Kinetic study of the activation process of frog epidermis pro-tyrosinase by trypsin.
1. The rate of tyrosinase formation has been calculated by coupling the activatory process of frog epidermis pro-tyrosinase by trypsin to the oxidation of L-DOPA to dopachrome. Under certain conditions ([trypsin]/[pro-tyrosinase] greater than or equal to 300), the lag period of the coupled reactions, tau, is independent of trypsin concentration. 2. The specific rate constant of tyrosinase formation at different temperatures has been calculated, ranging from 0.025 sec-1, at 5 degrees C to 0.248 sec-1, at 30 degrees C. 3. Thermodynamic parameters of the activatory process (delta G not equal to = + 18.5 kcal/mol; delta H not equal to = + 14.8 kcal/mol; delta S not equal to = -12.4 e.u.; Ea = + 15.3 kcal/mol), have been determined by the study of the system at different temperatures. These values are characteristic for a normal chemical reaction. 4. From these kinetic data, the order of products formation in the proteolytic step, can be determined, active tyrosinase being the last product released.
D000818:Animals; D004156:Catechol Oxidase / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004792:Enzyme Precursors / Q000378:metabolism*; D004817:Epidermis / Q000201:enzymology*; D007700:Kinetics; D008956:Models, Chemical; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D011893:Rana esculenta; D014357:Trypsin / Q000494:pharmacology*
D004792:Enzyme Precursors; D004156:Catechol Oxidase; D014442:Monophenol Monooxygenase; D014357:Trypsin