Understanding collagen-l folding and misfolding
Thesis: S.B. in Chemistry and Biology, Massachusetts Institute of Technology, Department of Chemistry, 2015.
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| Formato: | Tese |
| Idioma: | eng |
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Massachusetts Institute of Technology
2015
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| Acesso em linha: | http://hdl.handle.net/1721.1/98783 |
| _version_ | 1826202824104476672 |
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| author | Taylor, Rebecca J. S.B. Massachusetts Institute of Technology |
| author2 | Matthew D. Shoulders. |
| author_facet | Matthew D. Shoulders. Taylor, Rebecca J. S.B. Massachusetts Institute of Technology |
| author_sort | Taylor, Rebecca J. S.B. Massachusetts Institute of Technology |
| collection | MIT |
| description | Thesis: S.B. in Chemistry and Biology, Massachusetts Institute of Technology, Department of Chemistry, 2015. |
| first_indexed | 2024-09-23T12:20:20Z |
| format | Thesis |
| id | mit-1721.1/98783 |
| institution | Massachusetts Institute of Technology |
| language | eng |
| last_indexed | 2024-09-23T12:20:20Z |
| publishDate | 2015 |
| publisher | Massachusetts Institute of Technology |
| record_format | dspace |
| spelling | mit-1721.1/987832019-04-12T07:53:23Z Understanding collagen-l folding and misfolding Taylor, Rebecca J. S.B. Massachusetts Institute of Technology Matthew D. Shoulders. Massachusetts Institute of Technology. Department of Chemistry. Massachusetts Institute of Technology. Department of Chemistry. Chemistry. Thesis: S.B. in Chemistry and Biology, Massachusetts Institute of Technology, Department of Chemistry, 2015. Cataloged from PDF version of thesis. Includes bibliographical references (pages 74-79). Chapter One: Introduction to Type I Collagen and Osteogenesis Imperfecta Collagen-I is the primary proteinaceous component of skin, bone, and tendon. Disruptions in collagen-I homeostasis, typically due to non-synonymous mutations in collagen-- encoding genes, cause a variety of severe incurable diseases, including Osteogenesis Imperfecta (01). 01 phenotypes include brittle, deformed bones, frequent fractures, and growth deficiency. In order to fill the need for treatments that target the underlying causes of collagen-I-related diseases like 01, a better understanding of the collagen-I proteostasis network and how it differentially engages mutant and wild type collagen-1, is required. Chapter Two: Creation and Characterization of a Cell-Based Platform for Delineating the Wild Type and Mutant Collagen-I Proteostasis Network Previous studies of the collagen biosynthetic pathway have been limited by the lack of a biochemically tractable system to allow manipulation of the collagen-I genes (and other genes of interest) and especially by the lack of immunoprecipitation-grade antibodies for collagen-I which has prevented the broad study of the complete set of collagen-1 interacting proteins. We have overcome the challenges of working with the collagen-I genes and have created stable cell lines that inducibly express epitope-tagged versions of both wild type and mutant collagen-1. This platform is greatly facilitating studies of the collagen-I proteostasis network. Chapter Three: Mechanistic Exploration of Novel Collagen-I Interacting Proteins Identified by SILAC Mass Spectrometry Using the model cell platform described in Chapter 2, we have performed an unbiased and quantitative investigation into the network of collagen-I interacting proteins using SILACassisted, quantitative mass spectrometry. The method allowed us to identify more than 25 novel collagen-I interactors. We are currently investigating the mechanistic roles of these proteins in collagen-I processing using shRNA knockdown of proteins of interest. Chapter Four: Creation and Validation of Constructs for the Independent Expression of the Collagen-I C-Propeptide Domains The collagen-I C-propeptide domains are responsible for collagen-I chain selectivity and triple helix nucleation. Many unanswered questions remain relating to the mechanistic details of C-propeptide function both in collagen-I folding and also in diverse biological processes. In order to address these questions, we created and validated constructs that allow the independent expression of the C-propeptide domains. by Rebecca J. Taylor. S.B. in Chemistry and Biology 2015-09-17T19:11:45Z 2015-09-17T19:11:45Z 2015 2015 Thesis http://hdl.handle.net/1721.1/98783 921140945 eng M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission. http://dspace.mit.edu/handle/1721.1/7582 79 pages application/pdf Massachusetts Institute of Technology |
| spellingShingle | Chemistry. Taylor, Rebecca J. S.B. Massachusetts Institute of Technology Understanding collagen-l folding and misfolding |
| title | Understanding collagen-l folding and misfolding |
| title_full | Understanding collagen-l folding and misfolding |
| title_fullStr | Understanding collagen-l folding and misfolding |
| title_full_unstemmed | Understanding collagen-l folding and misfolding |
| title_short | Understanding collagen-l folding and misfolding |
| title_sort | understanding collagen l folding and misfolding |
| topic | Chemistry. |
| url | http://hdl.handle.net/1721.1/98783 |
| work_keys_str_mv | AT taylorrebeccajsbmassachusettsinstituteoftechnology understandingcollagenlfoldingandmisfolding |