FLUORESCENCE POLARIZATION STUDIES OF THE BINDING OF BMS-181176 TO DNA

The DNA binding of EMS 181176, an antitumor antibiotic derivative of rebeccamycin was characterized by DNA unwinding assays, as well as by fluorescence emission and polarization spectroscopic techniques. Unwinding and rewinding of supercoiled DNA was interpreted in terms of intercalation of EMS 181176 into DNA. EMS 181176 shows an enhanced fluorescence emission upon binding to the AT sequence and no enhancement upon binding to the GC sequence. EMS 181176 appears to be a weaker binder to poly(dAdT).poly(dAdT) compared to doxorubicin and ethidium bromide. When bound to DNA the rotational motion of EMS 181176 is substantially decreased as evident from the increase in fluorescence polarization. EMS 181176 exhibits a range of binding strengths depending on the DNA This is demonstrated by the Acridine Orange displacement assay using fluorescence polarization