Obtaining Structural Insights on Bacterial Protein Complexes Using Time-Resolved Hydrogen-Deuterium Exchange Mass Spectrometry
dc.contributor.advisor | Audette, Gerald F. | |
dc.creator | Lento, Cristina | |
dc.date.accessioned | 2015-12-16T19:30:30Z | |
dc.date.available | 2015-12-16T19:30:30Z | |
dc.date.copyright | 2015-08-25 | |
dc.date.issued | 2015-12-16 | |
dc.date.updated | 2015-12-16T19:30:30Z | |
dc.degree.discipline | Chemistry | |
dc.degree.level | Master's | |
dc.degree.name | MSc - Master of Science | |
dc.description.abstract | Persistent infections by Pseudomonas aeruginosa are initiated by interaction of a type IV pilus (T4P) with receptors on the mucosal cells of susceptible hosts. Here, we examine the structural changes occurring between the monomeric and dimeric states of ΔK122 using time-resolved electrospray ionization hydrogen-deuterium exchange mass spectrometry (TRESI-HDX-MS). Based on levels of deuterium uptake, the N-terminal α-helix and the loop connecting the second and third strands of the anti-parallel β-sheet contribute significantly to pilin dimerization. Conversely, the antiparallel β-sheet and αβ loop region exhibit increased flexibility, while the receptor binding domain retains a rigid conformation in the equilibrium state. Additionally, Escherichia coli are able to adapt to changing environmental conditions and develop antibiotic resistance through a process called F-plasmid conjugation, carried out through a type IV secretion system (T4SS). The F-T4SS protein TraF is of particular interest due to its involvement in pilus assembly to mediate the transfer of DNA. Dynamic analysis of a GST-TraF construct through TRESI-HDX-MS was performed to gain further insights on its structure. These studies have revealed that the C-terminal region predicted to contain the thioredoxin-like domain is quite structured compared to the more solvent accessible N-terminal region predicted to form a protein-protein interaction with companion T4SS protein TraH. Structural analysis of a GST-TraF construct is on-going to further characterize the regions responsible for protein-protein interaction and the elucidation of its three-dimensional structure. | |
dc.identifier.uri | http://hdl.handle.net/10315/30744 | |
dc.language.iso | en | |
dc.rights | Author owns copyright, except where explicitly noted. Please contact the author directly with licensing requests. | |
dc.subject | Chemistry | |
dc.subject | Biochemistry | |
dc.subject | Biology | |
dc.subject.keywords | chemistry | |
dc.subject.keywords | mass spectrometry | |
dc.subject.keywords | biochemistry | |
dc.subject.keywords | protein dynamics | |
dc.subject.keywords | type IV pilins | |
dc.subject.keywords | protein nanotubes | |
dc.subject.keywords | bacterial conjugation | |
dc.subject.keywords | time-resolved electrospray ionization | |
dc.subject.keywords | hydrogen-deuterium exchange | |
dc.title | Obtaining Structural Insights on Bacterial Protein Complexes Using Time-Resolved Hydrogen-Deuterium Exchange Mass Spectrometry | |
dc.type | Electronic Thesis or Dissertation |
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