Dissertation
DEVELOPMENT AND APPLICATION OF ANALYTICAL AND CHEMICAL BIOLOGY APPROACHES TO UNDERSTAND PHYSIOLOGICAL RESPONSES TO EXTERNAL EXPOSURES
Doctor of Philosophy (PhD), Washington State University
01/2020
Handle:
https://hdl.handle.net/2376/111917
Abstract
Activity-based probes allow for the identification of functionally similar proteins while providing a measure of their activity. They are central to each chapter in this dissertation, which focuses on the development and application of an activity-based probe and analytical methods to understand physiological responses to external exposures. Chapter 1 introduces the concept of activity-based protein profiling, the design of activity-based probes, and methods for analyzing probe-targeted proteins that leads the reader to an in-depth understanding of activity-based protein profiling. Chapter 2 includes the design, synthesis, validation, and application of a glutathione-mimicking activity-based probe with excellent selectivity for glutathione-binding proteins such as the glutathione S-transferase (GST) family of enzymes. The capability of the probe to distinguish differential activity between different GST enzymes was practically demonstrated in the identification of a contrasting response of murine GSTs to the carcinogen benzo[a]pyrene. Chapter 3 discusses a new statistical application for advancing the identification of differentially abundant proteins between sample groups, a task commonly performed to identify activity-based probe protein targets and detect their differential activity. Comparison with standard methods revealed ours outperforms others at detecting differentially abundant proteins, particularly those with minimal differential abundance. Chapter 4 discusses the analysis and interpretation of complex activity-based protein profiling and multi-omics data originating from the gut microbiomes of mice subjected to high or low fiber diets. Biological interpretation of analytical results led to the identification of critical aspects of mucosal layer degradation in response to fiber deprivation. Finally, chapter 5 concludes with a summary of findings and contributions to the scientific community that advance our understanding of physiological responses to external exposures.
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Details
- Title
- DEVELOPMENT AND APPLICATION OF ANALYTICAL AND CHEMICAL BIOLOGY APPROACHES TO UNDERSTAND PHYSIOLOGICAL RESPONSES TO EXTERNAL EXPOSURES
- Creators
- Bryan James Killinger
- Contributors
- Aaron T Wright (Advisor)Alla Kostyukova (Committee Member)Wenji Dong (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Chemical Engineering and Bioengineering, School of
- Theses and Dissertations
- Doctor of Philosophy (PhD), Washington State University
- Number of pages
- 131
- Identifiers
- 99900581810601842
- Language
- English
- Resource Type
- Dissertation