PROTEIN STRUCTURE, STABILITY AND DYNAMICS IN CELLS AND CELL-LIKE ENVIRONMENTS Public Deposited
- Last Modified
- March 19, 2019
- Creator
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Smith, Austin
- Affiliation: College of Arts and Sciences, Department of Chemistry
- Abstract
- The intracellular milieu is filled with small molecules, nucleic acids, lipids and proteins. Theories have attempted to explain how macromolecules react to this environment for over 30 years. Recent experiment-based studies have shown that protein stability and dynamics are altered in this environment. I used the loop of chymotrypsin inhibitor 2 and two unfolded proteins (α-synuclein and FlgM) to show that the crowded cellular matrix does not necessarily cause structuring of these dynamic regions. Most importantly, I have shown the thermodynamic and mechanistic basis for how protein stability is changed in the cellular environment. To do this I use a marginally stable globular protein (an isolated SH3 domain) to measure stability, dynamics, and folding rates in cells and cell-like environments. Proteins are enthalpically destabilized in cells. The destabilization arises from charge-charge interactions of the cellular environment with the unfolded ensemble of the protein. These interactions also slow folding of the protein. This work will allow creation of a more complete picture of protein thermodynamics inside the cell. Furthermore, the SH3 domain is amenable to studying in vitro protein stability over a broad range of pH values, and allows acquisition of folding and unfolding rates with a variety of crowders. Future efforts will facilitate a better understanding of surface charge interactions and will allow elucidation of a crowder’s interaction with the transition state.
- Date of publication
- August 2015
- Keyword
- Subject
- DOI
- Identifier
- Resource type
- Rights statement
- In Copyright
- Advisor
- Berkowitz, Max
- Thompson, Nancy
- Spremulli, Linda
- Pielak, Gary J.
- Lee, Andrew
- Degree
- Doctor of Philosophy
- Degree granting institution
- University of North Carolina at Chapel Hill Graduate School
- Graduation year
- 2015
- Language
- Publisher
- Place of publication
- Chapel Hill, NC
- Access
- There are no restrictions to this item.
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This work has no parents.
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