and the thrombin‐thrombomodulin complex.