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Protein & Peptide Letters

Editor-in-Chief

ISSN (Print): 0929-8665
ISSN (Online): 1875-5305

Investigation of Thrombin Activity with PAR 1-based Fluorogenic Peptides

Author(s): Saulo Martins Vieira, Flavia Garcia dos Reis, Reinaldo Geraldo, Denis Luis da Silva Dutra, Luiz Juliano, Maria Aparecida Julianod, Julio Alberto Mignaco and Russolina Benedeta Zingali

Volume 20, Issue 10, 2013

Page: [1129 - 1135] Pages: 7

DOI: 10.2174/09298665113209990001

Price: $65

Abstract

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.

Keywords: Enzyme activity, exosite 1, fluorogenic peptides, PAR 1 and thrombin.


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