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