Investigation of Thrombin Activity with PAR 1-based Fluorogenic Peptides
| Autor: | Julio A. Mignaco, Maria Aparecida Julianod, Saulo Martins Vieira, Flavia Garcia dos Reis, Russolina B. Zingali, Reinaldo B Geraldo, Luiz Juliano, Denis L. S. Dutra |
|---|---|
| Předmět: |
medicine.medical_treatment
Molecular Sequence Data Peptide Biochemistry Substrate Specificity Thrombin Structural Biology medicine Fluorescence Resonance Energy Transfer Humans Receptor PAR-1 Amino Acid Sequence Enzyme Assays Fluorescent Dyes chemistry.chemical_classification Protease Hydrolysis Substrate (chemistry) General Medicine Amino acid Molecular Docking Simulation Förster resonance energy transfer Coagulation chemistry Ionic strength Peptides medicine.drug |
| Zdroj: | ResearcherID |
| Popis: | Thrombin, a highly specific protease of blood coagulation, has two exosites that modulate its specificity. We designed two sets of synthetic substrate FRET peptides with 25- or 11- amino acids (aa) each, based on the PAR 1 sequence, to characterize the effect of exosite 1 engagement on substrate catalysis and preference. The 25-aa set encompassed a sequence binding to exosite 1, and structural modeling showed that binding to thrombin did not differ significantly from that of PAR 1 peptide. Modification at the P3´position of the 25 or 11-aa peptides resulted in small effect on kinetic parameters. Ionic strength higher than physiologic depressed thrombin action on the 25-aa peptides. Addition of ligands of the exosite 1 negatively modulated the catalysis of 25-aa substrates. In conclusion, we succeeded to mimic and study in real time, using these synthetic peptides, the influence of ligand binding to exosite 1 on thrombin activity. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|