Dengue virus (DENV) is a mosquito-transmitted flavivirus that threatens approximately half of the world's population. In this dissertation, the use of bioorthogonal chemistry as a tool for researching emerging viral diseases, including DENV is explored. To this end, a bioorthogonally-modified amino acid was successfully installed within the proteome of DENV, which was used for the pull down of a known virus-protein interaction. This technology is intended to be broadly used for the determination of any virus-host interaction, through the installment of a non-perturbing modification that 1) does not hinder viral infectivity and 2) can be selectively discriminated by any complimentary probe. En route to using this technology, a new viral purification strategy was developed for DENV that reduces the overall purification time by 10 hours, and improves retention of virion infectivity. This method and a survey of other viral purification methods used with DENV is contained herein. Furthermore, a chemical scaffold that was repurposed for exploration of protein-protein crosslinking, namely for release of a reactive chemical warhead under acidic conditions, was used for the surface modification of DENV. This triazabutadiene probe was found to be activated by light. In this dissertation is reported the first time aryl diazonium ions for protein crosslinking have been generated on a protein or viral surface through UV-irradiation. The advantages and limitations of this chemistry are presented herein.
| Identifer | oai:union.ndltd.org:arizona.edu/oai:arizona.openrepository.com:10150/621769 |
| Date | January 2016 |
| Creators | Jensen, Stephanie Meryl, Jensen, Stephanie Meryl |
| Contributors | Jewett, John C., Jewett, John, Ghosh, Indraneel, Marty, Michael, Montfort, William |
| Publisher | The University of Arizona. |
| Source Sets | University of Arizona |
| Language | en_US |
| Detected Language | English |
| Type | text, Electronic Dissertation |
| Rights | Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author. |
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