Kinetic study of prolidase activity in erythrocytes against different substrates using capillary electrophoresis with electrochemiluminescence detection.
Capillary electrophoresis (CE) with electrochemiluminescence (ECL) detection was used to explore the kinetics of the enzymatic reaction. The different effects of reaction conditions including the concentration of Mn(2+), incubation temperature and pH on prolidase (PLD, EC 3.4.13.9) activity in erythrocyte lysates against three different substrates, Gly-Pro, Val-Pro and Leu-Pro were investigated. Also, the effects of colchicine which can prevent or delay cancer of liver on the PLD activity were studied. For Gly-Pro and Leu-Pro, colchicine enhanced PLD activity, while for the other substrate Val-Pro, it had a slight inhibiting effect on prolidase activity. Kinetic study of colchicine on prolidase activity in erythrocytes indicated different effects for different substrates with the addition of colchicine. Compared with some other methods, CE coupled with electrochemiluminescence detection showed its merits of simplicity and efficiency on the bioanalysis.
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Graduate School of the Chinese Acadamy of Sciences, Changchun, Jilin 130022, China.
Lingling
L
Zhou
Jipei
J
Yuan
Jianyuan
J
Yin
Erkang
E
Wang
D004150:Dipeptidases / Q000378:metabolism*; D019075:Electrophoresis, Capillary / Q000379:methods*; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008163:Luminescent Measurements / Q000379:methods*; D009842:Oligopeptides / Q000378:metabolism; D015203:Reproducibility of Results; D013379:Substrate Specificity; D013696:Temperature
D009842:Oligopeptides; D004150:Dipeptidases; C022449:proline dipeptidase