رسالة جامعية
Systemic exploration of redox proteome in human A431 cells under different Photofrin-mediated photodynamic treatment conditions by quantitative proteomics approach
| العنوان: | Systemic exploration of redox proteome in human A431 cells under different Photofrin-mediated photodynamic treatment conditions by quantitative proteomics approach |
|---|---|
| العنوان البديل: | 利用定量蛋白質體學技術平台系統性分析人類A431細胞在不同光動力療法條件下之氧化還原蛋白質體 |
| المؤلفون: | Yi Fang Yang, 楊宜芳 |
| مرشدي الرسالة: | J. S. Yu, 余兆松 |
| سنة النشر: | 2012 |
| المجموعة: | National Digital Library of Theses and Dissertations in Taiwan |
| الوصف: | 100 Photodynamic therapy (PDT) is a minimally invasive therapeutic clinical treatment for some cancer and non-tumor disorders. This process requires a photosensitizer which can absorb light and produce reactive oxygen species to damage biomolecules in target cells. Photofrin is the most widely used photosensitizer. Our previous studies showed that different distribution of Photofrin in cells results in distinct cell death after PDT and Photofrin can selectively interact with some proteins. Using human epidermoid carcinoma A431 cells as model, we herein applied SILAC-based quantitative proteome approach to systemically analyze the relationship between the subcellular location of Photofrin and the Photofrin-PDT-mediated protein oxidization (on Met residues). In addition, we try to identify Photofrin-interacting proteins in A431 cells. Our data showed that when Photofrin mainly localized to plasma membrane (condition I), intracellular organelles (condition II) and whole cell (condition III), there were 116, 84 and 199 proteins quantified greater than mean+2SD Met-oxidized proteins, respectively. Further analysis revealed that (i) the percentage of highly oxidized membrane proteins is significantly higher in condition I (13.33%) than in condition II (3.03%) and III (0.76%); (ii) the percentage of highly oxidized Golgi proteins is drastically higher in condition II (18.18%) than in condition I (3.33%) and III (9.09%); and (iii) the percentage of highly oxidized mitochondrial proteins is largely higher in condition III (30.3%) than in condition I (4.44%) and II (15.66%). The results indicate that PDT with Photofrin targeted to distinct subcellular localizations can affect the redox proteome in a site-specific manner in living cells. Regarding the Photofrin-interacting proteins, we have identified 66 such candidates via in vitro pull down assay (using Photofrin-coupled beads) combined with MS analysis. Among those highly oxidized proteins detected post Photofrin-PDT, we select protein A and B for further study. Mapping the observed oxidized Met residues into the 3D structure of the two proteins showed that these oxidized Met residues mainly distribute on the surface of the protein. We also provided evidence to show that Photofrin-PDT can directly inhibit the activity of protein A in vitro. Collectively, our data demonstrate for the first time that the relationship between protein Met oxidation and PDT with site-specific location of Photofrin in living cells. Our results also unravel many potential Photofrin-interacting proteins that deserve further investigation. |
| Original Identifier: | 100CGU05114078 |
| نوع الوثيقة: | 學位論文 ; thesis |
| وصف الملف: | 161 |
| الإتاحة: | http://ndltd.ncl.edu.tw/handle/52121421771566331555Test |
| رقم الانضمام: | edsndl.TW.100CGU05114078 |
| قاعدة البيانات: | Networked Digital Library of Theses & Dissertations |
| الوصف غير متاح. |