{"id":{"repo_id":"buffalo","oai_identifier":"oai:ubir.buffalo.edu:10477/86430"},"canonical_url":"https://search.dev.ndltd.org/etd/buffalo/oai:ubir.buffalo.edu:10477/86430","repository":{"repo_id":"buffalo","name":"Buffalo","base_url":"https://ubir.buffalo.edu/oai/request"},"display":{"title":"Structure, Function, and Inhibition of Aerobactin Biosynthesis from Hypervirulent Klebsiella pneumoniae","abstract":"Ph.D.","abstract_html":"Ph.D.","abstract_has_math":false,"creators":["Bailey, Daniel; 0000-0003-4126-8700"],"institution":"State University of New York at Buffalo","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Gulick, Andrew","Structural Biology"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025-02-21T17:22:20Z","date_published":"2025-02-21T17:22:20Z","updated_at":"2026-07-27T19:05:32Z","subjects":["biochemistry","biology","biophysics"],"languages":["eng"],"rights":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/10477/86430","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Gulick, Andrew","Structural Biology"]},{"key":"dc:creator","label":"Author","values":["Bailey, Daniel; 0000-0003-4126-8700"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2025-02-21T17:22:20Z","2020"]},{"key":"dc:publisher","label":"Institution","values":["State University of New York at Buffalo"]},{"key":"dc:type","label":"Dc Type","values":["Text","Dissertation"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["biochemistry","biology","biophysics"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/10477/86430"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Ph.D.","Since it was initially described in the mid-1980s in the Asian Pacific Rim, a hypervirulent pathotype of Klebsiella pneumoniae (hvKP) has disseminated throughout the globe. In contrast to classical opportunistic strains of Klebsiella pneumoniae, hvKP is able to cause serious life-threatening infections in previously healthy individuals in the community. Recent reports have confirmed fears within the medical community that the convergence of multi-drug resistant and hypervirulent KP pathotypes has led to the evolution of a highly transmissible, drug resistant, and virulent \"super bug.\" Contemporary investigations toward understanding the enhanced virulence of hvKP strains have highlighted the importance of the biosynthesis of the siderophore aerobactin. Siderophores are small molecule iron-chelators that allow bacteria to assimilate sufficient quantities of this vital nutrient in the often severely iron-limited host environment. With an ever-increasing demand for novel therapeutic approaches for treating Gram-negative infections, we hypothesized aerobactin could be a viable \"antivirulence\" target for the treatment of infections with hvKP and other pathogens that rely on this siderophore. The research presented herein focused on three general topics: (1) demonstrating the function of the enzymes required to biosynthesize aerobactin, (2) characterizing the structure of the aerobactin synthetase enzymes IucA and IucC, and (3) the developing a high-throughput screening platform for identifying novel small-molecule inhibitors of aerobactin biosynthesis. To lay the foundation, the aerobactin biosynthetic pathway was first functionally demonstrated in vitro using purified enzymes. X-ray crystallography and solution scattering analyses were employed to structurally characterize IucA and IucC, which were combined with a number of biochemical studies to propose a molecular catalytic mechanism for these enzymes. Finally, leveraging our functional and structural knowledge of IucA, a high-throughput biochemical assay was developed and employed to screen over 110,00 compounds for antagonism of IucA catalysis; identifying a number of inhibitors with in vitro activity at low-micromolar concentrations.","**To request an accessible version of the file(s) associated with this item, contact library@buffalo.edu. Please include the item's persistent URL [http://hdl.handle.net/. . .] in your request.**"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Structure, Function, and Inhibition of Aerobactin Biosynthesis from Hypervirulent Klebsiella pneumoniae"]}]}],"canonical_facts":{"dc:contributor":["Gulick, Andrew","Structural Biology"],"dc:creator":["Bailey, Daniel; 0000-0003-4126-8700"],"dc:date":["2025-02-21T17:22:20Z","2020"],"dc:description":["Ph.D.","Since it was initially described in the mid-1980s in the Asian Pacific Rim, a hypervirulent pathotype of Klebsiella pneumoniae (hvKP) has disseminated throughout the globe. In contrast to classical opportunistic strains of Klebsiella pneumoniae, hvKP is able to cause serious life-threatening infections in previously healthy individuals in the community. Recent reports have confirmed fears within the medical community that the convergence of multi-drug resistant and hypervirulent KP pathotypes has led to the evolution of a highly transmissible, drug resistant, and virulent \"super bug.\" Contemporary investigations toward understanding the enhanced virulence of hvKP strains have highlighted the importance of the biosynthesis of the siderophore aerobactin. Siderophores are small molecule iron-chelators that allow bacteria to assimilate sufficient quantities of this vital nutrient in the often severely iron-limited host environment. With an ever-increasing demand for novel therapeutic approaches for treating Gram-negative infections, we hypothesized aerobactin could be a viable \"antivirulence\" target for the treatment of infections with hvKP and other pathogens that rely on this siderophore. The research presented herein focused on three general topics: (1) demonstrating the function of the enzymes required to biosynthesize aerobactin, (2) characterizing the structure of the aerobactin synthetase enzymes IucA and IucC, and (3) the developing a high-throughput screening platform for identifying novel small-molecule inhibitors of aerobactin biosynthesis. To lay the foundation, the aerobactin biosynthetic pathway was first functionally demonstrated in vitro using purified enzymes. X-ray crystallography and solution scattering analyses were employed to structurally characterize IucA and IucC, which were combined with a number of biochemical studies to propose a molecular catalytic mechanism for these enzymes. Finally, leveraging our functional and structural knowledge of IucA, a high-throughput biochemical assay was developed and employed to screen over 110,00 compounds for antagonism of IucA catalysis; identifying a number of inhibitors with in vitro activity at low-micromolar concentrations.","**To request an accessible version of the file(s) associated with this item, contact library@buffalo.edu. Please include the item's persistent URL [http://hdl.handle.net/. . .] in your request.**"],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/10477/86430"],"dc:language":["eng"],"dc:publisher":["State University of New York at Buffalo"],"dc:rights":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."],"dc:subject":["biochemistry","biology","biophysics"],"dc:title":["Structure, Function, and Inhibition of Aerobactin Biosynthesis from Hypervirulent Klebsiella pneumoniae"],"dc:type":["Text","Dissertation"]},"updated_at":"2026-07-27T19:05:32Z"}