NSF Award
9020248
The principal goal of this project is to obtain information on the structure and function of the yeast Saccharomyces cerevisiae mitochondrial ATP synthetase as it relates to the mechanism of ATP synthesis. Yeast is an excellent system in part because of the established genetics including homologous recombination and a transformation system, yeast is a facultative anaerobe and thus mutations in the ATP synthetase can be isolated and because of the high amount of homology that the yeast ATP synthetase has with other organisms. The objectives of this work are two-fold: to complete the biochemical analysis of the yeast mitochondrial ATPase and to study a portion of the active site of the ATPase (the glycine rich loop region) by saturation mutagenesis followed by biochemical analysis. Previous evidence from this and other laboratories suggests that this loop is essential for activity of the enzyme and conformational changes by this loop are instrumental in the kinetic responses of the ATPase if not for the allosteric properties of the enzymes. This project will provide a further understanding into how the mitochondrial ATP synthetase utilizes the energy from the proton gradient to make ATP.
