Kinetic study of carboxypeptidase P-catalyzed reaction. Pressure and temperature dependence of kinetic parameters.
Detailed kinetic analyses of carboxypeptidase P-catalyzed reactions were carried out spectrophotometrically using 3-(2-furyl)acryloyl-acylated peptide substrates. The maximum kcat/Km was observed at around pH 3.5 for the synthetic peptide substrates. The kcat/Km value decreased with increasing pH, with an apparent pKa value of 4.43. However, the maximum kcat was observed at neutral pH (pH congruent to 6) and the pKa was 4.49. These apparently different pH profiles for kcat/Km and kcat of this enzyme were due to the decreasing Km value in the acid pH region. The pressure and temperature dependences of these kinetic parameters were also measured. N-Benzoylglycyl-L-phenyllactate (Bz-Gly-OPhLac) gave dependences similar to those of the peptide substrate, suggesting that there is no distinct difference in the catalytic mechanism between the peptide and the ester hydrolyses.
D002268:Carboxypeptidases / Q000378:metabolism*; D043422:Carboxypeptidases A; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011485:Protein Binding; D013379:Substrate Specificity; D013816:Thermodynamics
C025427:carboxypeptidase P; D002268:Carboxypeptidases; C022633:serine carboxypeptidase; D043422:Carboxypeptidases A