Characterization of Structures and Dynamics of Intrinsically Disordered Domain and Proteins Using Time Resolved-Hydrogen Deuterium Exchange Mass Spectrometry

Simple item page
dc.contributor.advisorWilson, Derek J.
dc.creatorZhu, Shaolong
dc.date.accessioned2017-07-27T13:39:51Z
dc.date.available2017-07-27T13:39:51Z
dc.date.copyright2017-02-16
dc.date.issued2017-07-27
dc.date.updated2017-07-27T13:39:50Z
dc.degree.disciplineChemistry
dc.degree.levelDoctoral
dc.degree.namePhD - Doctor of Philosophy
dc.description.abstractProteins are inherently dynamic. Virtually all of the processes that underlie biological activity, including binding partner recognition, catalysis and allostery among many others, require transient adoption of specific high energy conformations. While many proteins contain highly dynamic regions or Intrinsically Disordered Domains (IDDs), Intrinsically Disordered Proteins (IDPs) are free of well-defined structural features altogether. These domains/proteins play integral roles in a wide variety of biological processes, but are often associated with neurodegenerative disease and cancer. Hence it is crucial to study these biomacromolecules to understand the structural underpinnings of their biological functions as well as the factors that drive disease pathology. However, many of the current high resolution structural techniques (like X-ray crystallography and NMR) are not feasible for such studies because of their reliance on the presence of a well-defined native conformation. An emerging structure labeling technique called Time Resolved-Hydrogen Deuterium Exchange (TR-HDX) can offer an alternative strategy. In this approach, structure-dependent hydrogen/deuterium labelling is measured on the millisecond timescale, providing an exquisitely sensitive picture of the weak hydrogen bonding networks that impart residual structure in IDDs and IDPs. In this work, TR-HDX was implemented onto microfluidic chip to achieve solution phase site-specific analysis of the structures and dynamics of p53 N-terminal transactivation domain and tau protein. Gas-phase TR-HDX was also implemented to study gas-phase protein structures that become populated during Differential Mobility Spectrometry (DMS) ion mobility analyses. Critical advances include the discovery and characterization of an amyloidogenic tau intermediate that may underlie Alzheimers disease pathology, a dynamics-based model for recruitment of co-factors to cancer protein p53 upon phosphorylation, a detailed account of how a set of anti-amyloid drug candidates affect tau structure and dynamics, and development of a method to predict solution-phase protein stability from DMS-HDX measurements.
dc.identifier.urihttp://hdl.handle.net/10315/33543
dc.language.isoen
dc.rightsAuthor owns copyright, except where explicitly noted. Please contact the author directly with licensing requests.
dc.subjectChemistry
dc.subject.keywordsIntrinsically disordered proteins
dc.subject.keywordsIntrinsically disordered domains
dc.subject.keywordsTau
dc.subject.keywordsp53
dc.subject.keywordsHyperphosphorylation
dc.subject.keywordsPhosphorylation
dc.subject.keywordsHydrogen deuterium exchange mass spectrometry
dc.subject.keywordsTime resolved hydrogen deuterium exchange
dc.subject.keywordsMass spectrometry
dc.subject.keywordsDifferential mobility spectrometry
dc.subject.keywordsIon mobility spectrometry
dc.subject.keywordsAlzheimer's disease
dc.subject.keywordsCancer
dc.subject.keywordsAmyloid
dc.subject.keywordsAnti-aggregation
dc.subject.keywordsGSK3beta
dc.subject.keywordsMicrofluidic chips
dc.subject.keywordschk2 kinase
dc.subject.keywordsPleckstrin homology domain
dc.subject.keywordsFPLC
dc.subject.keywordsLC
dc.subject.keywordsESI
dc.titleCharacterization of Structures and Dynamics of Intrinsically Disordered Domain and Proteins Using Time Resolved-Hydrogen Deuterium Exchange Mass Spectrometry
dc.typeElectronic Thesis or Dissertation

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Zhu_Shaolong_2017_PhD.pdf
Size:
4.34 MB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 2 of 2
No Thumbnail Available
Name:
license.txt
Size:
1.83 KB
Format:
Plain Text
Description:
Download
No Thumbnail Available
Name:
YorkU_ETDlicense.txt
Size:
3.38 KB
Format:
Plain Text
Description:
Download

Collections

Chemistry

All items in the YorkSpace institutional repository are protected by copyright, with all rights reserved except where explicitly noted.