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Monitoring the Instant Creation of a New Fluorescent Signal for Evaluation of DNA Conformation Based on Intercalation Complex

  • Ahmet T. Uzumcu
  • Orhan Guney
  • Oguz Okay
Original Article
  • 15 Downloads

Abstract

Here we report the monitoring the instant creation of a new fluorescent signal (FS) aroused from a positively charged water-soluble fluorogenic probe, ethidium bromide (EtBr) in the presence of a radical initiator, ammonium persulfate (APS) and an accelerator, tetraethylmetilendiamine (TEMED) for evaluation of deoxyribonucleic acid (DNA) conformation. The results revealed that the occurred FS (λex = 430 nm; λmax = 525 nm) is a reduced form of EtBr (λex = 480 nm; λmax = 617 nm) and it is completely distinct from hydroethidine (λex = 350 nm; λmax = 430 nm), which is two-electron reduced form of EtBr. It was noticed that EtBr was reduced to a new FS during the polymerization of N, N dimethyacrylamide (DMAA) too, at 25 °C in the presence of APS and TEMED or at 55 °C with only APS, and the rate of formation of FS was increased upon treatment time. The effect of nanoclays such as Laponite XLG® and Laponite XLS®, which provide a protective environment for DNA in nature, were also investigated through the reduction process of EtBr in the absence and presence of a water soluble monomer DMAA. We demonstrated that DNA conformation might be evaluated by monitoring FS effectuated during the reduction of EtBr in the presence of nanoclays having positively and negatively charged surfaces. Protective property of DNA against the formation of reduced product was elucidated by carrying out the polymerization at 55 °C. The results revealed that the monitoring of formation of FS in the presence of radical initiator could lead to elucidate the conformation of DNA upon formation of intercalator complex.

Keywords

EtBr DNA conformation Intercalation complex Nanoclay 

Notes

Acknowledgements

This work was supported by Istanbul Technical University (ITU), BAP 39773. O.O. thanks to Turkish Academy of Sciences (TUBA) for the partial support.

Supplementary material

10895_2018_2294_MOESM1_ESM.docx (851 kb)
ESM 1 (DOCX 850 kb)

References

  1. 1.
    Lavis LD, Raines RT (2008) Bright ideas for chemical biology. ACS Chem Biol 3:142–155CrossRefGoogle Scholar
  2. 2.
    Terai T, Nagano T (2008) Fluorescent probes for bioimaging applications. Curr Opin Chem Biol 12:515–521CrossRefGoogle Scholar
  3. 3.
    Wysocki LM, Lavis LD (2011) Advances in the chemistry of small molecule fluorescent probes. Curr Opin Chem Biol 15:752–759CrossRefGoogle Scholar
  4. 4.
    Chib R, Raut S, Sabnis S, Singhal P, Gryczynski Z, Gryczynski I (2014) Associated anisotropy decays of ethidium bromide interacting with DNA. Methods Appl Fluoresc 2CrossRefGoogle Scholar
  5. 5.
    Mukherjee A, Singh B (2017) Binding interaction of phatinaceutical drug captopril with calf thymus DNA: a multispectroscopic and molecular docking study. J Lumin 190:319–327CrossRefGoogle Scholar
  6. 6.
    Liu BM, Bai CL, Zhang J, Liu Y, Dong BY, Zhang YT, Liu B (2015) In vitro study on the interaction of 4,4-dimethylcurcumin with calf thymus DNA. J Lumin 166:48–53CrossRefGoogle Scholar
  7. 7.
    Roy S, Saxena SK, Mishra S, Yogi P, Sagdeo PR, Kumar R (2018) Spectroscopic evidence of phosphorous heterocycle-DNA interaction and its verification by docking approach. J Fluoresc 28:373–380CrossRefGoogle Scholar
  8. 8.
    Cosa G, Focsaneanu KS, McLean JRN, McNamee JP, Scaiano JC (2001) Photophysical properties of fluorescent DNA-dyes bound to single- and double-stranded DNA in aqueous buffered solution. Photochem Photobiol 73:585–599CrossRefGoogle Scholar
  9. 9.
    Sun YT, Peng TT, Zhao L, Jiang DY, Cui YC (2014) Studies of interaction between two alkaloids and double helix DNA. J Lumin 156:108–115CrossRefGoogle Scholar
  10. 10.
    Paramanik B, Bhattacharyya S, Patra A (2013) Steady state and time resolved spectroscopic study of QD-DNA interaction. J Lumin 134:401–407CrossRefGoogle Scholar
  11. 11.
    Yuvaraj M, Aruna P, Koteeswaran D, Tamilkumar P, Ganesan S (2015) Rapid fluorescence spectroscopic characterization of salivary DNA of normal subjects and OSCC patients using ethidium bromide. J Fluoresc 25:79–85CrossRefGoogle Scholar
  12. 12.
    Geng SQ, Wu Q, Shi L, Cui FL (2013) Spectroscopic study one thiosemicarbazone derivative with ctDNA using ethidium bromide as a fluorescence probe. Int J Biol Macromol 60:288–294CrossRefGoogle Scholar
  13. 13.
    Zhou XY, Zhang GW, Wang LH (2014) Binding of 8-methoxypsoralen to DNA in vitro: monitoring by spectroscopic and chemometrics approaches. J Lumin 154:116–123CrossRefGoogle Scholar
  14. 14.
    Agarwal S, Jangir DK, Mehrotra R (2013) Spectroscopic studies of the effects of anticancer drug mitoxantrone interaction with calf-thymus DNA. J Photochem Photobiol B Biol 120:177–182CrossRefGoogle Scholar
  15. 15.
    Rafique B, Khalid AM, Akhtar K, Jabbar A (2013) Interaction of anticancer drug methotrexate with DNA analyzed by electrochemical and spectroscopic methods. Biosens Bioelectron 44:21–26CrossRefGoogle Scholar
  16. 16.
    Temerk Y, Ibrahim M, Ibrahim H, Kotb M (2015) Interactions of an anticancer drug Formestane with single and double stranded DNA at physiological conditions. J Photochem Photobiol B Biol 149:27–36CrossRefGoogle Scholar
  17. 17.
    Xia KX, Zhang GW, Li S, Gong DM (2017) Groove binding of vanillin and ethyl vanillin to calf Thymus DNA. J Fluoresc 27:1815–1828CrossRefGoogle Scholar
  18. 18.
    Benov L, Sztejnberg L, Fridovich I (1998) Critical evaluation of the use of hydroethidine as a measure of superoxide anion radical. Free Radic Biol Med 25:826–831CrossRefGoogle Scholar
  19. 19.
    Budd SL, Castilho RF, Nicholls DG (1997) Mitochondrial membrane potential and hydroethidine-monitored superoxide generation in cultured cerebellar granule cells. FEBS Lett 415:21–24CrossRefGoogle Scholar
  20. 20.
    Zhao H, Kalivendi S, Zhang H, Joseph J, Nithipatikom K, Vásquez-Vivar J, Kalyanaraman B (2003) Superoxide reacts with hydroethidine but forms a fluorescent product that is distinctly different from ethidium: potential implications in intracellular fluorescence detection of superoxide. Free Radic Biol Med 34:1359–1368CrossRefGoogle Scholar
  21. 21.
    Zielonka J, Kalyanaraman B (2010) Hydroethidine- and MitoSOX-derived red fluorescence is not a reliable indicator of intracellular superoxide formation: another inconvenient truth. Free Radic Biol Med 48:983–1001CrossRefGoogle Scholar
  22. 22.
    Kalyanaraman B, Dranka BP, Hardy M, Michalski R, Zielonka J (2014) HPLC-based monitoring of products formed from hydroethidine-based fluorogenic probes - the ultimate approach for intra- and extracellular superoxide detection. Biochim Biophys Acta-General Subjects 1840:739–744CrossRefGoogle Scholar
  23. 23.
    Maghzal GJ, Cergol KM, Shengule SR, Suarna C, Newington D, Kettle AJ, Payne RJ, Stocker R (2014) Assessment of myeloperoxidase activity by the conversion of hydroethidine to 2-chloroethidium. J Biol Chem 289:5580–5595CrossRefGoogle Scholar
  24. 24.
    Michalski R, Michalowski B, Sikora A, Zielonka J, Kalyanaraman B (2014) On the use of fluorescence lifetime imaging and dihydroethidium to detect superoxide in intact animals and ex vivo tissues: a reassessment. Free Radic Biol Med 67:278–284CrossRefGoogle Scholar
  25. 25.
    Thomas G, Roques B (1972) Proton magnetic-resonance studies of ethidium bromide and its sodium-borohydride reduced derivative. Febs Lett 26:169–175CrossRefGoogle Scholar
  26. 26.
    Phukan S, Mitra S (2012) Fluorescence behavior of ethidium bromide in homogeneous solvents and in presence of bile acid hosts. J Photochem Photobiol A Chem 244:9–17CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Departments of Chemistry and Polymer Science & TechnologyIstanbul Technical UniversityIstanbulTurkey

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