The interactions in solution and in the fibre state of the trypanocidal drug, prothidium dibromide, with DNA have been investigated using a number of biophysical techniques. The binding parameters at 0.11 ionic strength were determined by spectroscopic means. Sedimentation studies show that the drug is able to unwind closed circular DNA in solution, but X-ray diffraction and linear dichroism experiments indicate that it is uncertain whether this unwinding can be attributed to intercalation in the classical sense. On the basis of our results, we propose that the primary mode of binding is "sideways" intercalation, supplemented by electrostatic binding along the sugar-phosphate chains and interstrand binding involving hydrogen bonding.