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研究生: 王于彰
Wang, Yu-Zhang
論文名稱: Smad4蛋白之表現與分子對接之研究
Expression of Smad4 protein and its molecular docking research
指導教授: 蕭世裕
Shaw, Shyh-Yu
學位類別: 碩士
Master
系所名稱: 理學院 - 化學系
Department of Chemistry
論文出版年: 2024
畢業學年度: 112
語文別: 中文
論文頁數: 49
中文關鍵詞: Smad4蛋白基質金屬蛋白酶Smad pathway轉化生長因子-β
外文關鍵詞: Matrix metalloproteinase-9, Smad pathway, Smad4, TGF-β
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  • 中文摘要 Ⅰ 英文摘要 Ⅱ 致謝 Ⅸ 一、 緒論 1 1. 口腔鱗狀細胞癌 1 2. 轉化生長因子-β 1 2.1 Smad dependent pathway 2 2.2 Smad independent pathway 2 3. 基質金屬蛋白酶 3 3.1 MMP簡介與功能 3 3.2 MMP歷史與分類 3 3.3 MMP在腫瘤生長中的作用 4 4. Smad蛋白家族 5 4.1調控型(R-Smad) 5 4.2通用型(Co-Smad) 5 4.3抑制型(I-Smad) 5 4.4 Smad4蛋白結構及功能介紹 6 5. 四環黴素 7 5.1四環黴素抗生素特性 7 5.2四環黴素非抗生素特性 7 6. 蛋白質結晶 8 7. 分子對接 10 二、研究動機 11 三、藥品耗材 12 藥品 12 儀器 13 四、實驗步驟 14 1. Transform至E.coli DH5α 14 2. 萃取質體DNA 14 3. 以限制酵素切割pGEX-4T1-smad4質體 15 4. 聚合酶連鎖反應Colony PCR 15 5. Transform至E.coli BL21(DE3) 16 6. 養菌 16 7. 破菌 17 8. 純化 17 9. Smad4蛋白之十二烷基硫酸鈉聚丙烯醯胺凝膠電泳分析 19 10. 分子對接 20 五、結果與討論 21 1. 以Colony PCR確認Smad4 DNA 21 2. pGEX4T1-Smad4質體切割 21 3. Smad4蛋白之表現及SDS-PAGE分析 22 4. Smad4之蛋白質養晶 23 5. 分子對接之結果 23 六、結論25 七、參考文獻26 附錄-圖例 30 圖一、pGEX4T1-Smad4 colony PCR結果 30 圖二、以限制酵素切割pGEX4T1-Smad4質體 30 圖三、Smad4 SDS-PAGE分析結果 31 圖四、以Nanodrop所測得之Smad4濃度 32 圖五、Smad4與強力黴素間之鍵結情形 32 圖六、Smad4與強力黴素分子模擬結果 33 附錄-表格 34 表一、Smad4基因擴增PCR cycle 34 表二、Smad4 primer序列 34 附錄一、pGEX4T1質體圖 35

    林志強, 四環黴素抑制口腔鱗狀癌細胞內基質金屬蛋白酶-9表現量之抑制機制的研究,碩士論文。國立成功大學生物科技研究所碩士班。2013。

    陳宥維。Smad4 MH2蛋白質晶體結構解析度優化及與四環黴素類似物之分子對接研究,碩士論文。國立成功大學化學系碩士班。2023。

    張文馨。基因重組人類Smad4蛋白之表現與功能分析,碩士論文。國立成功大學化學系碩士班。2015。

    劉蒼維,et al. 骨形態發生蛋白信號通路在牙根發育中的作用. 華西口腔醫學雜誌, 2018,36(5);559-563.

    Kohei Miyazono, transforming growth factor-b signaling in epithelial-mesenchymal transition and progression of cancer. Proceedings of the Japan Academy,Series B,2009.85

    Bernhard Schmierer & Caroline S. Hill, TGFβ–SMAD signal transduction: molecular specificity and functional flexibility, Nature Reviews Molecular Cell Biology volume 8, pages970–982 (2007)

    Jeff Yat-Fai Chung,et al, TGF-β Signaling: From Tissue Fibrosis to Tumor Microenvironment, Int. J. Mol. Sci. 2021, 22(14), 7575;
    Kalliopi Tzavlaki&Aristidis Moustakas, TGF-β Signaling, Biomolecules. 2020 Mar; 10(3): 487.
    Niina Reunanen and VeliMatti Kähäri., Matrix Metalloproteinases in Cancer Cell Invasion, Landes Bioscience; 2000-2013.
    Aki Hanyu,Yasuhiro Ishidou,et al, The N domain of Smad7 is essential for specific inhibition of transforming growth factor-β signaling, J Cell Biol (2001) 155 (6): 1017–1028.

    Maria J. Macias,et al, Structural determinants of Smad function in TGF-β signaling,Trend in biochemical science, June 2015, Pages 296-308

    Bin Qin,et al, Crystal structure of a transcriptionally active Smad4 fragment,Structure, January 1999, Pages 1493-1503
    Ying Gu,et al, Non-antibacterial tetracycline formulations: clinical applications in dentistry and medicine,Journal of Microbiology, 4(1)
    Trudy H. Grossman, Tetracycline Antibiotics and Resistance,Cold Spring Harbor Perspectives in Medicine, March 17, 2016
    N.S. Ramamurthy,et al, The effect of alloxan diabetes on gingival collagen metabolism in rats, Archives of Oral Biology, November 1972, Pages 1551-1560
    McPherson, A. & Gavira, J. A. (2014). Introduction to protein crystallization. Acta Cryst.F70,2-20

    Cabral-Pacheco GA, Garza-Veloz I, Castruita-De la Rosa C, Ramirez-Acuña JM, Perez-Romero BA, Guerrero-Rodriguez JF, Martinez-Avila N, Martinez-Fierro ML. The Roles of Matrix Metalloproteinases and Their Inhibitors in Human Diseases. Int J Mol Sci. 2020 Dec 20;21(24):9739.

    Page-McCaw, A., Ewald, A. & Werb, Z. Matrix metalloproteinases and the regulation of tissue remodelling. Nat Rev Mol Cell Biol 8, 221–233 (2007).

    Laronha H, Caldeira J. Structure and Function of Human Matrix Metalloproteinases. Cells. 2020 Apr 26;9(5):1076.

    GROSS J, LAPIERE CM. Collagenolytic activity in amphibian tissues: a tissue culture assay. Proc Natl Acad Sci U S A. 1962 Jun 15;48(6):1014-22.

    Van Lint P, Libert C. Chemokine and cytokine processing by matrix metalloproteinases and its effect on leukocyte migration and inflammation. . Biol.December 2007, 82 (6): 1375–81

    T. Vartio, A. Vaheri. 1981. A gelatin-binding 70,000-dalton glycoprotein synthesized distinctly from fibronectin by normal and malignant adherent cells. J. Biol. Chem. 256: 13085–13090.

    M.S. Hibbs, K.A. Hasty, J.M. Seyer, A.H. Kang, et al. 1985. Biochemical and immunological characterization of the secreted forms of human neutrophil gelatinase. J. Biol. Chem. 260: 2493–2500.

    L.P. Kotra, L. Zhang, R. Fridman, R. Orlando, et al. 2002. Glycosylation pattern of the zymogenic form of human matrix metalloproteinase-9. Bioorg. Chem. 30: 356–370.

    Robinson PN, Arteaga-Solis E, Baldock C, et al The molecular genetics of Marfan syndrome and related disorders Journal of Medical Genetics 2006;43:769-787.
    Jennifer P. Habashi et al.Losartan, an AT1 Antagonist, Prevents Aortic Aneurysm in a Mouse Model of Marfan Syndrome.Science312,117-121(2006).
    Blobe GC, Schiemann WP, Lodish HF. Role of transforming growth factor beta in human disease. N. Engl. J. Med. May 2000, 342 (18): 1350–8
    Massague J. Receptors for the TGF-beta family. Cell. 1992 Jun 26;69(7):1067-70

    Shi J, Yoshino O, Osuga Y, Nishii O, Yano T, Taketani Y (March 2010). "Bone morphogenetic protein 7 (BMP-7) increases the expression of follicle-stimulating hormone (FSH) receptor in human granulosa cells"
    De Caestecker, Mark P. (2000). "The Smad4 Activation Domain (SAD) Is a Proline-rich, p300-dependent Transcriptional Activation Domain". The Journal of Biological Chemistry. 275 (3): 2115–2122.31.
    Baburajendran, Nithya; Jauch, Ralf; Tan, Clara Yueh Zhen; Narasimhan, Kamesh; Kolatkar, Prasanna R. (2011). "Structural basis for the cooperative DNA recognition by Smad4 MH1 dimers". Nucleic Acids Research. 39 (18): 8213–8222.
    Kumar R, Kumar R, Tanwar P. Structural based screening of potential inhibitors of SMAD4: a step towards personalized medicine for gall bladder and other associated cancers. Mol Divers. 2021 Aug;25(3):1945-1961.

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