Title

Pt(1R, 2R-dach)22+與DNA結合之序列偏好

Translated Titles

Sequence preferences of Pt(1R, 2R-dach)22+

Authors

游岳寧

Key Words

DNA結合 ; Pt(1R, 2R-dach)22+ ; 順鉑 ; DNA binding ; Pt(1R, 2R-dach)22+ ; cisplatin

PublicationName

臺灣師範大學化學系學位論文

Volume or Term/Year and Month of Publication

2010年

Academic Degree Category

碩士

Advisor

黃文彰

Content Language

繁體中文

Chinese Abstract

本研究係針對 cis-Pt(NH3)2Cl2 (cisplatin) 的衍生物, Pt(1R, 2R-dach)22+ (dach:diaminocyclohexane) ,探討其與cisplatin 反應途徑之異同,與反應條件及序列之偏好。 經由與cisplatin相同條件實驗結果,顯示 Pt(1R, 2R-dach)22+ 不會如 cisplatin 之反應途徑,與 DNA 產生 cis-GG 的交聯鍵結反應。 根據 UV 及 CD 升溫圖譜研究顯示 Pt(1R, 2R-dach)22+ 可與 AT-rich 之 DNA 序列產生溝槽結合反應。反應條件溫度需 40℃ , Pt(1R, 2R-dach)22+:DNA則需在 30:1 以上,為本實驗可產生反應之必要條件。 由本實驗 10 條 DNA 序列之 UV 及 CD 升溫圖譜分析,Pt(1R, 2R-dach)22+ 對含單結合位置的 DNA 序列,其結合強度皆不強,而對含雙結合位置的 DNA 序列則都近乎完全反應,推測d(CXXCGXXG)2 及d(CXXCCGGXXG)2 (X=A或T) 為 Pt(1R, 2R-dach)22+ 偏好之 DNA 序列特徵,為至少 2 組連續 A 或 T 的鹼基對,此協同性結合(cooperative binding) 為正協同作用 (positive cooperativity)。

English Abstract

This investigation was carried out via a cis-Pt(NH3)2Cl2 (cisplatin) derivative, Pt(1R, 2R-dach)22+ (dach:diaminocyclohexane), and focused on the different reaction pathways, reaction conditions and sequence preferences between cisplatin and Pt(1R, 2R-dach)22+. Under the same experiment conditions as cisplatin, the results suggested Pt(1R, 2R-dach)22+ do not follow the same mechanism as cisplatin, in forming cis-GG crosslink with DNA. Based on variable temperature UV and CD denaturation analysis, Pt(1R, 2R-dach)22+ can react with AT-rich DNA sequences through groove binding. Based on experiments carried out under various conditions, suggested that reaction temperature about 40℃ and Pt(1R, 2R-dach)22+:DNA over 30:1 were required to promote the reaction. Refer to UV and CD denaturation experiment of ten DNA samples of variable sequences in this work, Pt(1R, 2R-dach)22+ has only weak binding affinity to single binding site sequences, whereas double binding site sequences react almost completely with Pt(1R, 2R-dach)22+. The variable sequence studies suggested that the consensus sequences of Pt(1R, 2R-dach)22+-DNA sites are d(CXXCGXXG)2 and d(CXXCCGGXXG)2 (X=A or T) with at least 2 base pairs of A or T.

Topic Category 基礎與應用科學 > 化學
理學院 > 化學系
Reference
  1. Takahara, P. M.; Frederick, C. A.; Lippard, S. J. J. Am. chem. Soc. 1996, 118, 12309-12321.
    連結:
  2. Jamieson, E. R.; Lippard, S. J. Chem. Rev. 1999, 99, 2467-2498.
    連結:
  3. Bernges, F.; Holler E. Nucleic Acids Res. 1991, Vol. 19, No. 7, 1483-1489.
    連結:
  4. Drew, H. D. K. J. Chem. Soc. 1932, 2328-2331.
    連結:
  5. Wheat, N. J.; Collins, J. G. J. Inorg. Biochem. 2000, 78, 313-320.
    連結:
  6. Nelson, D. L.; Cox, M. M. (2004) Lehninger: Principles of Biochemistry (Fourth Ed.) W. H. Freeman and Company, New York, chapter 8.
    連結:
  7. Fasman, G. D. (1975) CRC Handbook of Biochemistry and Molecular Biology (Third Ed.), chapter I, P.589.
    連結:
  8. Mergny, J.-L.; Lacroix, L. Oligonucleotides 2003, 13, 515-537.
    連結:
  9. 蔡惠如,國立台灣師範大學化學研究所(民95)
  10. Jung, Y.; Lippard, S. J. Chem. Rev. 2007, Vol. 107, No. 5, 1387-1407.
  11. Wong, E.; Giandomenico, C. M. Chem. Rev. 1999, 99, 2451-2466.
  12. Yang, D.; Wang, A. H.-J. Prog. Biophys. molec. Biol. 1996, Vol. 66, No. 1, 81-111.
  13. Pendyala, L.; Kidani, Y.; Perez, R.; Wilkes, J.; Bemacki, R. J.; Creaven, P. J. Cancer Letters 1995, 97, 117-184.
  14. Franklin, C. A.; Fry, J. V.; Collins, J. G. J. Inorg. Biochem. 1996, 35, 7541-7545.
  15. Fede, A.; Labhardt, A.; Bannwarth, W.; Leupin, W. Biochem. 1991, 30, 11377-11388.
  16. Watts, C. R.; Kerwin, S. M.; Kenyon, G. L.; Kuntz, I. D.; Kallick, D. A. J. Am. Chem. Soc. 1995, 117, 9941-9950.
  17. Baleja, J. D.; Pon, R. T.; Sykes, B. D. Biochemistry 1990, 29, 4828-4839.
  18. Eyring, H.; Liu, H.-C.; Caldwell, D. Chem. Rev. 1968, 68, 525-540.
  19. Shi, S.; Yan, L.; Yang, Y.; Fisher-Shaulsky, J.; Thacher, T. J. Comput.Chem. 2003, Vol. 24, No. 9, 1059-1076.
  20. Poklar, N.; Pilch, D. S.; Lippard, S. J.; Redding, E. A.; Dunham, S. U.; Breslauer, K. J. Proc. Natl. Acad. Sci. U. S. A. 1996, 93, 7606-7611.
  21. Nina, B.; Koji, N,; Robert, W. W. (2000) Circular Dichroism Principles and Applications (Second Ed.), P.707.