NSF Award
8703618
The goal of this project to gain an understanding of the structure and function of mitochondrial ATP synthase. Specifically, molecular genetic approaches will be applied to an investigation of two key peptides of the oligomeric enzyme from yeast: 1) the beta subunit of the F1-ATPase, and 2) the oligomycin sensitive conferring peptide (OSCP) that links F1 with F0 domain. The beta subunit is thought to be at least a part of the catalytic site for ATP synthesis. The role of specific amino acids in catalysis or binding will be evaluated by site directed mutagenesis using oligonucleotides and by random mutagenesis using chemical methodology. The mutations will be studied by introducing the mutated genes into a beta subunit deficient yeast strain determining its effect on function. For the OSCP unit the gene will be cloned, sequenced and studied in a similar manner as is planned for the beta subunit. The results of the research should enhance our understanding of how ATP, the key compound involved in transduction of energy in biological systems, is synthesized.
