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ORCID

http://orcid.org/0000-0002-3139-4159

Date of Award

Spring 5-15-2019

Author's School

Graduate School of Arts and Sciences

Author's Department

Chemistry

Degree Name

Doctor of Philosophy (PhD)

Degree Type

Dissertation

Abstract

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.

Language

English (en)

Chair and Committee

Michael L. Gross

Committee Members

Alexander Barnes, Robert Blankenship, Daisy Leung, John-Stephen Taylor,

Comments

Permanent URL: https://doi.org/10.7936/7xah-wx61

Available for download on Monday, May 15, 2119

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