Advertisement

Science China Chemistry

, Volume 61, Issue 12, pp 1630–1636 | Cite as

A dsDNA-lighted fluorophore for monitoring protein-ligand interaction through binding-mediated DNA protection

  • Zhihe QingEmail author
  • Lixuan Zhu
  • Lina Hou
  • Zhen Zou
  • Sheng Yang
  • Ronghua YangEmail author
Articles
  • 23 Downloads

Abstract

Because of their important roles in cellular functions, life activities, drug screening, and disease treatment, the development of efficient methods for monitoring protein-ligand interaction is essential. In this study, inspired by our previous studies on DNA conformation-selective fluorescent indicators, we developed a new sensing platform for monitoring protein-ligand interaction and detecting protein activity based on binding-mediated DNA protection and the dsDNA-lighted fluorophore, ethyl-4-[3,6-bis (1-methyl-4-vinylpyridium iodine)-9H-carbazol-9-yl)] butanoate (EBCB). The ligand was purposefully linked to the 3ʹ-terminal of a hairpin DNA probe to selectively bind with the target protein and protect the DNA from cleavage by exonuclease III. By virtue of EBCB’s outstanding capacity to discriminate DNA conformation, the protein-ligand interaction could be effectively monitored through a fluorescence change in EBCB. A high fluorescence signal was detected when the hairpin DNA was protected in the presence of the target protein, whereas a much lower signal was observed in the presence of nontarget proteins. Our results demonstrated that the proposed strategy had high potential, such as high selectivity and relatively high sensitivity, for monitoring protein-ligand interaction and detecting protein activity. We believe these results will pave the way for applying dsDNA-lighted fluorophore EBCB as an effective signal transducer for DNA conformation transformation-mediated biochemical sensing.

Keywords

dsDNA-lighted fluorophore protein-ligand interaction DNA protection fluorescence detection 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (21605008, 21735001, 21575018, 21505006) and the Hunan Provincial Natural Science Foundation (2016JJ3001).

Supplementary material

11426_2018_9349_MOESM1_ESM.doc (2.6 mb)
A dsDNA-lighted fluorophore for monitoring protein–ligand interaction through binding-mediated DNA protection

References

  1. 1.
    Heal WP, Dang THT, Tate EW. Chem Soc Rev, 2011, 40: 246–257CrossRefGoogle Scholar
  2. 2.
    Bai Y, Luo Q, Liu J. Chem Soc Rev, 2016, 45: 2756–2767CrossRefGoogle Scholar
  3. 3.
    Li D, Lu G, Lei C, Wang Z, Li L, Nie Z, Huang Y, Yao S. Sci China Chem, 2016, 59: 809–815CrossRefGoogle Scholar
  4. 4.
    Jones S, Thornton JM. Proc Natl Acad Sci USA, 1996, 93: 13–20CrossRefGoogle Scholar
  5. 5.
    Inohara N, Chamaillard M, McDonald C, Nuñez G. Annu Rev Biochem, 2005, 74: 355–383CrossRefGoogle Scholar
  6. 6.
    Stockwell BR. Nature, 2004, 432: 846–854CrossRefGoogle Scholar
  7. 7.
    Zhou DM, Wu YD, Liu P, Bai HT, Tang LJ, Yu RQ, Jiang JH. Sci China Chem, 2011, 54: 1277–1283CrossRefGoogle Scholar
  8. 8.
    Michnick SW, Ear PH, Manderson EN, Remy I, Stefan E. Nat Rev Drug Discov, 2007, 6: 569–582CrossRefGoogle Scholar
  9. 9.
    Zhang T, Wei T, Han Y, Ma H, Samieegohar M, Chen PW, Lian I, Lo YH. AC. Cent Sci, 2016, 2: 834–842CrossRefGoogle Scholar
  10. 10.
    Freyer MW, Lewis EA. Methods Cell Biol, 2008, 84: 79–113CrossRefGoogle Scholar
  11. 11.
    Cooper MA. Nat Rev Drug Discov, 2002, 1: 515–528CrossRefGoogle Scholar
  12. 12.
    Wear MA, Patterson A, Malone K, Dunsmore C, Turner NJ, Walkinshaw MD. Anal Biochem, 2005, 345: 214–226CrossRefGoogle Scholar
  13. 13.
    Lee CK, Wang YM, Huang LS, Lin S. Micron, 2007, 38: 446–461CrossRefGoogle Scholar
  14. 14.
    Star A, Gabriel JCP, Bradley K, Grüner G. Nano Lett, 2003, 3: 459–463CrossRefGoogle Scholar
  15. 15.
    Østergaard J, Jensen H, Holm R. Sep Sci, 2009, 32: 1712–1721CrossRefGoogle Scholar
  16. 16.
    Zhu L, Qing Z, Hou L, Yang S, Zou Z, Cao Z, Yang R. ACS Sens, 2017, 2: 1198–1204CrossRefGoogle Scholar
  17. 17.
    Qing Z, Zhu L, Li X, Yang S, Zou Z, Guo J, Cao Z, Yang R. Environ Sci Technol, 2017, 51: 11884–11890CrossRefGoogle Scholar
  18. 18.
    Qing Z, Hou L, Yang L, Zhu L, Yang S, Zheng J, Yang R. Anal Chem, 2016, 88: 9759–9765CrossRefGoogle Scholar
  19. 19.
    Cash KJ, Ricci F, Plaxco KW. Am Chem Soc, 2009, 131: 6955–6957CrossRefGoogle Scholar
  20. 20.
    Wu Z, Wang H, Guo M, Tang LJ, Yu RQ, Jiang JH. Anal Chem, 2011, 83: 3104–3111CrossRefGoogle Scholar
  21. 21.
    Mohamma. Danesh N, Ramezani M, Sarreshtehdar Emrani A, Abnous K, Taghdisi SM. Biosens Bioelectron, 2016, 75: 123–128CrossRefGoogle Scholar
  22. 22.
    Du Y, Dong S. Anal Chem, 2016, 89: 189–215CrossRefGoogle Scholar
  23. 23.
    Tang Q, Wang N, Zhou F, Deng T, Zhang S, Li J, Yang R, Zhong W, Tan W. Chem Commun, 2015, 51: 16810–16812CrossRefGoogle Scholar
  24. 24.
    Perry LJ, Wetzel R. Science, 1984, 226: 555–557CrossRefGoogle Scholar
  25. 25.
    Bota DA, Davies KJA. Nat Cell Biol, 2002, 4: 674–680CrossRefGoogle Scholar
  26. 26.
    Lesniak A, Campbell A, Monopoli MP, Lynch I, Salvati A, Dawson KA. Biomaterials, 2010, 31: 9511–9518CrossRefGoogle Scholar
  27. 27.
    Lingwood D, Simons K. Science, 2010, 327: 46–50CrossRefGoogle Scholar
  28. 28.
    Lee CC, Avalos AM, Ploegh HL. Nat Rev Immunol, 2012, 12: 168–179CrossRefGoogle Scholar
  29. 29.
    Czabotar PE, Lessene G, Strasser A, Adams JM. Nat Rev Mol Cell Biol, 2014, 15: 49–63CrossRefGoogle Scholar
  30. 30.
    Xu Y, Shen J, Luo X, Shen X, Chen K, Jiang H. Sci China Ser BChem, 2004, 47: 355–366CrossRefGoogle Scholar
  31. 31.
    Wang M, Li F, Jiang B, Xu J, Li S, Xiang Y, Yuan R. Sci China Chem, 2016, 59: 770–775CrossRefGoogle Scholar
  32. 32.
    Wang B, Feng W, Chai Z, Zhao Y. Sci China Chem, 2015, 58: 768–779CrossRefGoogle Scholar
  33. 33.
    Shevelev IV, Hübscher U. Nat Rev Mol Cell Biol, 2002, 3: 364–376CrossRefGoogle Scholar
  34. 34.
    Hu P, Zhu C, Jin L, Dong S. Biosens Bioelectron, 2012, 34: 83–87CrossRefGoogle Scholar
  35. 35.
    Rothschild LJ, Mancinelli RL. Nature, 2001, 409: 1092–1101CrossRefGoogle Scholar
  36. 36.
    Nagajyoti PC, Lee KD, Sreekanth TVM. Environ Chem Lett, 2010, 8: 199–216CrossRefGoogle Scholar

Copyright information

© Science China Press and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Hunan Provincial Key Laboratory of Materials Protection for Electric Power and Transportation, Hunan Provincial Engineering Research Center for Food Processing of Aquatic Biotic Resources, School of Chemistry and Biological EngineeringChangsha University of Science and TechnologyChangshaChina

Personalised recommendations