Apparent non-statistical binding in a ditopic receptor for guanosine

Analysis of stepwise association constants for guests binding to more than one site in a receptor is expected to give a ratio of the first association constant to the second of about 4 : 1 on statistical grounds (since a second guest should have an equal chance of binding to a different site on the same, or a new molecule). Taking account of self-association in our analysis of a system in which the binding sites are close together, we observe a ratio closer to 1 : 1, indicative of non-statistical, or cooperative binding. The longer homologue built around two alkynes displays a very different ratio of stepwise association constants of about 7 : 1. We discuss the origins of this unusual behaviour in terms of steric interactions within the receptors and their corresponding complexes with guanosine derivatives.