Spectroscopic study of pheophorbide-a methyl ether binding to synthetic polynucleotides and DNA

  • О. А. Ryazanova B. Verkin Institute for Low Temperature Physics and Engineering of NAS of Ukraine
  • I. М. Voloshin B. Verkin Institute for Low Temperature Physics and Engineering of NAS of Ukraine
  • L. V. Dubey Institute of Molecular Biology and Genetics of NAS of Ukraine
  • V. N. Zozulya B. Verkin Institute for Low Temperature Physics and Engineering of NAS of Ukraine
Keywords: pheophorbide-a, polynucleotides, DNA, intercalation, fluorescence, absorption

Abstract

Binding of pheophorbide-a methyl ether (Mepheo-a) to synthetic double-stranded poly(A)poly(U),
poly(G)poly(C) polynucleotides, four-stranded poly(G), and native DNA was studied by methods of
absorption and polarized fluorescent spectroscopy. Measurements were carried out in aqueous buffered
solutions pH6.9 of low ionic strength (2 мM Na+) in a wide range of phosphate-to-dye molar ratios (P/D).
Absorption and fluorescent characteristics of complexes formed by the dye with the biopolymers were
determined. It is established that binding of neutral Mepheo-a to four-stranded poly(G) is accompanied by
significant spectral transformations, namely, by dye absorption hypochromic, the bathochromic shift of Soret
absorption band (~26 nm), the bathochromic shift of fluorescence band maximum (~9 nm) and 50-fold increase of dye fluorescence intensity. Unlike that, Mepheo-a binding to double-stranded poly(A)poly(U),
poly(G)poly(C) polynucleotides, and to calf thymus DNA is accompanied by insignificant spectral shifts of
absorption and fluorescence bands, as well as by no more than 4-fold rise of fluorescence intensity. Observed
substantial spectral changes and high value of fluorescence polarization degree (0.26) evidence the intercalation of dye chromophore to four-stranded polynucleotide structure. An insignificant increase of dye fluorescence polarization degree (0.12) and weaker spectral transformations point to another binding mode, namely, incorporation of MePheo-а to a groove of the double helix, presumably, in the dimeric form. Since binding of MeРheo-a to four-stranded poly(G) polynucleotide induces substantial changes in fluorescent characteristics of the dye, this derivative is promising as a fluorescent probe for monitoring the G-quadruplex structure.

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Author Biographies

О. А. Ryazanova, B. Verkin Institute for Low Temperature Physics and Engineering of NAS of Ukraine

47 Lenin ave., 61103, Kharkiv,
ryazanova@ilt.kharkov.ua

I. М. Voloshin, B. Verkin Institute for Low Temperature Physics and Engineering of NAS of Ukraine

47 Lenin ave., 61103, Kharkiv
ryazanova@ilt.kharkov.ua

L. V. Dubey, Institute of Molecular Biology and Genetics of NAS of Ukraine

150 Zabolotnogo str., 03143, Kiev

V. N. Zozulya, B. Verkin Institute for Low Temperature Physics and Engineering of NAS of Ukraine

47 Lenin ave., 61103, Kharkiv
ryazanova@ilt.kharkov.ua

References

Owens J.W., Smith R., Robinson R. and Robins M. // Inorg. Chim. Acta.– 1998.– v. 279.– p. 226–231.

Dougherty T.Jr., Gomer C.J., Henderson B.W., et al. // J. Natl. Cancer Inst.– 1998.– v. 90.– p. 889– 905.

Ricchelli F. // J. Photochem. Photobiol. B: Biol. – 1995.– v. 29.– p. 109–111.

Willstatter R. and Stoll A. Untersuchungen über Chlorophyll. Methoden und Ergebnisse. – Berlin: Springer, 1913.

Röder B. // J. Photochem. Photobiol. B: Biol. – 1990. – v.5. – p. 519-521.

Röder B. // Lasers Med. Sci.– 1990.– v.5.– p.99-106.

Roeder B. Pheophorbides. – Photodynamic Tumor Therapy: 2nd and 3rd Generation Photosensitizers. (J.G. Moser, Ed.) .– Amsterdam: Harwood Academic Publishers. – 1998.– p. 35-42.

Hajri A., Wack S. et al. // Photochem. Photobiol.– 2002.– v.75.– p.140-148.

MacDonald I.J. and Dougherty T.Jr. // J. Porphyrins Phthalocyanines.– 2001.- v. 5.– p.105-129.

Krasnovsky A. Jr. et al. // J. Photochem Photobiol. B: Biol.– 1990.– v.5.– p. 245-254.

Roeder B. Photobiophysical parameters. – Photodynamic Tumor Therapy: 2nd and 3rd Generation Photosensitizers. (J.G. Moser, Ed.). – Amsterdam: Harwood Academic Publishers. – 1998.– p. 9-19.

Tanielian C., Kobayashi M., Wolf C. // J. Biomed. Opt. – 2001. – v. 6. – p. 252-256.

Eichwurzel I., Stiel H., Roeder B. // J. Photochem. Photobiol. B.: Biol. – 2000.– v.54.– p.194-200.

Kobayashi M., Koyama S., Nakazato M., Miyoshi N., Wolf C., Daikuzono N., Tanielian C., Sasaki M., Komiyama M. // J. Clin. Laser. Med. Surg.– 1994.–v.12. – p. 133-138.

Ryazanova O.A., Voloshin I.M., Dubey I.Ya., Dubey L.V., Zozulya V.N. // Annals of the New York Academy of Sciences.– 2008.– v.1130.– p. 293-299.

Ryazanova O.A., Voloshin I.M., Dubey I.Ya., Dubey L.V. and Zozulya V.N. // IFMBE Proceedings.– 2008.- v.20.- p. 397-400.

Wongsinkongman P., Brossi A., Wang H.-K., Bastow K.F., Lee K.-H. // Bioorganic & Medicinal Chemistry.– 2002.– v.10.– p.583-591.

Zozulya V., Blagoi Yu., Lober G., Voloshin I. et al. // Biophys. Chem. –1997-v.65, p. 55-63.

Lakowicz, J.R. 1999. Principles of Fluorescent Spectroscopy 2nd ed. Kluwer Academic/Plenum Press. New York.

Spikes, J.D. & Bommer, J.C. 1991. Chlorophyll and related pigments as photosensitizers in biology and medicine. In Chlorophylls. H. Scheer, Ed.: 1181-1204. CRC Press, Boca Raton, Florida, USA.

Rabinowitch E. // Rewiew of modern physics.–1944.– v. 16(3-4).– p. 226-235.

Röder, B. // J. Photochem. Photobiol, B: Biol.– 1987.– v.1.– p.103-113.

Mansouri S., Fery-Forgues S., Meunier B., Paillous N. // J. Chem. Soc. Perkin Trans. 2 .– 1996.– v.8.– p. 1649-1654.

Mergny J., Phan A., Lacroix L. // FEBS Letters.– 1998.– v.435.- p.74-78.

Löber G., Kittler L. // Studia biophysica.–1978.– v.73(1).– p.25-30.

Seidel C., Schulz A., and Sauer M. // J. Phys. Chem.- 1996.- v.100.– p.5541-555.

Taima H., Yoshioka N., Inoue H. // Org. Biomol. Chem. – 2009.–v. 7.– p. 1176-1183.

Published
2009-06-05
Cited
How to Cite
RyazanovaО. А., VoloshinI. М., Dubey, L. V., & Zozulya, V. N. (2009). Spectroscopic study of pheophorbide-a methyl ether binding to synthetic polynucleotides and DNA. Biophysical Bulletin, 2(23), 20-29. Retrieved from https://periodicals.karazin.ua/biophysvisnyk/article/view/4268