This item is under embargo and not available online per the author's request. For access information, please visit http://libanswers.wustl.edu/faq/5640.

ORCID

http://orcid.org/0000-0001-7690-9796

Date of Award

Summer 8-15-2019

Author's School

Graduate School of Arts and Sciences

Author's Department

Chemistry

Degree Name

Doctor of Philosophy (PhD)

Degree Type

Dissertation

Abstract

Nuclear magnetic resonance (NMR) is a nondestructive experimental technique that often relies on radio frequency and large magnetic fields to investigate molecular structure and dynamics. The technique is well suited to study complex biological systems; however, universal application, to all samples, is hindered by low sensitivity. While several methods to increase NMR sensitivity currently exist, dynamic nuclear polarization (DNP) is one method gaining increased attention in the solid-state NMR community. This dissertation describes the development of DNP instrumentation for magic-angle spinning (MAS) NMR, including an efficient counterflow liquid nitrogen heat exchanger and novel spherical rotors. Furthermore, contained herein is the development of in-cell DNP. For the first time, DNP enhanced NMR signals from the intracellular regions of intact human cells are confirmed through the application of new fluorescent trimodal polarizing agents. The combination of fluorescent microscopy and fluorescence activated cell sorting with MAS DNP NMR provides a unique analytical platform to investigate intracellular molecular structures and dynamics. The in-cell DNP techniques developed in this research are expanded to investigate activation of HIV virus from latently infected human cell lines by NMR. Lastly, novel combinations of latency reversing agents, typically utilized in the “shock and kill” approach to HIV eradication to potently activate HIV from its latent state, are presented.

Language

English (en)

Chair and Committee

Alexander B. Barnes

Committee Members

George B. Kyei, Sophia E. Hayes, Jacob Schaefer, Timothy A. Wencewicz,

Available for download on Wednesday, November 04, 2020

Share

COinS