Evidence for partially bound states in cooperative molecular recognition interfaces.

A zinc porphyrin equipped with four amide H-bonding sites provides a rigid molecular receptor for the study of cooperative multipoint binding interactions. The interaction of this receptor with a variety of pyridine ligands bearing zero, one, and two H-bonding sites has been studied using UV/vis absorption, (1)H and (31)P NMR spectroscopy, and isothermal titration calorimetry in five different solvents. The results are analyzed in terms of a bound state that populates an ensemble of different complexes in which zero, one, or two of the potential H-bond interactions are formed. The key parameter that determines the behavior of the system is the product of the association constant for the H-bond interaction, K(H), and the effective molarity for the intramolecular interaction, EM. In the system reported here, EM is 0.1-1 M for all of the intramolecular interactions. For strong H-bonds (large K(H) in nonpolar solvents), all of the interactions are formed in the complex and the fully bound state dominates. In this case, additional binding interactions produce incremental increases in complex stability. However, for weaker H-bonds (small K(H) in polar solvents), the formation of additional interactions does not lead to an increase in the overall stability of the complex, due to the population of partially bound states.