Investigating structural and stability characteristics of native-like proteins using ion mobility-mass spectrometry (IM-MS) in positive- and negative-ion mode.

Edwards, Alexis N., 1997-

Investigating structural and stability characteristics of native-like proteins using ion mobility-mass spectrometry (IM-MS) in positive- and negative-ion mode.

dc.contributor.advisor	Gallagher, Elyssia S.
dc.creator	Edwards, Alexis N., 1997-
dc.date.accessioned	2024-07-30T12:42:24Z
dc.date.available	2024-07-30T12:42:24Z
dc.date.created	2023-12
dc.date.issued	2023-12
dc.date.submitted	December 2023
dc.date.updated	2024-07-30T12:42:24Z
dc.description.abstract	Native ion mobility-mass spectrometry (IMS-MS) is widely used to investigate the structure and stability of biologically relevant proteins and protein complexes. The key goal, and arguably biggest attraction, of native IMS-MS is maintaining the noncovalent interactions responsible for stabilizing the three-dimensional structures of a protein when it is introduced to the gas-phase. By maintaining and investigating a folded protein’s gas-phase structure, one can learn the stoichiometry, subunit composition, and binding constants. The focus of this dissertation is investigating several fundamental aspects of native IMS-MS for analysis of protein and protein complexes. In Chapter One, the importance of studying protein structure and how native IMSMS is a valuable tool to do so is discussed in detail. In Chapter Two, we investigate the sources of uncertainty present in native IMS-MS analysis. There are several sources of uncertainty in IMS analysis that are not routinely monitored or regularly reported, particularly for Traveling-Wave Ion Mobility Spectrometry (TWIMS) calibration, so we developed an error propagation method that quantifies some of this uncertainty. Quantifying this uncertainty allows for a more robust comparison of protein gas-phase structures. Chapter Three focuses on how a protein’s net-charge in solution affects its overall structure and stability in the gas-phase. We were able to demonstrate that some proteins are more stable in the gas-phase when analyzed in the ion mode matching their solution-phase charge, thus indicating the ionization process can cause changes to a protein’s stability even when it does not change the overall three-dimensional structure. Lastly, in Chapter Four we use native IMS-MS to determine the structure and stability of a biologically relevant ternary enzymatic complex, highlighting the utility of native IMSMS for characterization of bound and unbound enzymatic samples. Together, this work broadens the native IMS-MS communities’ understanding of several aspects of nativelike protein analysis via IMS-MS while also showcasing the utility of such techniques for the analysis of biologically relevant samples.
dc.format.mimetype	application/pdf
dc.identifier.uri
dc.identifier.uri	https://hdl.handle.net/2104/12859
dc.language.iso	English
dc.rights.accessrights	No access – contact librarywebmaster@baylor.edu
dc.title	Investigating structural and stability characteristics of native-like proteins using ion mobility-mass spectrometry (IM-MS) in positive- and negative-ion mode.
dc.type	Thesis
dc.type.material	text
local.embargo.lift	2025-12-01
local.embargo.terms	2025-12-01
thesis.degree.department	Baylor University. Dept. of Chemistry & Biochemistry.
thesis.degree.grantor	Baylor University
thesis.degree.name	Ph.D.
thesis.degree.program	Chemistry
thesis.degree.school	Baylor University