The goal of this research is to produce Metabolic Control Analysis (MCA) and Flux Balance Analysis (FBA) models of the sialic acid biosynthetic system in order to identify control points that govern the production of specific forms of sialic acid that modulate particular cellular behaviors. The project will utilize an integrative approach where a model will be constructed, experimental data will be obtained to test the robustness of the model, and the model will be further fine-tuned to test its response to external stimuli. More specifically, the MCA model will be constructed based on known, or estimated, enzyme kinetic parameters. Metabolic flux through the sialic acid pathway will then be controlled through the use of deuterium-labeled metabolic substrate, which will allow small-molecule metabolite production, expression of the biosynthetic genes, and product formation to be measured quantitatively by mass spectroscopy, real-time PCR, and immunological methods, respectively. These data will be used to refine the models. Finally, the models will be validated by subjecting them to stimuli known to perturb sialic acid metabolism; the results of the experimentally-obtained and modeled data will be compared to test the robustness of the models.
