RUI: Microelectrophoretic and Statistical Tools for Studies of Kinase- and ROS-Based Signaling in Dictyostelium discoideum

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Living organisms use a diversity of molecules to communicate information within and between cells, and scientists are still investigating the interactions between these molecules. This project will produce simultaneous measurements of both large proteins and small reactive, oxygen-containing molecules in groups of cells and in individual cells to better understand the interactions between these molecular signals. The project will also develop mathematical tools to analyze the data collected from these experiments and make these tools available to researchers doing similar work. Finally, this project will engage undergraduates and high school students in interdisciplinary research involving cell signaling, separations chemistry, microfluidics, and mathematics.The amoeba Dictyostelium discoideum undergoes a social life cycle, in which individual cells aggregate and differentiate to form a superorganism. Cell signaling during this process involves both protein kinases and reactive oxygen species, but the relationship between these interacting signaling nodes remains unclear. Through microelectrophoretic separations of peptide substrates and fluorogenic indicators, this project will simultaneously measure protein kinase B activity and superoxide concentrations in cells. In addition to characterizing these molecules in cell lysates, this project will also produce single-cell measurements using custom-designed microfluidic devices and novel mathematical strategies for data analysis.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.