Add time:08/12/2019         Source:sciencedirect.com

Calicheamicin, γ1I, is a remarkable DNA binding-cleaving, enediyne-containing, natural product that exhibits potent antitumor activity. In this study, we used electronic circular dichroism spectroscopy to monitor potential drug-induced DNA conformational changes and DNA induced conformational changes in the calicheamicin aglycone. Three DNA dodecamer sequences were examined: one containing a primary TCCT binding/cleavage site and two dodecamers containing less prominent CTCT and TCTC sites. The binding was monitored by taking advantage of the drug’s unique negative exciton couplet (−313 nm/+275 nm) in phosphate buffer/ethanol 10%. Specifically the CD analysis focused at the longest wavelength region around 313 nm where there is no interference by the positive CD contributions of the DNA. Upon binding at a DNA/drug ratio of 1/1.2 and 1/2.7 a slight red shift from 313 nm to 319 nm was observed. At a ratio of 1/1.2, the CE intensity remained practically unchanged from that of free drug, which indicates no conformational changes in the bound aglycone itself. A larger amount of drug, at a molar ratio of DNA/drug of 1/2.7 but especially at 1/6 and up to 1/10, however, caused a surprisingly distinct decrease in the intensity at this negative CD band and a further small red-shift to 322 nm, evidence for non-specific binding.

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