dc.contributor.advisor | Steven R. Tannenbaum. | en_US |
dc.contributor.author | Kim, Ji-Eun, 1974- | en_US |
dc.contributor.other | Massachusetts Institute of Technology. Biological Engineering Division. | en_US |
dc.date.accessioned | 2005-09-27T18:46:07Z | |
dc.date.available | 2005-09-27T18:46:07Z | |
dc.date.copyright | 2004 | en_US |
dc.date.issued | 2004 | en_US |
dc.identifier.uri | http://hdl.handle.net/1721.1/28865 | |
dc.description | Thesis (Ph. D.)--Massachusetts Institute of Technology, Biological Engineering Division, 2004. | en_US |
dc.description | Includes bibliographical references. | en_US |
dc.description.abstract | (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 | en_US |
dc.description.abstract | 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 | en_US |
dc.description.statementofresponsibility | by Ji-Eun Kim. | en_US |
dc.format.extent | 165 p. | en_US |
dc.format.extent | 8638393 bytes | |
dc.format.extent | 8659094 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | application/pdf | |
dc.language.iso | en_US | |
dc.publisher | Massachusetts Institute of Technology | en_US |
dc.rights | 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. | en_US |
dc.rights.uri | http://dspace.mit.edu/handle/1721.1/7582 | |
dc.subject | Biological Engineering Division. | en_US |
dc.title | Regulation of tumor necrosis factor-alpha induced apoptosis via posttranslational modifications in a human colon adenocarcinoma cell line | en_US |
dc.type | Thesis | en_US |
dc.description.degree | Ph.D. | en_US |
dc.contributor.department | Massachusetts Institute of Technology. Department of Biological Engineering | |
dc.identifier.oclc | 60412373 | en_US |