Date of Award
Doctor of Philosophy
Michael D. Best
Shawn R. Campagna, David M. Jenkins, Todd B. Reynolds
Lipids are biologically important molecules as they regulate a litany of key cellular processes that, when aberrant, result in disease. Since lipids control these regulation pathways, it is imperative to understand their properties and exploit them for use in advancing therapeutic strategies. The complex environments in which lipids exist have historically complicated their understanding, but recent advancements have facilitated this effort. In the chapters that follow, we have used synthetic lipid probes to investigate the role of signaling lipids as site-specific ligands in the binding of protein effectors and developed a high-throughput platform that can be used to rapidly classify protein-binding partners to phosphoinositide lipids. Using microarray analysis, we have achieved high-throughput binding results for the Akt-PH domain to phosphatidylinositol-(3,4,5)-trisphosphate. In addition, preliminary results from an investigation of how multivalency affects profilin binding to phosphatidylinositol-(4,5)-bisphosphate are discussed. We have also developed the first known example of liposome modification via copper-free click chemistry as an ideal bioorthogonal approach to intact membrane derivatization towards targeting applications. Using a microplate surface-based immobilization technique, we have reported the derivatization of intact azido-liposomes with a biotinylated cyclooctyne by copper-free click chemistry and performed various control reactions to ensure labeling is specific and reliable. Additionally, we have reported initial findings for the advancement of this liposome immobilization strategy to be performed in high-throughput fashion via microarray. In the final chapter, we briefly discuss our involvement in a few collaborative efforts including the optimization of a non-lipid autotaxin inhibitor, synthesis of porphyrin-lipid conjugates for use in photocurrent generation systems, and synthesis of several rhodamine dyes for use in surface enhanced hyper Raman spectroscopy (SEHRS).
Bostic, Heidi Elizabeth, "Development and Application of Synthetic Lipid Probes to Study and Mimic Lipid Function. " PhD diss., University of Tennessee, 2012.