Small Peptides Blocking Inhibition of Factor Xa and Tissue Factor-Factor VIIa by Tissue Factor Pathway Inhibitor (TFPI)
| Autor: | Markus Fries, Thomas Polakowski, Ulrich Reineke, Michael Dockal, M. Christella L. G. D. Thomassen, Friedrich Scheiflinger, Alexandra C.A. Heinzmann, Hans Brandstetter, Jan Rosing, Rudolf Hartmann, Hartmut J. Ehrlich, Andreas Griessner, Frank Osterkamp |
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| Přispěvatelé: | Biochemie, Ondersteunend personeel CD, RS: CARIM - R1 - Thrombosis and haemostasis |
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Peptide Interactions
Lipoproteins Protein domain Peptide Hemorrhage macromolecular substances Factor VIIa Hemophilia A Biochemistry Protein Structure Secondary Tissue factor Tissue factor pathway inhibitor Protein Domains medicine otorhinolaryngologic diseases Humans Enzyme Inhibitors Mode of action Molecular Biology Reactive center Blood Coagulation chemistry.chemical_classification Hemostasis Coagulants Cell Biology Protease inhibitor (biology) eye diseases Protein Structure Tertiary chemistry Coagulation Factor Xa Biophysics Enzymology Crystal Structure sense organs Peptides Coagulation Factors medicine.drug circulatory and respiratory physiology |
| Zdroj: | Journal of Biological Chemistry, 289(3), 1732-1741. American Society for Biochemistry and Molecular Biology, Inc. |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m113.533836 |
| Popis: | Background: Tissue factor pathway inhibitor (TFPI) inhibits coagulation factors Xa and VIIa. Results: A de novo synthesized 20-mer peptide that binds to TFPI was structurally and functionally characterized. Conclusion: The peptide binds to the Kunitz domain 1 of TFPI and blocks inhibition of factor Xa and factor VIIa by TFPI. Significance: The peptide can potentially prevent bleeding in hemophilia patients. Tissue factor pathway inhibitor (TFPI) is a Kunitz-type protease inhibitor that inhibits activated factor X (FXa) via a slow-tight binding mechanism and tissue factor-activated FVII (TF-FVIIa) via formation of a quaternary FXa-TFPI-TF-FVIIa complex. Inhibition of TFPI enhances coagulation in hemophilia models. Using a library approach, we selected and subsequently optimized peptides that bind TFPI and block its anticoagulant activity. One peptide (termed compound 3), bound with high affinity to the Kunitz-1 (K1) domain of TFPI (K-d approximate to 1 nm). We solved the crystal structure of this peptide in complex with the K1 of TFPI at 2.55- resolution. The structure of compound 3 can be segmented into a N-terminal anchor; an -shaped loop; an intermediate segment; a tight glycine-loop; and a C-terminal -helix that is anchored to K1 at its reactive center loop and two-stranded -sheet. The contact surface has an overall hydrophobic character with some charged hot spots. In a model system, compound 3 blocked FXa inhibition by TFPI (EC50 = 11 nm) and inhibition of TF-FVIIa-catalyzed FX activation by TFPI (EC50 = 2 nm). The peptide prevented transition from the loose to the tight FXa-TFPI complex, but did not affect formation of the loose FXa-TFPI complex. The K1 domain of TFPI binds and inhibits FVIIa and the K2 domain similarly inhibits FXa. Because compound 3 binds to K1, our data show that K1 is not only important for FVIIa inhibition but also for FXa inhibition, i.e. for the transition of the loose to the tight FXa-TFPI complex. This mode of action translates into normalization of coagulation of hemophilia plasmas. Compound 3 thus bears potential to prevent bleeding in hemophilia patients. |
| Databáze: | OpenAIRE |
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