The purpose of this project is to demonstrate that, using isothermal titration calorimetry, I can measure the typical metabolic activity of Jurkat cells under normal conditions and also use inhibitors to determine the proportion of energy these cells obtain from anaerobic and aerobic metabolic pathways. We can then treat with chemotherapeutics and compare to see how these compounds affect aerobic and anaerobic metabolism. ITC is a method usually used to measure ligand-receptor binding interactions, enzyme kinetics, and reaction enthalpy, but we propose to use it as a novel tool for use in metabolic analysis. This can be done by placing cells or chemicals into the reaction vessel, choosing a temperature for the experiment, and setting up injections of your compound/enzyme of interest. In this case, as the cells metabolize they produce heat, and the amount of energy that the calorimeter puts out to maintain a constant solution temperature is decreased. This produces a graph that represents the activity of the cell over time. We postulate that using this method we will be able to determine the typical average heat rate of each individual cell as well as find the proportion of energy the cells derive from aerobic and anaerobic processes by isolating these processes using inhibitors.
