The main goals of the stay proposed are as follows: 1. Contribute to better understanding of mechanism of substrate oxidations by cytochromes P-450 and peroxidases. The experimental work implicates to learn to isolate particular isoenzymes of cytochrome P-450 and the respective reductase and to purify the peroxidase, then, to learn how to perform experiments with the reconstituted P-450 systems, how to analyze the reaction mixtures by the HPLC and how to identify the metabolites with the GC/MS. For evaluation of possible mechanisms, the use of isotopically labelled compounds is needed. I would like to learn the skills and conceptual approaches of measuring the kinetic isotope effects on enzyme-catalyzed reactions. In summary, I would like to learn how to perform experiments on metabolism of various (labeled or unlabeled) substrates with defined systems of cytochromes P-450 and peroxidases, how to analyze the reaction mixtures with the advanced techniques and how to evaluate the results using isotope effects. 2. As I work currently in the field of the metabolism of foreign compounds, I expect to use the approaches and methods for my own work in my home institution. I wish to share the knowledge gained in the U.S. also with my graduate students at the Charles University. I expect that during my first stay I will learn the techbioquesand that I will be able to perform first experiments; during the second and the third stay I would like to pursue the experiments and to finalize the project in the form of a research communication.
