PHOSPHORESCENCE AND OPTICALLY DETECTED MAGNETIC-RESONANCE STUDIES OF ECHINOMYCIN DNA COMPLEXES

Echinomycin complexes with polymeric DNAs and model duplex oligonucleotides have been studied by low-temperature phosphorescence and optical detection of triplet-state magnetic resonance (ODMR) spectroscopy, with the quinoxaline chromophores of the drug used as intrinsic probes. Although not optically resolved, plots of ODMR transition frequencies versus monitored wavelength revealed heterogeneity in the phosphorescence emission of echinomycin, which was ascribed to the presence of two distinct quinoxaline triplet-state environments (referred to as the blue and red triplet states of echinomycin in this report). We think that a likely origin of the two triplet states of echinomycin is the occurrence of two or more distinct conformations of the drug in aqueous solutions. Spectroscopically observed perturbations of the triplet-state properties of echinomycin such as the phosphorescence emission spectrum, phosphorescence lifetime, ODMR spectrum, and zero-field splitting (zfs) energies were investigated upon drug binding to the double-stranded alternating copolymers poly(dG-dC)·poly(dG-dC) [abbreviated as poly[d(G-C)2]] and poly(dA-dT)·poly(dA-dT) [abbreviated as poly[d(A-T)2]], the homopolymer duplexes poly(dG)·poly(dC) [abbreviated as poly(dG·dC)] and poly(dA)·poly(dT) [abbreviated as poly(dA·dT)], and the natural DNAs from Escherichia coli, Micrococcus lysodeikticus, and calf thymus. Echinomycin bisintercalation complexes with the self-complementary oligonucleotides d(ACGT), d(CGTACG), and d(ACGTACGT), which are thought to model drug binding sites, were also investigated. Phosphorescence and ODMR spectroscopic results indicate that the quinoxaline chromophores of the drug are involved in aromatic stacking interactions in complexes with the natural DNAs as evidenced by red shifts in the phosphorescence 0,0 band of the drug, a small but significant reduction in the phosphorescence lifetime of the red triplet state, and reduction of the zfs D-value of both the blue and red triplet states upon drug complexation. These changes in the triplet-state properties of echinomycin are consistent with stacking interactions that increase the polarizability of the quinoxaline environment. The extent of the reduction of the D parameter for the red triplet state upon complexation with the polymeric DNAs was found to correlate with the binding affinities measured for these targets [Wakelin, L. P. G., & Waring, M. J. (1976) Biochem. J. 157, 721-740], with the single exception of the drug-poly [d(G-C)2] complex, for which an increase in the D-value was noted. In addition, upon drug binding to the natural DNAs, there is a reversal of signal polarity in the ODMR spectra of the red triplet state. Among the synthetic DNA polymers investigated, a reversal of ODMR signal polarity was found only with the echinomycin-poly[d(A-T)2] complex. The polarity reversals apparently result from changes in the triplet sublevel decay constants upon binding to DNA. Echinomycin binding to the duplex oligonucleotides employed in this work gave results similar to those obtained with the natural DNAs. In relation to the other DNA complexes, the echinomycin-d(CGTACG) complex exhibited significantly enhanced ODMR signals for a blue-shifted triplet state, which were ascribed to solvent-exposed quinoxaline rings at either end of the oligonucleotide lattice as deduced from X-ray diffraction studies [Ughetto, G., et al. (1985) Nucleic Acids Res. 13, 2305-2323]. © 1990, American Chemical Society. All rights reserved.