The objective of this research was to investigate the structure and catalytic mechanisms of the FosX protein class. These enzymes are epoxide hydrolases that disrupt the first step of bacterial cell wall biosynthesis, thereby inactivating the antibiotic fosfomycin. Data presented in this work concerns FosX enzymes from the genome of pathogenic microorganism Listeria monocytogenes and a recently discovered integron isolated from Pseudomonas putida. Hydrogen-deuterium exchange mass spectrometry was used to determine structural perturbations that occur when the Listeria protein binds to divalent metal species. In addition, steady-state metal binding to Listeria FosX was investigated using optical spectroscopy. Finally, the putative FosX gene from Pseudomonas was cloned and the protein product expressed and purified. The enzyme was tested for FosX activity and ability to confer fosfomycin resistance. The ultimate goal of this research is to elucidate further mechanistic details about this class of fosfomycin resistance proteins, with the hope to one day develop inhibitors that will lessen or eliminate bacterial resistance to this antibiotic.
| Identifer | oai:union.ndltd.org:VANDERBILT/oai:VANDERBILTETD:etd-11212006-082620 |
| Date | 01 December 2006 |
| Creators | Grandillo, Jessica Louise |
| Contributors | Dr. Richard Armstrong, Dr. Daniel Liebler |
| Publisher | VANDERBILT |
| Source Sets | Vanderbilt University Theses |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.vanderbilt.edu/available/etd-11212006-082620/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to Vanderbilt University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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