Spatiotemporal dynamics of protein kinase C signaling

Despite many advances in contemporary pharmacology and cell biology, there still exists an epistemological chasm between data gathered biochemically and data gathered from whole animal or tissue. In complex systems of signal transduction, such as for Protein Kinase C (PKC), the lack of precise information at the cellular level can preclude meaningful integration of biochemical and animal data into comprehensive understanding of a signaling system. This dissertation extends knowledge gained from previous PKC research to live cell experiments using technologies based upon A. Victoria Green Fluorescent Protein. Specifically, this dissertation shows that genetically encoded fluorescent reporters for PKC conformational changes, lipid raft microdomains, and PKC substrate phosphorylation provide molecular detail to PKC function in living cells. The results presented here show a remarkable fidelity of PKC signal transduction to dynamic regulation, and suggest that such methods may be applicable to a wide variety of kinase signals in living cells. We describe PKC as a signal integrator that allows cells to process rapidly changing environmental cues with high temporal precision, providing cells with a mechanism for high-content information in the form of dynamic phosphorylation.