Investigating the composition and architecture of insoluble polysaccharide systems and protein interactions
Abstract/Contents
- Abstract
- Polysaccharides are essential macromolecules found across all life forms. However, they can pose a challenge to solution-based analytical methods due to insolubility, high molecular weight, and close interactions with other biomolecules including proteins. To study these diverse polysaccharides and heterogeneous composites, we leverage solid-state nuclear magnetic resonance (ssNMR) to quantify and compare polysaccharide modifications, to determine the composition ratio of protein-polysaccharide mixtures, and to assign novel gene function. Additionally, we develop methods to enable cryogenic electron microscopy (cryo-EM) visualization of the architecture of these biologically complex systems. Here we report on studies of three systems: exopolysaccharides produced by S. meliloti, amyloid-polysaccharide biofilms formed by E. coli and S. Typhimurium, and chrysalides of the Monarch and Swallowtail butterfly. First, we demonstrate how two types of fibers in the bacterial extracellular matrix assemble into an impressive "basket-like" architecture and detail our specialized cryo-EM approaches optimized for the unique characteristics of the biofilm. After "unraveling" the biofilm baskets, we weave them back together by reconstituting robust biofilms with inter- and intra- species complementation between E. coli and S. Typhimurium. Next, we identify a novel pyruvyltransferase using ssNMR to analyze rhizobial exopolysaccharides. Finally, we perform a comparative analysis of a much larger yet comparable structure to the biofilm extracellular matrix - the butterfly chrysalis - detailing the chitin-rich framework and proteomic analysis of Monarch and Swallowtail chrysalides.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2022; ©2022 |
| Publication date | 2022; 2022 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Goularte, Nicolette Fatima |
|---|---|
| Degree supervisor | Cegelski, Lynette |
| Degree supervisor | Puglisi, Joseph D |
| Thesis advisor | Cegelski, Lynette |
| Thesis advisor | Puglisi, Joseph D |
| Thesis advisor | Chiu, Wah |
| Thesis advisor | Long, Sharon R |
| Degree committee member | Chiu, Wah |
| Degree committee member | Long, Sharon R |
| Associated with | Stanford University, Department of Structural Biology |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Nicolette F. Goularte. |
|---|---|
| Note | Submitted to the Department of Structural Biology. |
| Thesis | Thesis Ph.D. Stanford University 2022. |
| Location | https://purl.stanford.edu/yz548hy9417 |
Access conditions
- Copyright
- © 2022 by Nicolette Fatima Goularte
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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