Binding of Hoechst with nucleic acids using fluorescence spectroscopy英文文献资料.docVIP

Vol.2, No.4, 443-447 (2011) Journal of Biophysical Chemistry doi:10.4236/jbpc.2011.24052 Binding of Hoechst with nucleic acids using fluorescence spectroscopy Nikolai Vekshin Institute of Cell Biophysics of Russian Academy of Sciences, Pushchino, Moscow Region, Russia. nvekshin@rambler.ru Received 2 August 2011; revised 14 September 2011; accepted 27 September 2011. ABSTRACT Full understanding of the molecular mechanisms of action of heterocyclic antibiotics at low concentrations on DNA and RNA can optimize the use of these com- pounds and their analogues as medical drugs. It has been shown that polarity of environment around Hoechst 33342 is almost unchanged while sorption of this fluorescent dye on a sur- face of the hairpin oligonucleotide HP1, t-RNA and DNA. At small concentrations, this dye, ad- sorbed on the surface of DNA, RNA or HP1, does not show any specificity to certain nucleotides. In the case of unwound sites of DNA or HP1, it can bind inside, but without the intercalation stacking with nucleotides. The energy transfer from nucleotide chromophores to Hoechst is absent due to their remoteness and also “bad” (non-stacking) orientation. The mutual fluores- cence quenching of Hoechst by actinomycin D (AMD) and, vice versa, of 7-amino-actinomycin D (7AAMD) by Hoechst in DNA and HP1 is ob- served. It is due to dynamic deactivation and mutual replacing in binding sites. Hoechst dyes 33342 and 33258 are closely related bis- benzimides. Hoechst 33258 is 2’-(4-hydroxyphenyl)-5- [5-(4-methylpiperazine-1-yl) benzimidazo-2-yl]-benzimi- dazole. Hoechst 33342 contains additional ethyl group. This group renders Hoechst 33342 a bit more hydropho- bic than Hoechst 33258. Both dyes have the absorption band at 350 nm and the emission band in the “blue” re- gion. They are non-intercalating fluorescent probes for non-covalent labeling of DNA. They are used in fluores