Identification and characterization of inhibitors of amyloids and amyloid-based biofilm in E. coli through cell-based high-throughput screening
Abstract/Contents
- Abstract
- Biofilms are multicellular microbial communities that consist of the cells plus a self-produced extracellular matrix (ECM), a complex, heterogeneous housing that protects the cells from environmental insults. In the human host, biofilms can result in persistent infections that are difficult to treat, leading to further health complications and longer hospital stays. Additionally, environmental persistence through biofilm formation can contribute to foodborne outbreaks by pathogenic strains. Identification of compounds that inhibit biofilm formation would provide a valuable tool for reduction of pathogenesis resulting from biofilms. In natural biofilms, adhesive amyloid fibers are abundant and employed by a wide variety of bacteria, suggesting a crucial role in biofilm formation and persistence. The most well-studied bacterial amyloid adhesins are the curli fibers produced by Escherichia coli and Salmonella species. In the laboratory, the coproduction of curli and cellulose by E. coli enables the formation of bacterial biofilms on agar, characterized by the hallmark wrinkled colony morphology. Amyloid fibers are highly ordered protein aggregates formed by the non-covalent polymerization of a single protein with the polypeptide chains aligned in the cross-β-configuration. Originally considered to be exclusively the result of protein misfolding, many examples of amyloid fibers that perform physiologically useful and specialized functions for the organism producing them have been identified and termed functional amyloids. Understanding the mechanism of formation of these functional amyloids not only provides insight into the prevention of biofilm formation, but also the prevention of diseases associated with amyloids produced by protein mis-folding. We developed a cell-based high-throughput screen to identify inhibitors of curli-dependent biofilm formation by the uropathogenic E. coli clinical isolate UTI89, and, in parallel, curli-dependent cell adhesion by the laboratory strain MC4100. In screening the compound libraries available at Stanford's High Throughput Bioscience Center, several potential inhibitors were identified and subsequently characterized.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2016 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Maher, Marie Celeste |
|---|---|
| Associated with | Stanford University, Department of Chemistry. |
| Primary advisor | Cegelski, Lynette |
| Thesis advisor | Cegelski, Lynette |
| Thesis advisor | Boxer, Steven G. (Steven George), 1947- |
| Thesis advisor | Khosla, Chaitan, 1964- |
| Advisor | Boxer, Steven G. (Steven George), 1947- |
| Advisor | Khosla, Chaitan, 1964- |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Marie Celeste Maher. |
|---|---|
| Note | Submitted to the Department of Chemistry. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2016. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2016 by Marie Celeste Maher
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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