Interaction of Prodan with tubulin. A fluorescence spectroscopic study

The compound 6-propionyl-2-(N,N-dimethyl)-aminonaphthalene (Prodan), an efficient fluorescent probe for proteins, is shown to bind to tubulin. Detailed experiments on fluorescence enhancement, anisotropy and energy transfer were carried out to unravel the nature of Prodan-tubulin interaction and the Prodan-binding site on tubulin. It was found that Prodan binds to tubulin at a rigid site, with a stoichiometry of 1:1 and a dissociation constant of 20 microM. Competition experiments using 1,8-[3H]anilinonaphthalenesulfonate ([3H]ANS) indicated that the three aminonaphthalenes, ANS, bis(1,8-anilinonaphthalenesulfonate) and Prodan probably bind to a common region on the tubulin molecule.