Regulation of tumor necrosis factor-alpha induced apoptosis via posttranslational modifications in a human colon adenocarcinoma cell line
Thesis (Ph. D.)--Massachusetts Institute of Technology, Biological Engineering Division, 2004.
| Main Author: | |
|---|---|
| Other Authors: | |
| Format: | Thesis |
| Language: | en_US |
| Published: |
Massachusetts Institute of Technology
2005
|
| Subjects: | |
| Online Access: | http://hdl.handle.net/1721.1/28865 |
| _version_ | 1811073340359573504 |
|---|---|
| author | Kim, Ji-Eun, 1974- |
| author2 | Steven R. Tannenbaum. |
| author_facet | Steven R. Tannenbaum. Kim, Ji-Eun, 1974- |
| author_sort | Kim, Ji-Eun, 1974- |
| collection | MIT |
| description | Thesis (Ph. D.)--Massachusetts Institute of Technology, Biological Engineering Division, 2004. |
| first_indexed | 2024-09-23T09:31:32Z |
| format | Thesis |
| id | mit-1721.1/28865 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:31:32Z |
| publishDate | 2005 |
| publisher | Massachusetts Institute of Technology |
| record_format | dspace |
| spelling | mit-1721.1/288652019-04-12T14:35:33Z Regulation of tumor necrosis factor-alpha induced apoptosis via posttranslational modifications in a human colon adenocarcinoma cell line Kim, Ji-Eun, 1974- Steven R. Tannenbaum. Massachusetts Institute of Technology. Biological Engineering Division. Massachusetts Institute of Technology. Biological Engineering Division. Biological Engineering Division. Thesis (Ph. D.)--Massachusetts Institute of Technology, Biological Engineering Division, 2004. Includes bibliographical references. (cont.) phosphoproteomics technology, IMAC/LC/MS/MS, [approximately] 200 phosphosites were identified from HT-29 cells, some of which were detected only from insulin-treated cells. Our phosphoproteomics approach also enabled us to detect alteration of both known and unknown phosphorylation states of apoptosis-related proteins at two time points during early apoptosis induced by tumor necrosis factor-α Apoptosis, a physiologically regulated cell death, plays critical roles in development and immune system by maintaining tissue homeostasis. The thesis project investigates regulations of apoptosis in a human colon adenocarcinoma cell line, HT-29, exposed to diverse cellular stimuli, focusing on a specific protein as well as global level of proteins. The first part of the thesis demonstrated S-nitrosation of procaspase-9. S-nitrosation is a novel protein modification to regulate protein-protein interaction or protein activity. This modification has been implied to inactivate caspases. We could visualize S-nitrosation of an initiator caspase, procaspase-9, by enriching low-abundant procaspase-9 with immunoprecipitation and stabilizing S-nitroso-cysteine with biotin labeling. Nitric oxide synthase inhibitors and tumor necrosis factor-α (TNF-α) reduced the S-nitrosation level of procaspase-9, suggesting that S-nitrosation may be regulated by a nitric oxide synthase and denitrosation is likely a mechanism of apoptosis. The second part of the thesis is to examine survival effects of insulin on cells undergoing TNF-α-induced apoptosis. Insulin decreased the TNF-α-induced cleavage of key apoptotic mediators, caspases, and their substrates as well as apoptosis, in part, depending on phosphatidylinositol-3 kinase (PI-3K)/Akt pathway. One of protective mechanisms by insulin is likely to decrease the TNF-α-induced dissociation of a potent inhibitor of caspases, X-chromosome linked inhibitor of apoptosis protein (XIAP), from procaspase-9 via PI-3K/Akt pathway. Lack of phosphoproteomics data in HT-29 cells led the third part of the thesis to focus on investigating global level regulation of phosphoproteins during apoptosis. With a by Ji-Eun Kim. Ph.D. 2005-09-27T18:46:07Z 2005-09-27T18:46:07Z 2004 2004 Thesis http://hdl.handle.net/1721.1/28865 60412373 en_US MIT theses are protected by copyright. They may be viewed, downloaded, or printed from this source but further reproduction or distribution in any format is prohibited without written permission. http://dspace.mit.edu/handle/1721.1/7582 165 p. 8638393 bytes 8659094 bytes application/pdf application/pdf application/pdf Massachusetts Institute of Technology |
| spellingShingle | Biological Engineering Division. Kim, Ji-Eun, 1974- Regulation of tumor necrosis factor-alpha induced apoptosis via posttranslational modifications in a human colon adenocarcinoma cell line |
| title | Regulation of tumor necrosis factor-alpha induced apoptosis via posttranslational modifications in a human colon adenocarcinoma cell line |
| title_full | Regulation of tumor necrosis factor-alpha induced apoptosis via posttranslational modifications in a human colon adenocarcinoma cell line |
| title_fullStr | Regulation of tumor necrosis factor-alpha induced apoptosis via posttranslational modifications in a human colon adenocarcinoma cell line |
| title_full_unstemmed | Regulation of tumor necrosis factor-alpha induced apoptosis via posttranslational modifications in a human colon adenocarcinoma cell line |
| title_short | Regulation of tumor necrosis factor-alpha induced apoptosis via posttranslational modifications in a human colon adenocarcinoma cell line |
| title_sort | regulation of tumor necrosis factor alpha induced apoptosis via posttranslational modifications in a human colon adenocarcinoma cell line |
| topic | Biological Engineering Division. |
| url | http://hdl.handle.net/1721.1/28865 |
| work_keys_str_mv | AT kimjieun1974 regulationoftumornecrosisfactoralphainducedapoptosisviaposttranslationalmodificationsinahumancolonadenocarcinomacellline |