A kinetic study of the suicide inactivation of an enzyme measured through coupling reactions. Application to the suicide inactivation of tyrosinase.
A systematic procedure for the kinetic study of reaction mechanisms with enzyme inactivation induced by a suicide substrate in the presence or in the absence of an auxiliary substrate, when the enzyme activity is measured through coupling reactions, enzymically catalysed or not, was developed and analysed by using the transient-phase approach. The methodology is established to determine the parameters and kinetic constants corresponding to the enzyme suicide inactivation and the coupling reactions. This approach is illustrated by a study of the suicide inactivation of tyrosinase by catechol in the presence of L-proline. Treatment of the experimental data was carried out by non-linear regression.
Escribano
J
J
Departamento de Bioquimica y Biologia Molecular, Universidad de Murcia, Spain.
Tudela
J
J
Garcia-Carmona
F
F
Garcia-Canovas
F
F
D004156:Catechol Oxidase / Q000037:antagonists & inhibitors*; D002396:Catechols / Q000378:metabolism; D007700:Kinetics; D014442:Monophenol Monooxygenase / Q000037:antagonists & inhibitors*; D011392:Proline / Q000378:metabolism
D002396:Catechols; D011392:Proline; D004156:Catechol Oxidase; D014442:Monophenol Monooxygenase