Actinomycin D-Deoxynucleotide Interactions: Binding Isotherms at the Benzenoid and Quinoid Portions of the Drug

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 75; no. 10; pp. 4729 - 4733
• Main Authors: Auer, Henry E., Thompson, Thomas N.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 01-10-1978; National Acad Sciences
• Subjects: Actinomycin; Binding Sites; Biochemistry; Chromophores; Circular Dichroism; Dactinomycin - metabolism; Deoxycytosine Nucleotides - metabolism; Deoxyguanine Nucleotides - metabolism; Deoxyribonucleotides - metabolism; Dichroism; Isotherms; Nucleotides; Temperature; Thermodynamics; Titration; Wavelengths
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.75.10.4729

Titrations of actinomycin D (AMD) with dG and with dG-dC were monitored by circular dichroism at 380 nm and 470 nm. These wavelengths are sensitive predominantly to nucleotide binding processes at the benzenoid and quinoid portions, respectively, of the phenoxazone ring of the drug chromophore [Auer, H. E., Pawlowski-Konopnicki, B. E., Chiao, Y. C. C. & Krugh, T. R. (1978), Biopolymers, 17, 1891-1911.]. The temperature dependence of these isotherms was analyzed by the van't Hoff equation to obtain values for the enthalpy and entropy changes. For dG these are about -11 kcal mol-1 and -20 cal mol-1 deg-1, respectively, for complex formation at both the benzenoid and quinoid sites (1 cal = 4.184 J). The enthalpy and entropy changes for complex formation with dG-dC remain unchanged at the benzenoid site, but both values are more negative at the quinoid site. These results indicate that the additional process of binding C in the intercalated AMD-(dG-dC)2 complex, with respect to the simply stacked AMD-dG2 complex, has distinctive properties at the two sites, reflecting their structural differences. The ability to resolve binding processes at the two sites by circular dichroism has permitted us to suggest assignments for the two 31P magnetic resonance lines from the phosphodiester groups observed in the AMD-(pdG-dC)2 complex [Patel, D. J. (1976) Biochim. Biophys. Acta 442, 98-108].