Interaction of Antioxidant Flavonoids with tRNA: Intercalation or External Binding and Comparison with Flavonoid-DNA Adducts

Antioxidants are essential to good health. Flavonoids are powerful antioxidants, and prevent DNA damage. The antioxidative protections are related to their binding modes to a DNA duplex and complexation with free radicals in vivo. Recently we reported the interaction of flavonoids with DNA in vitro (Kanakis et al., J. Biomol. Struct. Dyn. 22, 719–724, 2005), where polyphenol different binding modes were discussed. The aim of this study was to examine the interaction of transfer RNA with quercetin (que), kaempferol (kae), and delphinidin(del) in aqueous solution at physiological conditions and to make a comparison with the corresponding pigment–DNA adducts. Constant tRNA concentration (6.25 mM) and various drug/RNA(phosphate) molar ratios of 1/48 to 1/8 were used. FTIR and UV-visible difference spectroscopic methods have been applied to determine the drug binding mode, the binding constants, and the effects of drug complexation on the stability and conformation of tRNA duplex. Both intercalative and external binding modes were observed. Structural analysis showed que, kae, and a del intercalate tRNA duplex with minor external binding to the major or minor groove and the backbone phosphate group with overall binding constants K _que = 4.80 × 10⁴ M⁻¹,K _kae = 4.65 × 10⁴ M⁻¹, and K _del = 9.47 × 10⁴ M⁻¹. The stability of adduct formation is in the order of del > que > kae. A comparison with flavonoids–DNA adducts showed both intercalation and external bindings with the stability order K _que = 7.25 × 10⁴ M⁻¹, K _kae = 3.60 × 10⁴ M⁻¹, and K _del = 1.66 × 10⁴ M⁻¹. Low flavonoid concentration induces helical stabilization, whereas high pigment content causes helix opening. A partial Bto A–DNA transition occurs at high drug concentration, while tRNA remains in the A-family structure.