Structure Function Studies of Cytochrome c

点击翻译按钮获取中文摘要

This interdisciplinary project supported by the Inorganic, Bioinorganic and Organometallic Chemistry Program in the Mathematical and Physical Sciences Directorate and the Biophysics program in the Biological Sciences Directorate combines the expertise of chemists and molecular biologists in the elucidation of structure function relationships in cytochrome c. The aims of the research are: 1) to examine, in vitro, the effects of mutations within the heme environment on the redox potential and electron transfer reactivity of the cytochrome, and to compare the effects observed in vitro with the in vivo reactivity of the cytochrome, and 2) to determine the stabilities of the mutant cytochromes using both kinetic and thermodynamic methods with the ultimate objective of determining how protein stability can be linked to the level of protein expression of the cell. Recombinant DNA procedures will be used to obtain the mutant cytochromes from the yeast Saccharomyces cerevisiae that will be used in these studies. Estimation of the amount of cyctochrome present in cells will be done using the spectral response of the heme chromaphore. %%% The objective of this research is to further our understanding of the ways in which variations in the structure of biomolecules, and that of metalloproteins in particular, affect their biological function. Because of the difficulty of making direct observations on biological molecules in vivo, studies will initially be made in vitro and then behavior that can be studied in vivo, such as the rate of protein production by the cell, will be correlated with the in vitro measurements.