DNA作为遗传信息的载体，了解在细胞核内的组装和结构具有非常重要的意义。目前，使用能够与DNA结合的细胞膜渗透性良好的有机荧光分子作为DNA标记探针是实现这一目标的主要手段。自Barton实验室发现阳离子Ru（Ⅱ）络合物[Ru（bpy）2（dppz）]2+（bpy=2，2'-联吡啶，dppz=多吡啶并吩嗪）能够作为DNA分子"光开关"以来，d6八面体多吡啶金属络合物与DNA的结合特性及相关研究，尤其是对这类金属络合物作为高灵敏度和结构特异性DNA探针的研究，便吸引了人们极大的关注。由于这类物质较低的膜渗透性，先前多数研究都只是局限于细胞外，活细胞内DNA直接显像成功的实例较为少见。本课题组发现五氯酚（PCP）和另外两类结构不相关的生化试剂能促进Ru（Ⅱ）多吡啶络合物的细胞特别是细胞核的摄取，[Ru（bpy）2（dppz）]2+/PCP之间的协同核摄取机理可能是形成了较稳定的亲脂性离子对复合物。Ru（Ⅱ）络合物的两种对映异构体在活细胞内与DNA结合后表现出了明显的手性选择性。这是我们首次发现通过以形成离子对复合物的方式将DNA分子"光开关"Ru（Ⅱ）络合物转入活细胞核内并维持其"光开关"效应，可为研究将其他潜在的具有生物医疗效应的细胞膜不通透金属络合物转入细胞内提供一种全新的方法。
Since the discovery of DNA as the genetic material carrier, the work towards the elucidation of DNA structure within the cell nucleus has become of great importance. Fluorescent microscopy using luminescent and cell membrane permeable organic DNA-binding molecule as probes is a well-established technique towards achieving this goal. Barton's group discovered that the cationic ruthenium complex[Ru(bpy)2(dppz)]2+ (bpy=2,2'-bipyridine, dppz=dipyrido[3,2-a:2',3'-c] phenazine) functions as a molecular ‘light switch’ for DNA. Since then, there has been great attention drawn to the DNA binding properties of polypyridyl complexes of d6 octahedral metal ions, specifically towards the development of highly sensitive and structure-specific DNA probes.Until recently the research has been largely focused on the development of in vitro probes. However, few studies involving direct imaging of DNA in live cells with such systems have had very limited success, with poor membrane permeability still being ascribed as the major limiting factor. We found that not only the cellular, but more interestingly and importantly, the nuclear uptake of[Ru(bpy)2(dppz)]2+ is remarkably enhanced by pentachlorophenol and two other structurally unrelated biochemical agents. Furthermore, enantioselective imaging of live-cell nuclear DNA is observed between the two chiral forms of Ru(Ⅱ) complexes. The underlying molecular mechanism is found to be the formation of novel lipophilic and relatively stable ion-pair complexes. This represents the first report for an unprecedented new method for delivering the DNA ‘light-switching’ Ru(Ⅱ) complexes into the nucleus of living cells via ion-pairing, which could serve as a promising general live-cell delivering method for other potentially bio-medically but cell-impermeable metal complexes.
Contents
1 Introduction
2 Predominant interactions between metal chelates and DNA
2.1 Irreversible binding with DNA
2.2 Reversible binding with DNA
3 Binding with intracellular DNA
3.1 Cellular uptake mechanism
3.2 Potential luminescent probe for intracellular DNA
3.3 Hydrophobicity and cellular uptake regulation
3.4 Appending targeting group for cellular uptake promotion
3.5 Active transport of chelates
3.6 Enhanced cellular uptake of “DNA light switch” Ru(Ⅱ) complex via forming lipophilic ion-pairing complexes
4 Methods for assessing cellular uptake
4.1 Transmission electron microscope
4.2 Raman spectra
4.3 Molecular labeling methods with fluorophore
4.4 Methods for quantitative uptake measurements
5 Cytotoxicity
6 The medical value for Ru(Ⅱ) polypyridyl complexes
[1] Watson J D, Crick F H C. Nature, 1953, 171(4356):737.
[2] Saegner W. Principles of Nucleic Acid Structure. Springer-Verlag NY:Springer, 1984.
[3] Neidle S. Nucleic Acid Structure and Recognition.Oxford UK:Oxford University Press, 2002.
[4] Kumar C V, Barton J K, Turro N J. J. Am. Chem. Soc., 1985, 107(19):5518.
[5] Zeglis B M, Pierre V C, Barton J K. Chem. Commun., 2007, (44):4565.
[6] Zhang C X, Lippard S J. Curr. Opin. Chem. Biol., 2003, 7(4):481.
[7] Hartinger C G, Dyson P J. Chem. Soc. Rev., 2009, 38(2):391.
[8] Ott I, Gust R. Arch. Pharm., 2007, 340(3):117.
[9] Bruijnincx P C A, Sadler P J. Curr. Opin. Chem. Biol., 2008, 12(2):197.
[10] Jakupec M A, Galanski M, Arion V B, Hartinger C G, Keppler B K. Dalton Trans., 2008, (2):183.
[11] Dyson P J, Sava G. Dalton Trans., 2006, (16):1929.
[12] Allardyce C S, Dyson P J. Platin. Met. Rev., 2001, 45(2):62.
[13] Depenbrock H, Schmelcher S, Peter R, Keppler B K, Weirich G, Block T, Rastetter J, Hanauske A R. Eur. J. Cancer., 1997, 33(14):2404.
[14] Bergamo A, Gava B, Alessio E, Mestroni G, Serli B, Cocchietto M,Zorzet S, Sava G. Int. J. Oncol., 2002, 21(6):1331.
[15] Alessio E, Mestroni G, Bergamo A, Sava G. Curr. Top. Med. Chem., 2004, 4(15):1525.
[16] Hartinger C G, Zorbas-Seifried S,Jakupec M A,Kynast B, Zorbas H, Keppler B K. J. Inorg. Biochem., 2006, 100(5/6):891.
[17] Cohen G L, Bauer W R, Barton J K, Lippard S J. Science, 1979, 203(4384):1014.
[18] Mei H Y, Barton J K. Proc. Natl. Acad. Sci. U.S.A., 1988, 85(5):1339.
[19] Brabec V. Prog. Nucleic. Acid. Re., 2002, 71:1.
[20] Kastan M B, Bartek J. Nature, 2004, 432(7015):316.
[21] Gelasco A, Lippard S J. Biochemistry, 1998, 37(26):9230.
[22] Takahara P M, Rosenzweig A C, Frederick C A, Lippard S J. Nature, 1995, 377(6550):649.
[23] Clarke M J, Zhu F C, Frasca D R. Chem. Rev., 1999, 99(9):2511.
[24] Metcalfe C, Thomas J A. Chem. Soc. Rev., 2003, 32(4):215.
[25] Shi Y, Guo C L, Sun Y J, Liu Z L, Xu F G, Zhang Y, Wen Z W, Li Z A. Biomacromolecules, 2011, 12(3):797.
[26] Erkkila K E, Odom D T, Barton J K. Chem. Rev., 1999, 99(9):2777.
[27] Barton J K, Danishefsky A T, Goldberg J M. J. Am. Chem. Soc., 1984, 106(7):2172.
[28] Barton J K, Goldberg J M, Kumar C V, Turro N J. J. Am. Chem. Soc., 1986, 108(8):2081.
[29] Barton J K, Raphael A L. Proc. Natl. Acad. Sci. U.S.A., 1985, 82(19):6460.
[30] Hudson B P, Barton J K. J. Am. Chem. Soc., 1998, 120(28):6877.
[31] Franklin S J, Barton J K. Biochemistry, 1998, 37(46):16093.
[32] Kielkopf C L, Erkkila K E, Hudson B P, Barton J K, Rees D C. Nat. Struct. Biol., 2000, 7(2):117.
[33] Friedman A E, Chambron J C, Sauvage J P, Turro N J, Barton J K. J. Am. Chem. Soc., 1990, 112(12):4960.
[34] Hartshorn R M, Barton J K. J. Am. Chem. Soc., 1992, 114(15):5919.
[35] Amouyal E, Homsi A, Chambro J C. Sauvage J P. Dalton Trans., 1990, (6):1841.
[36] Turro C, Bossmann S H, Jenkins Y, Barton J K, Turro N J. J. Am. Chem. Soc., 1995, 117(35):9026.
[37] Satyanarayana S, Dabrowiak J C, Chaires J B. Biochemistry, 1993, 32(10):2573.
[38] Coggan D Z M, Haworth I S, Bates P J, Robinson A, Rodger A. Inorg. Chem., 1999, 38(20):4486.
[39] Foley F M, Keene F R, Collins J G. Dalton Trans., 2001, (20):2968.
[40] Pascu G I, Hotze A C, Sanchez-Cano C, Kariuki B M, Hannon M J. Angew. Chemie., 2007, 46(23):4374.
[41] Glasson C R K, Meehan G V, Clegg J K, Lindoy L F, Smith J A, Keene F R, Motti C. Chem. Eur. J., 2008, 14(34):10535.
[42] Gonzalez V, Wilson T, Kurihara I, Imai A, Thomas J A, Otsuki J. Chem. Commun., 2008, (16):1868.
[43] Lutterman D A, Chouai A, Liu Y, Sun Y, Stewart C D, Dunbar K R, Turro C. J. Am. Chem. Soc., 2008, 130(4):1163.
[44] Ghosh A, Das P, Gill M R, Kar P,Walker M G, Thomas J A, Das A. Chem. Eur. J., 2011, 17(7):2089.
[45] Hiort C, Lincoln P, Nordén B. J. Am. Chem. Soc., 1993, 115(9):3448.
[46] Yun B H, Kim J O, Lee B W, Lincoln P, Norden B, Kim J M, Kim S K. J. Phys. Chem. B, 2003, 107(36):9858.
[47] Biver T, Cavazza C, Secco F, Venturini M. J. Inorg. Biochem., 2007, 101(3):461.
[48] McKinley A W, Lincoln P, Tuite E M. Coordi. Chem. Rev., 2011, 255(21/22):2676.
[49] Keene F R, Smith J A, Collins J G. Coordi. Chem. Rev.,2009, 253(15/16):2021.
[50] Gill M R, Thomas J A. Chem. Soc. Rev., 2012, 41(8):3179.
[51] Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P. Molecular Biology of the Cell, 5th ed.NY:Garland Science, 2007.
[52] Barton J K, Basile L A, Danishefsky A, Alexandrescu A. Proc. Natl. Acad. Sci. U.S.A., 1984, 81(7):1961.
[53] Jackson B A, Barton J K. Biochemistry, 2000, 39(20):6176.
[54] Junicke H, Hart J R, Kisko J, Glebov O, Kirsch I R, Barton J K. Proc. Natl. Acad. Sci. U.S.A., 2003, 100(7):3737.
[55] Ruba E, Hart J R, Barton J K. Inorg. Chem., 2004, 43(15):4570.
[56] Lim M H, Song H, Olmon E D, Dervan E E, Barton J K. Inorg. Chem., 2009, 48(12):5392.
[57] Pierre V C, Kaiser J T, Barton J K. Proc. Natl. Acad. Sci. U.S.A., 2007, 104(2):429.
[58] Zeglis B M, Pierre V C, Kaiser J T, Barton J K. Biochemistry., 2009, 48(20):4247.
[59] Patterson B T, Collins J G, Foley F M, Keene F R. Dalton Trans., 2002, (23):4343.
[60] Smith J A, Collins J G, Patterson B T, Keene F R. Dalton Trans., 2004, (9):1277.
[61] Smith J A, Keene F R. Chem. Commun., 2006, (24):2583.
[62] Jimenez-Hernandez M E, Orellana G, Montero F, Portoles M T. Photochem. Photobiol., 2000, 72(1):28.
[63] New E J, Parker D, Smith D G, Walton J W. Curr. Opin. Chem. Biol., 2010, 14(2):238.
[64] Fernandez-Moreira V, Thorp-Greenwood F L, Coogan M P. Chem. Commun., 2010, 46(2):186.
[65] Zhao Q, Huang C H, Li F Y. Chem. Soc. Rev., 2011, 40(5):2508.
[66] Onfelt B, Gostring L, Lincoln P, Norden B, Onfelt A. Mutagenesis, 2002, 17(4):317.
[67] Puckett C A, Barton J K. J. Am. Chem. Soc., 2007, 129(1):46.
[68] Puckett C A, Barton J K. Biochemistry, 2008, 47(45):11711.
[69] Rajendiran V, Palaniandavar M, Periasamy V S, Akbarsha M A. J. Inorg. Biochem., 2010, 104(2):217.
[70] Amoroso A J, Coogan M P, Dunne J E, Fernandez-Moreira V, Hess J B, Hayes, A J, Lloyd D, Millet C, Pope S J A, Williams C. Chem. Commun., 2007, (29):3066.
[71] Lo K K W, Lee T K M, Lau J S Y, Poon W L, Cheng S H. Inorg. Chem., 2008, 47(1):200.
[72] Neugebauer U, Pellegrin Y, Devocelle M, Forster R J, Signac W, Morand N, Keyes T E. Chem. Commun., 2008, (42):5307.
[73] Brunner J, Barton J K. Biochemistry, 2006, 45(40):12295.
[74] Puckett C A, Barton J K. J. Am. Chem. Soc., 2009, 131(25):8738.
[75] Svensson F R, Matson M, Li M, Lincoln P. Biophys. Chem., 2010, 149(3):102.
[76] Matson M, Svensson F R, Norden B, Lincoln P. J. Phys. Chem. B, 2011, 115(7):1706.
[77] Svensson F R, Abrahamsson M, Stromberg N, Ewing A G, Lincoln P. J. Phys. Chem. Lett., 2011, 2(5):397.
[78] Puckett C A, Barton J K. Bioorg. Med. Chem., 2010, 18(10):3564.
[79] Gottschaldt M, Schubert U S, Rau S, Yano S, Vos J G, Kroll T, Clement J, Hilger I. ChemBioChem, 2010, 11(5):649.
[80] Gill M R, Garcia-Lara J, Foster S J, Smythe C, Battaglia G, Thomas J A. Nat. Chem., 2009, 1(8):662.
[81] Tian X H, Gill M R, Canton I, Thomas J A, Battaglia G. ChemBioChem, 2011, 12(4):548.
[82] Gill M R, Derrat H, Smythe C G W, Battaglia G, Thomas J A. ChemBioChem, 2011, 12(6):877.
[83] Zhu B Z, Chevion M. Chem-Biol. Interact., 2000, 129:249.
[84] Zhu B Z, Shechtman S, Chevion M. Chemosphere, 200145:463.
[85] Zhu B Z. Chem. Res. Toxicol., 2001, 14:222.
[86] Barbouti A, Doulias P T, Zhu B Z, Frei B, Galaris D. Free Radic. Res., 2005, 39:125.
[87] Levy S, Shechtman S, Zhu B Z, Stadtman E R, Stadler R, Chevion M. Environ. Toxicol. Chem., 2007, 26:218.
[88] 朱本占(Zhu B Z), 范瑞梅(Fan R M), 盛治国(Sheng Z G).科学通报(Chinese Science Bulletin), 2011, 54:1673.
[89] Sheng Z G, Li Y, Fan R M, Chao X J, Zhu B Z. Toxicol. Appl. Pharmacol., 2013, 266:335.
[90] Fan R M, Zhu B Z, Huang C P, Sheng Z G, Mao L, Li M X. Environ. Toxicol., 2016, 1964.
[91] Zhu B Z, Chao X J, Huang C H, Li Y. Chem. Sci., 2016, 7:4016.
[92] Shao B, Mao L, Qu N, Wang Y F, Gao H Y, Li F, Qin L, Shao J, Huang C H, Xu D, Xie L N, Shen C, Zhou X, Zhu B Z. Free. Radic. Biol. Med., 2017, 104:54.
[93] Hoenger A, McIntosh J R. Curr. Opin. Cell. Biol., 2009, 21(1):89.
[94] Van-Rijt S H, Mukherjee A, Pizarro A M, Sadler P J. J. Med. Chem., 2010, 53(2):840.
[95] Sava G, Zorzet S, Turrin C, Vita F, Soranzo M, Zabucchi G, Cocchietto M, Bergamo A, DiGiovine S, Pezzoni G, Sartor L, Garbisa S. Clin. Cancer. Res., 2003, 9(5):1898.
[96] Meister K, Niesel J, Schatzschneider U, Metzler-Nolte N, Schmidt D A, Havenith M. Angew. Chem. Int. Edit., 2010, 49(19):3310.
[97] Cosgrave L, Devocelle M, Forster R J, Keyes T E. Chem. Commun., 2010, 46(1):103.
[98] O'Connor N A, Stevens N, Samaroo D, Solomon M R, Marti A A, Dyer J, Vishwasrao H, Akins D L, Kandel E R, Turro N J. Chem. Commun., 2009, (19):2640.
[99] Schatzschneider U, Niesel J, Ott I, Gust R, Alborzinia H, Wolfl S. Chem. Med. Chem., 2008, 3(7):1104.
[100] Gianferrara T, Bratsos I, Alessio E. Dalton Trans., 2009, (37):7588.
[101] Fink S L, Cookson B T. Infect. Immun., 2005, 73(4):1907.
[102] Lei W H, Zhou Q X, Jiang G Y Zhang B W, Wang X S. Photoch. Photobio. Sci., 2011, 10(6):887.
[103] Foxon S P, Alamiry M A H, Walker M G, Meijer A J H M, Sazanovich I V, Weinstein J A, Thomas J A. J. Phys. Chem. A, 2009, 113(46):12754.
[104] Sun Y J, Joyce L E, Dickson N M, Turro C. Chem. Commun., 2010, 46(14):2426.
[105] Bolhuis A, Hand L, Marshall J E, Richards A D, Rodger A, Aldrich-Wright J. Eur. J. Pharm. Sci., 2011, 42(4):313.
[106] Tan C P, Lai S S, Wu S H, Hu S, Zhou L J, Chen Y, Wang M X, Zhu Y P, Lian W, Peng W L, Ji L N, Xu A L. J. Med. Chem., 2010, 53(21):7613.
[107] Tan C P, Wu S H, Lai S S, Wang M X, Chen Y, Zhou L J, Zhu Y P, Lian W, Peng W L, Ji L N, Xu A L. Dalton Trans., 2011, 40(34):8611.
[108] Pisani M J, Weber D K, Heimann K, Collins J G, Keene F R. Metallomics., 2010, 2(6):393.
[109] Pisani M J, Fromm P D, Mulyana Y, Clarke R J, Korner H, Heimann K, Collins J G, Keene F R. Chem. Med. Chem., 2011, 6(5):848.
[110] Li F F, Mulyana Y, Feterl M, Warner J M, Collins J G, Keene F R. Dalton Trans., 2011, 40(18):5032.
[111] Mulyana Y, Weber D K, Buck D P, Motti C A, Collins J G, Keene F R. Dalton Trans., 2011, 40(7):1510.
[112] Chen T F, Mei W J, Wong Y S, Liu J, Liu Y N, Xie H S, Zheng W J. MedChemComm, 2010, 1(1):73.
[113] Chen T F, Liu Y A, Zheng W J, Liu J, Wong Y S. Inorg. Chem., 2010, 49(14):6366.
[114] Ernst R J, Song H, Barton J K. J. Am. Chem. Soc., 2009, 131(6):2359.
[115] Aguirre J D, Angeles-Boza A M, Chouai A, Pellois J P, Turro C, Dunbar K R. J. Am. Chem. Soc., 2009, 131(32):11353.
[116] Joyce L E, Aguirre J D, Angeles-Boza A M, Chouai A, Fu P K L, Dunbar K R, Turro C. Inorg. Chem., 2010, 49(12):5371.
[117] Poon C T, Chan P S, Man C, Jiang F L, Wong R N S, Mak N K, Kwong D W J, Tsao S W, Wong W K. J. Inorg. Biochem., 2010, 104(1):62.
[118] Milkevitch M, Storrie H, Brauns E, Brewer K J, Shirley B W. Inorg. Chem., 1997, 36(20):4534.
[119] Van-Der-Schilden K, Garcia F, Kooijman H, Spek A L, Haasnoot J G, Reedijk J. Angew. Chem. Int. Edit., 2004, 43(42):5668.
[120] Petitjean A, Barton J K. J. Am. Chem. Soc., 2004, 126(45):14728.
[121] Ruggi A, Beekman C, Wasserberg D, Subramaniam V, Reinhoudt D N, van Leeuwen F W B, Velders A H. Chem. Eur. J., 2011, 17(2):464.
[122] Elmes R B P, Orange K N, Cloonan S M, Williams D C. J. Am. Chem. Soc., 2011, 133(40):15862.
[123] Murphy C J, Barton J K. Metallobiochemistry Part C, 1993, 226:576.
[124] Puckett C A, Ernst R J, Barton J K. Dalton Trans., 2010, 39(5):1159.
[125] Sun Y J, Collins S N, Joyce L E, Turro C. Inorg. Chem., 2010, 49(9):4257.
[126] Shi H J, Chen Y, Gao F, Yu H J, Li G Y, Chao H, Shi X F, Ji L N. J. Mol. Struct., 2008, 892(1/3):485.
[127] Cook N P, Kilpatrick K, Segatori L, Marti A A. J. Am. Chem. Soc., 2012, 134(51):20776.
[128] Cook N P, Torres V, Jain D, Marti A A. J. Am. Chem. Soc., 2011, 133(29):11121.
[129] Aliyan A, Kirby B, Pennington C, Martí A A. J. Am. Chem. Soc., 2016, 138:8686.
[130] Silva D E S, Cali M P, Pazin W M, Lima E C, Trevisan M T S, Venancio T, Miranda M A, Ito A S, Carlos R M. J. Med. Chem., 2016, 59:9215.
[131] Li G Y, Sun L L, Ji L N, Chao H. Dalton Trans., 2016, 45:13261.
[132] Boynton A N, Marcélis L, Barton J K. J.Am. Chem. Soc., 2016, 138:5020.
[133] Boynton A N, Marcelis L, McConnell A J, Barton J K. Inorg. Chem., 2017, 56:8381.
[134] Sun W, Li S Y, Häupler B, Liu J, Jin S B, Steffen W, Schubert U S, Butt B J, Liang X J, Wu S. Adv. Mater., 2017, 29:1.
[135] Shen J L, Kim H C, Wolfram J, Mu C F, Zhang W, Liu H R, Xie Y, Mai J H, Zhang H, Li Z, Guevara M, Mao Z W, Shen H F. Nano Letters, 2017, 17:2913.
[136] Gao M, Yu F, Lv C J, Choo J, Chen L X. Chem. Soc. Rev., 2017, 46:2237.
PDF
(1517KB)
