Darrah, Kristie
(2021)
Chemical Tools to Control Protein Expression, Function, and Degradation.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
Abstract
The precise timing and localization of the complex interactions among small molecules and biomacromolecules during cellular biological processes directly correlate to larger changes that can have long withstanding impacts on overall human health. Chemical tools to discreetly investigate the individual contribution of key cellular events or signaling pathways are vital to uncovering answers to many of the questions within the realm of chemical biology. Herein, I describe my contributions towards developing and applying oligonucleotide- and small molecule- based chemical tools to the fields of developmental biology, sulfotransferase biology, and targeted protein degradation.
Using morpholino oligonucleotide antisense agents, I have developed conditionally activated, cyclic morpholino reagents responsive to enzyme catalysis or small molecule treatment and have applied them in controlling endogenous gene expression in zebrafish embryos. In combination with current optically controlled technologies, these reagents can be used to silence expression of individual genes within multi-gene networks. I have also synthesized the first synthetic, allosteric small molecule inhibitor of any sulfotransferase enzyme. This work uncovered the sulfotransferase active site cap as an entirely novel allosteric pocket that can be exploited for small molecule regulation. The inhibitors presented in this work have laid the groundwork for a potentially novel approach to treating major depression disorder. Finally, I have contributed to implementing a broadly applicable approach for the optical activation of small molecule-induced protein degradation. By utilizing a general photocaging strategy, the approach presented in this work permits spatiotemporal control over PROTAC-mediated protein degradation.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
|
| ETD Committee: |
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| Date: |
8 October 2021 |
| Date Type: |
Publication |
| Defense Date: |
28 July 2021 |
| Approval Date: |
8 October 2021 |
| Submission Date: |
10 August 2021 |
| Access Restriction: |
2 year -- Restrict access to University of Pittsburgh for a period of 2 years. |
| Number of Pages: |
348 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
Dietrich School of Arts and Sciences > Chemistry |
| Degree: |
PhD - Doctor of Philosophy |
| Thesis Type: |
Doctoral Dissertation |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
chemical biology, morpholino, conditional control, antisense agents, sulfotransferase, small molecule inhibitors, targeted protein degradation |
| Additional Information: |
Erratum applied for the following changes:
Figure 1-19 contained images that were repeated in Figure 1-40.Additionally, three of the reagent recipes in the Expanded Methodschapter were listed wrong. The new version of this ETD PDF containsthe correct images that correspond to the data presented in the Figure 1-19 and the corrected reagent recipes. |
| Date Deposited: |
08 Oct 2021 20:26 |
| Last Modified: |
08 Nov 2023 13:35 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/41633 |
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