S-(N-dansylaminoethyl)-6-mercaptoguanosine as a fluorescent probe for the uridine transport system in human erythrocytes.

A fluorescent derivative of 6-mercaptoguanosine, S-(N-dansylaminoethyl)-6-mercaptoguanosine, was synthesized, and found to be a strong inhibitor of the uridine transport system of erythrocyte (Ki approximately 0.3 microM). The emission spectrum of this compound has peaks at 400 and 550 nm. The emission at 550, but not that a 400 nm, in environment-sensitive. A method was devised for preparing a suspension of erythrocyte-membrane fragments with sufficiently low light scattering so that a detailed study could be made of the fluorescence of the probe when bound to membranes. Direct binding measurements showed the existence of a tight binding site, with a dissociation constant of the same order of magnitude as the inhibition constant. Binding of probe and substrate are not mutually exclusive, but the fluorescence and affinity of the bound probe are sensitive to the presence of uridine. The emission spectrum suggests that the bound probe penetrates into the bilayer region of the membrane.