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The degree of binding of "33258 Hoechst" to DNA and nucleohistone has been determined by equilibrium dialysis and the properties of the complexes have been followed by different optical and electro-optical methods, after determining the orientation of the main transition moments within the dye molecule. The binding isotherm was found composed of a Langmuir-type and of a strongly cooperative component. The existence of two bound species yielded a continuous variation of most of the properties of the complexes studied as the amount of binding increased, while the hydrodynamic properties of the macromolecules were not affected. At low binding, the strongly bound dye molecules appeared to bind to highly fluorescent sites with their long axis oriented at 45 degree to the helix axis. As the binding proceeds, less fluorescent sites are cooperatively occupied and the inclination of these ligand molecules becomes closer to that of the base planes. These results are compatible with the formation of two external complexes with the double helical structure.

Physico-chemical study of the complexes of ‘33258 Hoechst’ with DNA and nucleohistone