Artigo
Investigation of Thrombin Activity with PAR 1-based Fluorogenic Peptides
Fecha
2013-10-01Registro en:
Protein And Peptide Letters. Sharjah: Bentham Science Publ Ltd, v. 20, n. 10, p. 1129-1135, 2013.
0929-8665
10.2174/09298665113209990001
WOS:000323042100007
Autor
Vieira, Saulo Martins
Reis, Flavia Coelho Garcia dos
Geraldo, Reinaldo Barros
Dutra, Denis Luis da Silva
Juliano, Luiz [UNIFESP]
Juliano, Maria Aparecida [UNIFESP]
Mignaco, Julio Alberto
Zingali, Russolina Benedeta
Institución
Resumen
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 P-3' 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.