Kinetic analysis of the cleavage of human protease-activated receptor-1 / 2 / 3 and 4 using quenched-fluorescent peptide substrates.
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, k(cat) / K(m) values were determined.
Fox
Mark T
MT
Centre for Peptide and Protein Engineering, School of Biology and Biochemistry, Medical Biology Centre, The Queen's University of Belfast, 97, Lisburn Road, Belfast, BT9 7BL, Northern Ireland. m.t.fox@qub.ac.uk
Greer
Brett
B
Lawson
Jill
J
Healy
Adrienne
A
Harriott
Patrick
P
D007700:Kinetics; D011499:Protein Processing, Post-Translational; D044463:Receptor, PAR-1; D044464:Receptor, PAR-2; D018179:Receptors, Thrombin / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013917:Thrombin / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D044463:Receptor, PAR-1; D044464:Receptor, PAR-2; D018179:Receptors, Thrombin; C105295:protease-activated receptor 3; D014357:Trypsin; D013917:Thrombin; C112908:protease-activated receptor 4