Structure and function of glycolytic enzymes and their cellular regulation

PROJECT SUMMARY Glucose metabolism plays a crucial role in cellular homeostasis and is central to diseases like cancer, diabetes, and neurodegenerative disorders. Despite extensive biochemical and enzymological characterization, the cell biology of many metabolic enzymes remains poorly understood. Targeting glycolytic enzymes in diseases in which they are dysregulated, such as cancer, diabetes, and inflammation, is a currently proposed therapeutic strategy. However, this strategy requires understanding how glycolytic enzymes are regulated in time and space, a process that remains unresolved. Phosphofructokinase-1 (PFK1), a key regulatory enzyme of glycolysis, is controlled through complex mechanisms, but its spatial and temporal dynamics in cells and role in disease-related metabolic reprogramming remain unclear. This proposal will determine the regulation of PFK1 in vitro and in cells. Specifically, we will examine the how the liver isoform of PFK1 assembles into filaments and how this regulates glucose metabolism in hepatocytes and how a glycolytic metabolon regulates the migration of breast cancer cells. We will also determine how the carboxy terminal tail domain of PFK1 stabilizes the inactive conformation and how its inhibitory effects are relieved in response to metabolic and hormonal signaling. By employing high-resolution structural analyses, cellular assays, and innovative molecular tools, the project seeks to provide a comprehensive understanding of the spatial and temporal regulation of PFKL. Our findings will advance the field of cell metabolism and cell biology, will provide insights into their dysregulation in disease, and may aid in the development of novel therapeutic interventions.