研究生: |
鄭堯聰 Yao-Tsung Cheng |
---|---|
論文名稱: |
化學蛋白質體學搭配質譜技術分析Sunitinib之標的候選蛋白質 Chemical Proteomics Analysis of Sunitinib Target Candidates by Mass Spectrometry |
指導教授: |
陳頌方
Chen, Sung-Fang |
學位類別: |
碩士 Master |
系所名稱: |
化學系 Department of Chemistry |
論文出版年: | 2013 |
畢業學年度: | 101 |
語文別: | 中文 |
論文頁數: | 113 |
中文關鍵詞: | 化學蛋白質體學 、Sunitinib 、質譜儀 、親和性層析 、急性骨髓性白血病 、活性為基礎的探針 |
英文關鍵詞: | Chemical proteomics, Sunitinib, Mass spectrometry, Affinity chromatography, Acute myeloid leukemia, Activity-based probe |
論文種類: | 學術論文 |
相關次數: | 點閱:108 下載:12 |
分享至: |
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報 |
化學蛋白質體學是一門新興的研究領域,主要目的在於鑑定與驗證活性小分子之標的蛋白質,利用化合物與蛋白質間特異性的相互作用力成為一個強而有力的搜尋技術。本篇論文目的為在人類急性骨髓性白血病(AML)細胞株MV4-11中找到sunitinib之標的蛋白質。先前文獻中顯示出小分子藥物sunitinib有抑制AML細胞株中FLT3磷酸化的作用,進而有效地抑制掉MV4-11的增殖組成。利用化學蛋白質體學的策略,sunitinib合成的生物素標記藥物固定於鏈黴卵白素粒子上形成親和性管柱,建立活性為基礎的探針於MV4-11細胞株中抓取特定蛋白質。實驗設計中導入negative控制組,其結構與biotinylated sunitinib相似但不具有藥理活性,做為背景值來減少非特異性蛋白質之鍵結。經過親和性層析適當的洗滌與洗脫過程,被洗脫之蛋白質以一維膠體電泳(SDS-PAGE)蛋白質和液相層析串聯式質譜儀(LC-MS/MS)進行分析和蛋白質資料庫比對鑑定出候選標的蛋白質。MV4-11細胞裂解液中有248個標的蛋白質被鑑定到;他們大部分可能為sunitinib之新奇作用物和標的蛋白質。藉由GeneGo路徑分析,在muscle contraction relaxin和development VEGF signaling via VEGFR2 generic cascades這兩項路徑中,RAP-1A和MAP2K1被認為是藥物目標物而且他們與血管的生成有關係,造成影響MV4-11細胞增殖與存活的原因。
Chemical proteomics is an emerging field that the goal is to identify and validate target proteins for bioactive small molecules. It utilizes the specific interaction of compound and the target proteins to be a powerful screening technology. The main purpose of this study is to identify sunitinib target proteins in acute myeloid leukemia (MV4-11) cell lines. It has been reported that sunitinib inhibits phosphorylation of FLT3 in AML cell lines, which potently inhibits constitutive proliferation of MV4-11. Using strategy of chemical proteomics, the synthesized biotin-tagged sunitinib was immobilized on streptavidin beads to make affinity column as an activity-based probe that captures the target proteins in MV4-11 cells. A negative control, which structure was similar to biotinylated sunitinib with no pharmacological activity, was designed as a background for eliminating the impact resulting from non-specific binding proteins. After appropriate wash and elution steps in the affinity chromatography, the eluted proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and liquid chromatography tandem mass spectrometry (LC-MS/MS). Two hundred forty eight sunitinib-binding proteins were identified in MV4-11 cell lysate. They are potential interactors and targets of sunitinib. These sunitinib-binding proteins were further analyzed by GeneGo and found they were associated with muscle contraction relaxing and development VEGF signaling via VEGFR2 cascades signaling pathways. Among them, RAP-1A and MAP2K1 are considered drug targets and related to angiogenesis. They are also contributed to the effects on MV4-11 cell proliferation and survival.
(1) Goodman, V. L.; Rock, E. P.; Dagher, R.; Ramchandani, R. P.; Abraham, S.; Gobburu, J. V.; Booth, B. P.; Verbois, S. L.; Morse, D. E.; Liang, C. Y.; Chidambaram, N.; Jiang, J. X.; Tang, S.; Mahjoob, K.; Justice, R.; Pazdur, R. Clinical cancer research : an official journal of the American Association for Cancer Research 2007, 13, 1367.
(2) Rock, E. P.; Goodman, V.; Jiang, J. X.; Mahjoob, K.; Verbois, S. L.; Morse, D.; Dagher, R.; Justice, R.; Pazdur, R. The oncologist 2007, 12, 107.
(3) Chow, L. Q.; Eckhardt, S. G. Journal of clinical oncology : official journal of the American Society of Clinical Oncology 2007, 25, 884.
(4) Izzedine, H.; Buhaescu, I.; Rixe, O.; Deray, G. Cancer Chemother Pharmacol 2007, 60, 357.
(5) Shukla, S.; Robey, R. W.; Bates, S. E.; Ambudkar, S. V. Drug metabolism and disposition: the biological fate of chemicals 2009, 37, 359.
(6) Dasanu, C. A.; Alexandrescu, D. T.; Dutcher, J. Southern medical journal 2007, 100, 328.
(7) Motzer, R. J.; Hutson, T. E.; Tomczak, P.; Michaelson, M. D.; Bukowski, R. M.; Rixe, O.; Oudard, S.; Negrier, S.; Szczylik, C.; Kim, S. T.; Chen, I.; Bycott, P. W.; Baum, C. M.; Figlin, R. A. The New England journal of medicine 2007, 356, 115.
(8) Jemal, A.; Thomas, A.; Murray, T.; Thun, M. CA: a cancer journal for clinicians 2002, 52, 23.
(9) Le Beau, M. M.; Albain, K. S.; Larson, R. A.; Vardiman, J. W.; Davis, E. M.; Blough, R. R.; Golomb, H. M.; Rowley, J. D. Journal of clinical oncology : official journal of the American Society of Clinical Oncology 1986, 4, 325.
(10) Austin, H.; Delzell, E.; Cole, P. American journal of epidemiology 1988, 127, 419.
(11) Thirman, M. J.; Gill, H. J.; Burnett, R. C.; Mbangkollo, D.; McCabe, N. R.; Kobayashi, H.; Ziemin-van der Poel, S.; Kaneko, Y.; Morgan, R.; Sandberg, A. A.; et al. The New England journal of medicine 1993, 329, 909.
(12) Bizzozero, O. J., Jr.; Johnson, K. G.; Ciocco, A. The New England journal of medicine 1966, 274, 1095.
(13) Sanz, G. F.; Sanz, M. A.; Vallespi, T.; Canizo, M. C.; Torrabadella, M.; Garcia, S.; Irriguible, D.; San Miguel, J. F. Blood 1989, 74, 395.
(14) Horwitz, M. Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K 1997, 11, 1347.
(15) Rohatiner, A. Z.; Gregory, W. M.; Bassan, R.; Barnett, M. J.; Waxman, J.; Richards, M. A.; Ganesan, T. S.; Tucker, J.; Malik, S. T.; Biruls, R.; et al. Journal of clinical oncology : official journal of the American Society of Clinical Oncology 1988, 6, 218.
(16) Preisler, H. D.; Raza, A.; Early, A.; Kirshner, J.; Brecher, M.; Freeman, A.; Rustum, Y.; Azarnia, N.; Priore, R.; Sandberg, A.; et al. Journal of clinical oncology : official journal of the American Society of Clinical Oncology 1987, 5, 722.
(17) Cassileth, P. A.; Begg, C. B.; Silber, R.; Spiers, A.; Burkart, P. T.; Scharfman, W.; Knospe, W. H.; Bennett, J. M.; Mazza, J. J.; Oken, M. M.; et al. Cancer treatment reports 1987, 71, 137.
(18) Zittoun, R.; Jehn, U.; Fiere, D.; Haanen, C.; Lowenberg, B.; Willemze, R.; Abels, J.; Bury, J.; Peetermans, M.; Hayat, M.; et al. Blood 1989, 73, 896.
(19) Small, D.; Levenstein, M.; Kim, E.; Carow, C.; Amin, S.; Rockwell, P.; Witte, L.; Burrow, C.; Ratajczak, M. Z.; Gewirtz, A. M.; et al. Proceedings of the National Academy of Sciences of the United States of America 1994, 91, 459.
(20) Yamamoto, Y.; Kiyoi, H.; Nakano, Y.; Suzuki, R.; Kodera, Y.; Miyawaki, S.; Asou, N.; Kuriyama, K.; Yagasaki, F.; Shimazaki, C.; Akiyama, H.; Saito, K.; Nishimura, M.; Motoji, T.; Shinagawa, K.; Takeshita, A.; Saito, H.; Ueda, R.; Ohno, R.; Naoe, T. Blood 2001, 97, 2434.
(21) Lander, E. S.; Linton, L. M.; Birren, B.; Nusbaum, C.; Zody, M. C.; Baldwin, J.; Devon, K.; Dewar, K.; Doyle, M.; FitzHugh, W.; Funke, R.; Gage, D.; Harris, K.; Heaford, A.; Howland, J.; Kann, L.; Lehoczky, J.; LeVine, R.; McEwan, P.; McKernan, K.; Meldrim, J.; Mesirov, J. P.; Miranda, C.; Morris, W.; Naylor, J.; Raymond, C.; Rosetti, M.; Santos, R.; Sheridan, A.; Sougnez, C.; Stange-Thomann, N.; Stojanovic, N.; Subramanian, A.; Wyman, D.; Rogers, J.; Sulston, J.; Ainscough, R.; Beck, S.; Bentley, D.; Burton, J.; Clee, C.; Carter, N.; Coulson, A.; Deadman, R.; Deloukas, P.; Dunham, A.; Dunham, I.; Durbin, R.; French, L.; Grafham, D.; Gregory, S.; Hubbard, T.; Humphray, S.; Hunt, A.; Jones, M.; Lloyd, C.; McMurray, A.; Matthews, L.; Mercer, S.; Milne, S.; Mullikin, J. C.; Mungall, A.; Plumb, R.; Ross, M.; Shownkeen, R.; Sims, S.; Waterston, R. H.; Wilson, R. K.; Hillier, L. W.; McPherson, J. D.; Marra, M. A.; Mardis, E. R.; Fulton, L. A.; Chinwalla, A. T.; Pepin, K. H.; Gish, W. R.; Chissoe, S. L.; Wendl, M. C.; Delehaunty, K. D.; Miner, T. L.; Delehaunty, A.; Kramer, J. B.; Cook, L. L.; Fulton, R. S.; Johnson, D. L.; Minx, P. J.; Clifton, S. W.; Hawkins, T.; Branscomb, E.; Predki, P.; Richardson, P.; Wenning, S.; Slezak, T.; Doggett, N.; Cheng, J. F.; Olsen, A.; Lucas, S.; Elkin, C.; Uberbacher, E.; Frazier, M. Nature 2001, 409, 860.
(22) Reiss, T. Trends in biotechnology 2001, 19, 496.
(23) Venter, J. C.; Adams, M. D.; Myers, E. W.; Li, P. W.; Mural, R. J.; Sutton, G. G.; Smith, H. O.; Yandell, M.; Evans, C. A.; Holt, R. A.; Gocayne, J. D.; Amanatides, P.; Ballew, R. M.; Huson, D. H.; Wortman, J. R.; Zhang, Q.; Kodira, C. D.; Zheng, X. H.; Chen, L.; Skupski, M.; Subramanian, G.; Thomas, P. D.; Zhang, J.; Gabor Miklos, G. L.; Nelson, C.; Broder, S.; Clark, A. G.; Nadeau, J.; McKusick, V. A.; Zinder, N.; Levine, A. J.; Roberts, R. J.; Simon, M.; Slayman, C.; Hunkapiller, M.; Bolanos, R.; Delcher, A.; Dew, I.; Fasulo, D.; Flanigan, M.; Florea, L.; Halpern, A.; Hannenhalli, S.; Kravitz, S.; Levy, S.; Mobarry, C.; Reinert, K.; Remington, K.; Abu-Threideh, J.; Beasley, E.; Biddick, K.; Bonazzi, V.; Brandon, R.; Cargill, M.; Chandramouliswaran, I.; Charlab, R.; Chaturvedi, K.; Deng, Z.; Di Francesco, V.; Dunn, P.; Eilbeck, K.; Evangelista, C.; Gabrielian, A. E.; Gan, W.; Ge, W.; Gong, F.; Gu, Z.; Guan, P.; Heiman, T. J.; Higgins, M. E.; Ji, R. R.; Ke, Z.; Ketchum, K. A.; Lai, Z.; Lei, Y.; Li, Z.; Li, J.; Liang, Y.; Lin, X.; Lu, F.; Merkulov, G. V.; Milshina, N.; Moore, H. M.; Naik, A. K.; Narayan, V. A.; Neelam, B.; Nusskern, D.; Rusch, D. B.; Salzberg, S.; Shao, W.; Shue, B.; Sun, J.; Wang, Z.; Wang, A.; Wang, X.; Wang, J.; Wei, M.; Wides, R.; Xiao, C.; Yan, C. Science 2001, 291, 1304.
(24) Wasinger, V. C.; Cordwell, S. J.; Cerpa-Poljak, A.; Yan, J. X.; Gooley, A. A.; Wilkins, M. R.; Duncan, M. W.; Harris, R.; Williams, K. L.; Humphery-Smith, I. Electrophoresis 1995, 16, 1090.
(25) Blackstock, W. P.; Weir, M. P. Trends in biotechnology 1999, 17, 121.
(26) Wheeler, C. H.; Berry, S. L.; Wilkins, M. R.; Corbett, J. M.; Ou, K.; Gooley, A. A.; Humphery-Smith, I.; Williams, K. L.; Dunn, M. J. Electrophoresis 1996, 17, 580.
(27) Washburn, M. P.; Wolters, D.; Yates, J. R., 3rd Nature biotechnology 2001, 19, 242.
(28) Wolters, D. A.; Washburn, M. P.; Yates, J. R., 3rd Analytical chemistry 2001, 73, 5683.
(29) Swanson, S. K.; Washburn, M. P. Drug discovery today 2005, 10, 719.
(30) Persidis, A. Nature biotechnology 1998, 16, 1082.
(31) Han, S. Y.; Hwan Kim, S. Archiv der Pharmazie 2007, 340, 169.
(32) Duvel, J.; Bertinetti, D.; Moller, S.; Schwede, F.; Morr, M.; Wissing, J.; Radamm, L.; Zimmermann, B.; Genieser, H. G.; Jansch, L.; Herberg, F. W.; Haussler, S. Journal of microbiological methods 2012, 88, 229.
(33) Liu, Y.; Patricelli, M. P.; Cravatt, B. F. Proceedings of the National Academy of Sciences of the United States of America 1999, 96, 14694.
(34) Deu, E.; Yang, Z.; Wang, F.; Klemba, M.; Bogyo, M. PloS one 2010, 5, e11985.
(35) Adam, G. C.; Sorensen, E. J.; Cravatt, B. F. Molecular & cellular proteomics : MCP 2002, 1, 828.
(36) Verhelst, S. H.; Bogyo, M. BioTechniques 2005, 38, 175.
(37) Tamura, T.; Terada, T.; Tanaka, A. Bioconjugate chemistry 2003, 14, 1222.
(38) Shiyama, T.; Furuya, M.; Yamazaki, A.; Terada, T.; Tanaka, A. Bioorganic & medicinal chemistry 2004, 12, 2831.
(39) Sakamoto, S.; Hatakeyama, M.; Ito, T.; Handa, H. Bioorganic & medicinal chemistry 2012, 20, 1990.
(40) Kidd, D.; Liu, Y.; Cravatt, B. F. Biochemistry 2001, 40, 4005.
(41) Patricelli, M. P.; Giang, D. K.; Stamp, L. M.; Burbaum, J. J. Proteomics 2001, 1, 1067.
(42) Greenbaum, D.; Medzihradszky, K. F.; Burlingame, A.; Bogyo, M. Chemistry & biology 2000, 7, 569.
(43) Kato, D.; Boatright, K. M.; Berger, A. B.; Nazif, T.; Blum, G.; Ryan, C.; Chehade, K. A.; Salvesen, G. S.; Bogyo, M. Nature chemical biology 2005, 1, 33.
(44) Li, Y. M.; Xu, M.; Lai, M. T.; Huang, Q.; Castro, J. L.; DiMuzio-Mower, J.; Harrison, T.; Lellis, C.; Nadin, A.; Neduvelil, J. G.; Register, R. B.; Sardana, M. K.; Shearman, M. S.; Smith, A. L.; Shi, X. P.; Yin, K. C.; Shafer, J. A.; Gardell, S. J. Nature 2000, 405, 689.
(45) Chan, E. W.; Chattopadhaya, S.; Panicker, R. C.; Huang, X.; Yao, S. Q. Journal of the American Chemical Society 2004, 126, 14435.
(46) Saghatelian, A.; Jessani, N.; Joseph, A.; Humphrey, M.; Cravatt, B. F. Proceedings of the National Academy of Sciences of the United States of America 2004, 101, 10000.
(47) Sieber, S. A.; Niessen, S.; Hoover, H. S.; Cravatt, B. F. Nature chemical biology 2006, 2, 274.
(48) Hekmat, O.; Kim, Y. W.; Williams, S. J.; He, S.; Withers, S. G. The Journal of biological chemistry 2005, 280, 35126.
(49) Salisbury, C. M.; Cravatt, B. F. Proceedings of the National Academy of Sciences of the United States of America 2007, 104, 1171.
(50) Adam, G. C.; Sorensen, E. J.; Cravatt, B. F. Nature biotechnology 2002, 20, 805.
(51) Barglow, K. T.; Cravatt, B. F. Chemistry & biology 2004, 11, 1523.
(52) Lee, W. C.; Lee, K. H. Analytical biochemistry 2004, 324, 1.
(53) Sleno, L.; Emili, A. Current opinion in chemical biology 2008, 12, 46.
(54) Jonker, N.; Kool, J.; Irth, H.; Niessen, W. M. Analytical and bioanalytical chemistry 2011, 399, 2669.
(55) Harding, M. W.; Galat, A.; Uehling, D. E.; Schreiber, S. L. Nature 1989, 341, 758.
(56) Taunton, J.; Hassig, C. A.; Schreiber, S. L. Science 1996, 272, 408.
(57) Fernbach, N. V.; Planyavsky, M.; Muller, A.; Breitwieser, F. P.; Colinge, J.; Rix, U.; Bennett, K. L. Journal of proteome research 2009, 8, 4753.
(58) Frick, W.; Bauer-Schafer, A.; Bauer, J.; Girbig, F.; Corsiero, D.; Heuer, H.; Kramer, W. Bioorganic & medicinal chemistry 2003, 11, 1639.
(59) Lee, J. S.; Han, S. Y.; Kim, M. S.; Yu, C. M.; Kim, M. H.; Kim, S. H.; Min, Y. K.; Kim, B. T. Bioorganic & medicinal chemistry letters 2006, 16, 4733.
(60) Raymond, S.; Weintraub, L. Science 1959, 130, 711.
(61) Fenn, J. B.; Mann, M.; Meng, C. K.; Wong, S. F.; Whitehouse, C. M. Science 1989, 246, 64.
(62) Emmett, M. R.; White, F. M.; Hendrickson, C. L.; Shi, S. D.; Marshall, A. G. Journal of the American Society for Mass Spectrometry 1998, 9, 333.
(63) Yi, E. C.; Lee, H.; Aebersold, R.; Goodlett, D. R. Rapid communications in mass spectrometry : RCM 2003, 17, 2093.
(64) Yates, J. R., 3rd; Eng, J. K.; McCormack, A. L.; Schieltz, D. Analytical chemistry 1995, 67, 1426.
(65) Mortz, E.; O'Connor, P. B.; Roepstorff, P.; Kelleher, N. L.; Wood, T. D.; McLafferty, F. W.; Mann, M. Proceedings of the National Academy of Sciences of the United States of America 1996, 93, 8264.
(66) Kancha, R. K.; Grundler, R.; Peschel, C.; Duyster, J. Experimental hematology 2007, 35, 1522.
(67) Fenton, M. S.; Marion, K. M.; Salem, A. K.; Hogen, R.; Naeim, F.; Hershman, J. M. Thyroid : official journal of the American Thyroid Association 2010, 20, 965.
(68) Padhye, S. S.; Guin, S.; Yao, H. P.; Zhou, Y. Q.; Zhang, R.; Wang, M. H. Molecular pharmaceutics 2011, 8, 2310.