Date of Award
Doctor of Philosophy (PhD)
Mass spectrometry (MS) is a powerful tool for analyzing biomolecules. In this dissertation, we present development and applications of carbene reagents for protein footprinting and hydrogen/deuterium exchange (HDX) for characterization of a protein complex that is important for nuclear transportation of an Ebola virus protein. In Chapter 1, we describe the basic principles and detailed configurations of Photochemical Oxidation of Proteins (FPOP). FPOP utilizes hydroxyl radicals to probe protein higher order structure. To expand the radical reagents used for FPOP, we explored the feasibility of generating carbenes by laser photolysis of diazirines in our FPOP system for protein footprinting in Chapters 2 and 3. In Chapter 4, we took advantage of one diazirine reagent and studied the structural differences of apolipoprotein E3 and E4, and their interactions with a small molecule structural corrector. In chapter 5, we described a novel method to combined carbene footprinting with hydroxyl radical footprinting in a two-laser FPOP platform. We also applied several other MS-based methods in Chapter 6 to characterize protein complexes that are involved in nuclear transportation of Ebola VP24 protein.
Chair and Committee
Michael L. Gross
Alexander Barnes, Robert Blankenship, Daisy Leung, John-Stephen Taylor,
Zhang, Bojie, "Mass Spectrometry Methods to Characterize Protein Higher Order Structure - From Ebola to ApoE" (2019). Arts & Sciences Electronic Theses and Dissertations. 1820.
Available for download on Monday, May 15, 2119