Comparative Analysis of Stability-Based Profiling Techniques and Their Application to the Characterization of Drug Targets and Disease Phenotypes

Morgan A. Bailey, Ph.D. Candidate Michael C. Fitzgerald Ph.D., Advisor Abstract: The advancement of mass spectrometry-based protein stability profiling measurements within the past twenty years has led to the development of a suite of approaches that enables the evaluation of protein folding stability on a broad range of biological mixtures with varying complexity. These approaches include chemical and thermal denaturation approaches (SPROX and TPP, respectively) as well as proteolysis strategies such as limited proteolysis (LiP) and pulse proteolysis (PP) which have all been extensively used and evaluated for small molecule protein target discovery applications. However, the capabilities of these methods have yet to be fully evaluated in the characterization of disease phenotypes and other biological events such as post-translational modifications and RNA-protein interactions. A major focus of the work included in this dissertation has been the comparative analysis of the above techniques for the analysis of biological phenotypes. The application and comparative analysis of the above techniques to the characterization of RNA-protein interactions is also described.