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dc.contributor.advisorWang, Rongsheng
dc.creatorLyu, Zhigang
dc.date.accessioned2021-08-23T18:18:18Z
dc.date.available2021-08-23T18:18:18Z
dc.date.issued2021
dc.identifier.urihttp://hdl.handle.net/20.500.12613/6892
dc.description.abstractPost-translational modifications (PTMs) diversify the functions and control the stability of proteins by the covalent addition of chemical groups or proteins. These modifications include phosphorylation, acetylation, glycosylation, methylation, ubiquitination, etc. and affect all aspects of cellular activities. Dysregulation of PTMs is often linked to aging, oncogenesis, and various autoimmune diseases. Proteins involved in the PTM writing, removing, and reading process are very important biomarkers and hot therapeutic targets. Yet, current chemical methodologies to globally profiling PTM substrates rely on coppercatalyzed azide-alkyne cycloaddition (CuAAC) reaction in which bulky alkyne or azide groups in length and size are used. The intrinsic steric hinderance limits the general application of chemical reporters to metabolic incorporation by PTM writers possessing spacious active pockets.This dissertation will start with the demonstration that the alpha fluorine to amide bond was a steric free chemical reporter in protein acetylation. Fluorinated cofactors or precursors could be utilized by acetyltransferases to modify peptides and proteins both in vitro and in live cells. A novel bioorthogonal reaction named as fluorine thiol displacement reaction (FTDR) using thiophenol as the warhead was developed for converting a fluorine label in biomolecules to a fluorophore or biotin tag. This whole platform showed great potential to label, image, and enrich acetylation substrates. Another goal of this dissertation was the evolution of FTDR to fluorine selenol displacement reaction (FSeDR). Aliphatic selenol, aromatic selenol and thiol derivatives were compared in parallel in the labelling of fluorinated small molecules and proteins. Aliphatic selenol displayed the best reactivity under the mild physiological condition and exhibited a high degree of chemical stability. The updated platform was then used in the SILAC-based quantitative proteomics study of acetylation in the prostate cancer cell PC-3. The final goal of this dissertation focuses on the study of steric-free bioorthogonal labeling of glycosylation substrates based on FSeDR. The fluorine-tag labelled unnatural monosaccharides could be utilized by glycotransferases in live cells to modify sialylated glycoproteins, N-glycoproteins, and O-GlcNAcylated proteins. With the help of FSeDR, the fluorine reporter was further functionalized to fluorescent probes or affinity tags for imaging or enrichment studies. The steric-free fluorine reporter has the potential to become a powerful chemical reporter for glycan labelling.
dc.format.extent252 pages
dc.language.isoeng
dc.publisherTemple University. Libraries
dc.relation.ispartofTheses and Dissertations
dc.rightsIN COPYRIGHT- This Rights Statement can be used for an Item that is in copyright. Using this statement implies that the organization making this Item available has determined that the Item is in copyright and either is the rights-holder, has obtained permission from the rights-holder(s) to make their Work(s) available, or makes the Item available under an exception or limitation to copyright (including Fair Use) that entitles it to make the Item available.
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/
dc.subjectChemistry
dc.subjectBiology
dc.subjectAcetylation
dc.subjectDisplacement reaction
dc.subjectFluorine
dc.subjectGlycosylation
dc.subjectProteomics
dc.subjectSteric-free
dc.titleSteric-Free Bioorthogonal Labeling of Post-translational Modification Substrates Based on A Fluorine-Thiol/Selenol Displacement Reaction
dc.typeText
dc.type.genreThesis/Dissertation
dc.contributor.committeememberSchafmeister, Christian
dc.contributor.committeememberNicholson, Allen W.
dc.contributor.committeememberBlass, Benjamin E.
dc.description.departmentChemistry
dc.relation.doihttp://dx.doi.org/10.34944/dspace/6874
dc.ada.noteFor Americans with Disabilities Act (ADA) accommodation, including help with reading this content, please contact scholarshare@temple.edu
dc.description.degreePh.D.
dc.identifier.proqst14633
dc.creator.orcid0000-0002-6903-3965
dc.date.updated2021-08-21T10:09:13Z
dc.embargo.lift08/17/2023
dc.identifier.filenamelyu_temple_0225E_14633.pdf


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