Date of Award
Doctor of Philosophy (PhD)
The dissertation will be solely focused on using mass spectrometry to characterize protein high order structures (HOS), it emphasizes the use of hydroxyl radical footprinting (FPOP) coupled to bottom-up MS approach. A detailed background information about FPOP, and the corresponding method developments as well as applications will be covered.
The first chapter will be a comprehensive review regarding the FPOP. Following this, chapter 2, 3, and 4 will be focused on the method developments. Chapter 2 describes an isotope dilution GC-MS method to quantitate OH radicals in FPOP; chapter 3 describes the incorporation of Leu-enkephalin as reporter peptide for a more quantitative FPOP platform; and chapter 4 introduces how R-programming can facilitate the MS-based structural proteomics. After this, chapter 5, 6, and 7 are mainly about the applications of FPOP to characterize the proteins. Chapter 5 talks about using FPOP to localize the dimer dissociation and local unfolding of G93A SOD1; chapter 6 describes how FPOP can be used to characterize an intrinsically disordered tail of EGF receptor protein; and chapter 7 demonstrates the feasibility of a marriage of FPOP and Nanodiscs to study the membrane-associated KRAS protein.
Chair and Committee
Gaya Amarasinghe, Robert Blankenship, Dewey Holten, Jay Ponder,
Niu, Ben, "Mass Spectrometry-based Structural Proteomics: Methodology and Application of Fast Photochemical Oxidation of Proteins (FPOP)" (2017). Arts & Sciences Electronic Theses and Dissertations. 1238.