Kinetic Analysis of the Cleavage of Human Protease-Activated Receptor-1 / 2 / 3 and 4 Using Quenched-Fluorescent Peptide Substrates

Title: Kinetic Analysis of the Cleavage of Human Protease-Activated Receptor-1 / 2 / 3 and 4 Using Quenched-Fluorescent Peptide Substrates

Volume: 9 Issue: 5

Author(s): Mark T. Fox, Brett Greer, Jill Lawson, Adrienne Healy and Patrick Harriott

Affiliation:

Keywords: Enzymology, serine proteases, progress-curve kinetics, fluorescence, 2-aminobenzoyl (Abz), 2,4-dinitrophenyl (Dnp), fluorescence resonance, G-protein-coupled, seven-transmembrane, domain receptor

Abstract: Protease-activated receptors [PARs] are a family of G-protein-coupled seven-transmembrane domain receptors that are activated by proteolytic cleavage of their amino-terminal exodomain. To characterize the cleavage rate of human PAR-1 / 2 / 3 and 4 by trypsin and thrombin, four synthetic quenched-fluorescent peptide substrates have been synthesized. Each substrate consisted of a ten-residue peptide spanning the receptor activation cleavage site and using progress-curve kinetics, kcat / Km values were determined.

Cite this article as:

Fox T. Mark, Greer Brett, Lawson Jill, Healy Adrienne and Harriott Patrick, Kinetic Analysis of the Cleavage of Human Protease-Activated Receptor-1 / 2 / 3 and 4 Using Quenched-Fluorescent Peptide Substrates, Protein & Peptide Letters 2002; 9 (5) . https://dx.doi.org/10.2174/0929866023408490