Abstract

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.

Committee Chair

Michael Gross

Committee Members

Gaya Amarasinghe, Robert Blankenship, Dewey Holten, Jay Ponder,

Comments

Permanent URL: https://doi.org/10.7936/K7P55MXM

Degree

Doctor of Philosophy (PhD)

Author's Department

Chemistry

Author's School

Graduate School of Arts and Sciences

Document Type

Dissertation

Date of Award

Summer 8-15-2017

Language

English (en)

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