Gaining insight into electron capture dissociation mass spectrometry of peptides and proteins Public Deposited

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  • March 20, 2019
Creator
  • Thompson, Natalie J.
    • Affiliation: College of Arts and Sciences, Department of Chemistry
Abstract
  • The study of protein structure is crucial to the understanding of protein function as the two are inter dependent. Protein structure analysis is especially important as misfolded proteins often result in fatal diseases. Gas phase protein structure analysis provides a way to study the forces the effects of solvent and involved in the determination of protein structure. Mass spectrometry (MS) is a rapid and sensitive technique that can be used for the analysis of gas phase protein structure. Electron capture dissociation (ECD) tandem mass spectrometry (MS/MS) has been used in the analysis of primary, secondary, and tertiary gas phase protein structure. ECD MS/MS has proved to be exceptionally applicable in the study of protein structure due to the ability to cleave protein backbone bonds with the retention of noncovalent interactions and small molecule post translational modifications. The work described in this dissertation expands the use ECD and other electron capture techniques for protein structure analysis. All of the experiments were performed using a unique and versatile hybrid linear ion trap / time of flight (LIT/TOF) mass spectrometer capable of multiple ion activation techniques, such as collision induced dissociation (CID), infrared multiphoton dissociation (IRMPD), ECD, hot ECD (HECD), and activated ion ECD (AI ECD). IR activation in conjunction with ECD (AI ECD) to disrupt noncovalent interactions was used to enhance primary structure analysis and monitor changes in tertiary structure via gas phase protein unfolding. The structure of the z type product ions was probed using ion molecule reactions, and the formation different structures was found to depend on the electron capture technique used to dissociate the parent ion. The work described in this dissertation demonstrates the use of AI ECD for analysis of gas phase protein structure. Also, the first analysis of z ion structure from various electron capture techniques is presented. This work highlights the versatility and utility of the LIT TOF for ECD MS/MS as an alternative to the commercial Fourier transform ion cyclotron resonance (FTICR) mass spectrometers.
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  • In Copyright
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  • "... in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Chemistry."
Advisor
  • Glish, Gary
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  • Chapel Hill, NC
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  • Open access
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