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

活着的生物体使用不同的分子在细胞内和细胞之间交流信息，科学家们仍在研究这些分子之间的相互作用。该项目将同时测量细胞组和单个细胞中的大蛋白质和小的活性含氧分子，以更好地了解这些分子信号之间的相互作用。该项目还将开发数学工具来分析从这些实验中收集的数据，并将这些工具提供给从事类似工作的研究人员。最后，这个项目将吸引本科生和高中生进行跨学科的研究，涉及细胞信号、分离化学、微流体学和数学。盘状阿米巴经历了一个社会生命周期，在这个社会周期中，单个细胞聚集和分化形成一个超级有机体。在这一过程中，细胞信号转导同时涉及蛋白激酶和活性氧物种，但这些相互作用的信号转导节点之间的关系尚不清楚。通过多肽底物和荧光指示剂的微电泳法分离，该项目将同时测量细胞中蛋白激酶B的活性和超氧化物浓度。除了在细胞裂解产物中表征这些分子，该项目还将使用定制设计的微流控设备和用于数据分析的新颖数学策略来产生单细胞测量。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.