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Photophysical and calorimetric investigation on the structural reorganization of poly(A) by phenothiazinium dyes azure A and azure B

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Abstract

Poly(A) has significant relevance to mRNA stability, protein synthesis and cancer biology. The ability of two phenothiazinium dyes azure A (AA) and azure B (AB) to bind single-stranded poly(A) was studied by spectroscopic and calorimetric techniques. Strong binding of the dyes and the higher affinity of AA over AB were ascertained from absorbance and fluorescence experiments. Significant perturbation of the circular dichroism spectrum of poly(A) in the presence of these molecules with formation of induced CD bands in the 300–700 nm region was observed. Strong emission polarization of the bound dyes and strong energy transfer from the adenine base pairs of poly(A) suggested intercalative binding to poly(A). Intercalative binding was confirmed from fluorescence quenching experiments and was predominantly entropy driven as evidenced from isothermal titration calorimetry data. The negative values of heat capacity indicated involvement of hydrophobic forces and enthalpy–entropy compensation suggested noncovalent interactions in the complexation for both the dyes. Poly(A) formed a self-assembled structure on the binding of both the dyes that was more favored under higher salt conditions. New insights in terms of spectroscopic and thermodynamic aspects into the self-structure formation of poly(A) by two new phenothiazinium dyes that may lead to structural and functional damage of mRNA are revealed from these studies.

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References

  1. S. L. Topalian, S. Kaneko, M. I. Gonzales, G. Bond, Y. Ward, J. Manley, Identification and functional characterization of neo-poly(A) polymerase, an RNA processing enzyme overexpressed in human tumors, Mol. Cell. Biol., 2001, 21, 5614–5623.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. H. Xi, D. Gray, S. Kumar, D. P. Arya, Molecular recognition of single-stranded RNA: neomycin binding to poly(A), FEBS Lett., 2009, 583, 2269–2275.

    Article  CAS  PubMed  Google Scholar 

  3. P. Giri, M. Hossain, G. S. Kumar, RNA specific molecules: cytotoxic plant alkaloid palmatine binds strongly to poly(A), Bioorg. Med. Chem. Lett., 2006, 16, 2364–2368.

    Article  CAS  PubMed  Google Scholar 

  4. M. Hossain, A. Kabir, G. S. Kumar, Binding of the phenothiazinium dye methylene blue with single stranded polyriboadenylic acid, Dyes Pigm., 2012, 92, 1376–1383.

    Article  CAS  Google Scholar 

  5. P. Giri, G. S. Kumar, Self-structure induction in single stranded poly(A) by small molecules: studies on DNA intercalators, partial intercalators and groove binding molecules, Arch. Biochem. Biophys., 2008, 474, 183–192.

    Article  CAS  PubMed  Google Scholar 

  6. F. Xing, G. Song, J. Ren, J. B. Chaires, X. Qu, Molecular recognition of nucleic acids: coralyne binds strongly to poly(A), FEBS Lett., 2005, 579, 5035–5039.

    Article  CAS  PubMed  Google Scholar 

  7. P. Giri, G. S. Kumar, Specific binding and self-structure induction to poly(A) by the cytotoxic plant alkaloid sanguinarine, Biochim. Biophys. Acta, 2007, 1770, 1419–1426.

    Article  CAS  PubMed  Google Scholar 

  8. P. Giri, G. S. Kumar, Molecular recognition of poly(A) targeting by protoberberine alkaloids: in vitro biophysical studies and biological perspectives, Mol. BioSyst., 2010, 6, 81–88.

    Article  CAS  PubMed  Google Scholar 

  9. M. M. Islam, A. Basu, G. S. Kumar, Binding of 9-O-(ω-amino) alkyl ether analogues of the plant alkaloid berberine to poly(A): insights into self-structure induction, Med. Chem. Commun., 2011, 2, 631–637.

    Article  CAS  Google Scholar 

  10. A. Basu, P. Jaisankar, G. S. Kumar, 9-O-N-aryl/arylalkyl amino carbonyl methyl substituted berberine analogues induce self-structure in polyadenylic acid, RSC Adv., 2012, 2, 7714–7723.

    Article  CAS  Google Scholar 

  11. S. Taniguchi, N. Suzuki, M. Masuda, S. Hisanaga, T. Iwatsubo, M. Goedert, M. Hasegawa, Inhibition of heparin induced tau filament formation by phenothiazines, polyphenols, and porphyrins, J. Biol. Chem., 2005, 280, 7614–7623.

    Article  CAS  PubMed  Google Scholar 

  12. K. J. Mellish, R. D. Cox, D. I. Vernon, J. Griffiths, S. B. Brown, In vitro photodynamic activity of a series of methylene blue analogues, Photochem. Photobiol., 2002, 75, 392–397.

    Article  CAS  PubMed  Google Scholar 

  13. L. M. Moreira, J. P. Lyon, A. P. Romani, D. Severino, M. R. Rodrigues and H. P. M. de Oliveira, Phenotiazinium dyes as photosensitizers (PS) in photodynamic therapy (PDT): spectroscopic properties and photochemical mechanisms (Chapter 14), in Advanced aspects of spectroscopy, pp. 393–422.

  14. R. F. Donnelly, P. A. McCarron, M. M. Tunney, A. D. Woolfson, Potential of photodynamic therapy in treatment of fungal infections of the mouth. Design and characterization of a mucoadhesive patch containing toluidine blue O, J. Photochem. Photobiol., B, 2007, 86, 59–69.

    Article  CAS  Google Scholar 

  15. L. M. Moreira, J. P. Lyon, S. M. S. Tursi, I. Trajano, M. P. Felipe, M. S. Costa, M. R. Rodrigues, L. Codognoto, H. P. M. de Oliveira, Azure dyes as new photosensitizer prototypes to application in photodynamic therapy against Candida spp., Spectroscopy, 2010, 24, 621–628.

    Article  CAS  Google Scholar 

  16. M. Wainwright, M. N. Byrne, M. A. Gattrell, Phenothiazinium-based photobactericidal materials, J. Photochem. Photobiol., B, 2006, 84, 227–230.

    Article  CAS  Google Scholar 

  17. E. L. Kean, Rapid, sensitive spectrophotometric method for quantitative determination of sulfatides, J. Lipid Res., 1968, 9, 319–327.

    Article  CAS  PubMed  Google Scholar 

  18. G. R. Webb, I. A. Macdonald, C. N. Williams, Ion pair extraction technique with azure A, for differentiating biles of normal subjects and patients with liver disease from patients with Crohn’s disease with small bowel involvement, Clin. Chem., 1977, 23, 460–463.

    Article  CAS  PubMed  Google Scholar 

  19. A. K. Baruia, R. Sharma, Y. S. Rajput, Detection of non-dairy fat in milk based on quantitative assay of anionic detergent using azure A dye, Int. Dairy J., 2012, 24, 44–47.

    Article  CAS  Google Scholar 

  20. Y. E. Zeng, H. S. Zhang and Z. H. Chen, Handbook of Organic Reagents, Chemical Industry Press, Beijing, 1989,vol. 4, p. 793.

    Google Scholar 

  21. F. Culo, D. Sabolović, L. Somogyi, M. Marusić, N. Berbiguier, L. Galey, Anti-tumoral and anti-inflammatory effects of biological stains, Agents Actions, 1991, 34, 424–428.

    Article  CAS  PubMed  Google Scholar 

  22. R. H. Schirmer, H. Adler, M. Pickhardt, E. Mandelkow, Lest we forget you—methylene blue..., Neurobiol. Aging, 2011, 32, 2325.e7–16.

    Article  CAS  Google Scholar 

  23. B. Narayana, K. Sunil, Facile and sensitive spectrophotometric method for the determination of vanadium, Eur. J. Anal. Chem., 2009, 4, 141–151.

    Google Scholar 

  24. M. Mathew, B. Narayana, An easy spectrophotometric determination of selenium using azure B as a chromogenic reagent, Indian J. Chem. Technol., 2006, 13, 455–458.

    CAS  Google Scholar 

  25. A. Petzer, B. H. Harvey, G. Wegener, J. P. Petzer, Azure B, a metabolite of methylene blue, is a high-potency, reversible inhibitor of monoamine oxidase, Toxicol. Appl. Pharmacol., 2012, 258, 403–409.

    Article  CAS  PubMed  Google Scholar 

  26. P. Paul, G. S. Kumar, Spectroscopic studies on the binding interaction of phenothiazinium dyes toluidine blue O, azure A and azure B to DNA, Spectrochim. Acta, Part A, 2013, 107, 303–310.

    Article  CAS  Google Scholar 

  27. J. D. McGhee, P. H. von Hippel, Theoretical aspects of DNA-protein interactions: cooperative and noncooperative binding of large ligands to a one dimensional homogeneous lattice, J. Mol. Biol., 1974, 86, 469–489.

    Article  CAS  PubMed  Google Scholar 

  28. A. Das, K. Bhadra, G. S. Kumar, Targeting RNA by small molecules: comparative structural and thermodynamic aspects of aristololactam-β-D-glucoside and daunomycin binding to tRNAphe, PLoS ONE, 2011, 6, 8 e23186.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. P. Giri, G. S. Kumar, Spectroscopic and calorimetric studies on the binding of the phototoxic and cytotoxic plant alkaloid sanguinarine with double helical poly(A), J. Photochem. Photobiol., A, 2008, 194, 111–121.

    Article  CAS  Google Scholar 

  30. R. Lyng, T. Hard, B. Norden, Induced CD of DNA intercalators: electric dipole allowed transitions, Biopolymers, 1987, 26, 1327–1345.

    Article  CAS  PubMed  Google Scholar 

  31. P. Centikol, N. V. Hud, Molecular recognition of poly(A) by small ligands: an alternative method of analysis reveals nanomolar, cooperative and shape-selective binding, Nucleic Acids Res., 2009, 37, 611–621.

    Article  CAS  Google Scholar 

  32. P. Giri, G. S. Kumar, Molecular aspects of small molecules-poly(A) interaction: an approach to RNA based drug design, Curr. Med. Chem., 2009, 16, 965–987.

    Article  CAS  PubMed  Google Scholar 

  33. M. M. Islam, S. R. Chowdhury, G. S. Kumar, Spectroscopic and calorimetric studies on the binding of alkaloids berberine, palmatine and coralyne to double stranded RNA polynucleotides, J. Phys. Chem. B, 2009, 113, 1210–1224.

    Article  CAS  PubMed  Google Scholar 

  34. P. Paul, M. Hossain, G. S. Kumar, Calorimetric and thermal analysis studies on the binding of phenothiazinium dye thionine with DNA polynucleotides, J. Chem. Thermodyn., 2011, 43, 1036–1043.

    Article  CAS  Google Scholar 

  35. M. Hossain, P. Giri, G. S. Kumar, DNA intercalation by quinacrine and methylene blue: a comparative binding and thermodynamic characterization study, DNA Cell Biol., 2008, 27, 81–90.

    Article  CAS  PubMed  Google Scholar 

  36. I. Saha, M. Hossain, G. S. Kumar, Sequence selective binding of phenazinium dyes, phenosafranin and safranin O to guanine-cytosine deoxyribopolynucleotides: spectroscopic and thermodynamic studies, J. Phys. Chem. B, 2010, 114, 15278–15287.

    Article  CAS  PubMed  Google Scholar 

  37. A. Basu, P. Jaisankar, G. S. Kumar, Binding of the 9-O-N-aryl/arylalkyl amino carbonyl methyl substituted berberine analogs to tRNAphe, PLoS ONE, 2013, 8, 3 e58279.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. M. Hossain, A. Kabir, G. S. Kumar, Binding of the anticancer alkaloid sanguinarine with tRNAphe: spectroscopic and calorimetric studies, J. Biomol. Struct. Dyn., 2012, 30, 223–234.

    Article  CAS  PubMed  Google Scholar 

  39. J. Gómez, E. Freire, Thermodynamic mapping of the inhibitor site of the aspartic protease endothiapepsin, J. Mol. Biol., 1995, 252, 337–350.

    Article  PubMed  Google Scholar 

  40. J. Ren, T. C. Jenkins, J. B. Chaires, Energetics of DNA intercalation reactions, Biochemistry, 2000, 39, 8439–8447.

    Article  CAS  PubMed  Google Scholar 

  41. K. L. Buchmueller, S. L. Bailey, D. A. Matthews, Z. T. Taherbhai, J. K. Register, Z. S. Davis, C. D. Bruce, C. O’Hare, J. A. Hartley, M. Lee, Physical and structural basis for the strong interactions of the–ImPy-central pairing motif in the polyamide f-ImPyIm, Biochemistry, 2006, 45, 13551–13565.

    Article  CAS  PubMed  Google Scholar 

  42. F. V. Murphy 4th, M. E. Churchill, Nonsequence-specific DNA recognition: a structural perspective, Structure, 2000, 8, R83–R89.

    Article  CAS  PubMed  Google Scholar 

  43. J. H. Ha, R. S. Spolar, M. T. Record Jr., Role of the hydrophobic effect in stability of site specific protein-DNA complexes, J. Mol. Biol., 1989, 209, 801–816.

    Article  CAS  PubMed  Google Scholar 

  44. K. M. Guthrie, A. D. C. Parenty, L. V. Smith, L. Cronin, A. Cooper, Microcalorimetry of interaction of dihydro-imidazo-phenanthridinium (DIP)-based compounds with duplex DNA, Biophys. Chem., 2007, 126, 117–123.

    Article  CAS  PubMed  Google Scholar 

  45. M. M. Islam, P. Pandya, S. Kumar, G. S. Kumar, RNA targeting through binding of small molecules: studies on t-RNA binding by the cytotoxic protoberberine alkaloid coralyne, Mol. BioSyst., 2009, 5, 244–254.

    Article  CAS  PubMed  Google Scholar 

  46. L. Jen-Jacobson, L. E. Engler, L. A. Jacobson, Structural and thermodynamic strategies for site-specific DNA binding proteins, Structure, 2000, 8, 1015–1023.

    Article  CAS  PubMed  Google Scholar 

  47. E. V. Starikov, B. Norden, Enthalpy–entropy compensation: a phantom or something useful?, J. Phys. Chem. B, 2007, 111, 14431–14435.

    Article  CAS  PubMed  Google Scholar 

  48. C. Ciatto, M. L. D’Amico, G. Natile, F. Secco, M. Venturini, Intercalation of proflavine and a platinum derivative of proflavine into double-helical poly(A), Biophys. J., 1999, 77, 2717–2724.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. R. Nandi, D. Debnath, M. Maiti, Interactions of berberine with poly(A) and tRNA, Biochim. Biophys. Acta, 1990, 1049, 339–342.

    Article  CAS  PubMed  Google Scholar 

  50. A. Chakraborty, M. Ali, S. K. Saha, Molecular interaction of organic dyes in bulk and confined media, Spectrochim. Acta Part A, 2010, 75, 1577–1583.

    Article  CAS  Google Scholar 

  51. R. Sinha, M. M. Islam, K. Bhadra, G. S. Kumar, A. Banerjee, M. Maiti, The binding of DNA intercalating and non-intercalating compounds to A-form and protonated form of poly(rC).poly(rG): spectroscopic and viscometric study, Bioorg. Med. Chem., 2006, 14, 800–814.

    Article  CAS  PubMed  Google Scholar 

  52. P. Job, Formation and stability of inorganic complexes in solution, Ann. Chim., 1928, 9, 113–203.

    CAS  Google Scholar 

  53. C. Y. Huang, S. P. Colowick and N. O. Kaplan, Methods Enzymol., Academic Press, New York, 1982, vol. 87, pp. 509–525.

    Article  CAS  PubMed  Google Scholar 

  54. Z. D. Hill, M. Patrick, Novel approach to Job’s method: an undergraduate experiment, J. Chem. Educ., 1986, 63, 162–167.

    Article  CAS  Google Scholar 

  55. A. Larsson, C. Carlsson, M. Jonsson, B. Albinsson, Characterization of the binding of the fluorescent dyes YO and YOYO to DNA by polarized light spectroscopy, J. Am. Chem. Soc., 1994, 116, 8459–8465.

    Article  CAS  Google Scholar 

  56. C. V. Kumar, E. H. Asuncion, Sequence dependent energy transfer from DNA to a simple aromatic chromophore, J. Chem. Soc., Chem. Commun., 1992, 6, 470–472.

    Article  Google Scholar 

  57. N. K. Modukuru, K. J. Snow, B. S. Perrin Jr., A. Bhambhani, M. Duff, C. V. Kumar, Tuning the DNA binding modes of an anthracene derivative with salt, J. Photochem. Photobiol., A, 2006, 177, 43–54.

    Article  CAS  Google Scholar 

  58. S. Das, G. S. Kumar, Molecular aspects on the interaction of phenosafranine to deoxyribonucleic acid: model for intercalative drug-DNA binding, J. Mol. Struct., 2008, 872, 56–63.

    Article  CAS  Google Scholar 

  59. M. M. Islam, R. Sinha, G. S. Kumar, RNA binding small molecules: studies on t-RNA binding by cytotoxic plant alkaloids berberine, palmatine and the comparison to ethidium, Biophys. Chem., 2007, 125, 508–520.

    Article  CAS  PubMed  Google Scholar 

  60. M. M. Islam, P. Pandya, S. R. Chowdhury, S. Kumar, G. S. Kumar, Binding of DNA-binding alkaloids berberine and palmatine to tRNA and comparison to ethidium: spectroscopic and molecular modeling studies, J. Mol. Struct., 2008, 891, 498–507.

    Article  CAS  Google Scholar 

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Correspondence to Gopinatha Suresh Kumar.

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Paul, P., Kumar, G.S. Photophysical and calorimetric investigation on the structural reorganization of poly(A) by phenothiazinium dyes azure A and azure B. Photochem Photobiol Sci 13, 1192–1202 (2014). https://doi.org/10.1039/c4pp00085d

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