The Role of Flexibility and Conformational Selection in the Binding Promiscuity of PDZ Domains

In molecular recognition, it is often the case that ligand binding is coupled to conformational change in one or both of the binding partners. Two hypotheses describe the limiting cases involved; the first is the induced fit and the second is the conformational selection model. The conformational selection model requires that the protein adopts conformations that are similar to the ligand-bound conformation in the absence of ligand, whilst the induced-fit model predicts that the ligand-bound conformation of the protein is only accessible when the ligand is actually bound. The flexibility of the apo protein clearly plays a major role in these interpretations. For many proteins involved in signaling pathways there is the added complication that they are often promiscuous in that they are capable of binding to different ligand partners. The relationship between protein flexibility and promiscuity is an area of active research and is perhaps best exemplified by the PDZ domain family of proteins. In this study we use molecular dynamics simulations to examine the relationship between flexibility and promiscuity in five PDZ domains: the human Dvl2 (Dishevelled-2) PDZ domain, the human Erbin PDZ domain, the PDZ1 domain of InaD (inactivation no after-potential D protein) from fruit fly, the PDZ7 domain of GRIP1 (glutamate receptor interacting protein 1) from rat and the PDZ2 domain of PTP-BL (protein tyrosine phosphatase) from mouse. We show that despite their high structural similarity, the PDZ binding sites have significantly different dynamics. Importantly, the degree of binding pocket flexibility was found to be closely related to the various characteristics of peptide binding specificity and promiscuity of the five PDZ domains. Our findings suggest that the intrinsic motions of the apo structures play a key role in distinguishing functional properties of different PDZ domains and allow us to make predictions that can be experimentally tested. Author Summary: Proteins that are capable of binding to many different ligands are said to have broad specificity. This is sometimes also referred to as promiscuity. Whether a protein is promiscuous or not can sometimes be readily explained by the structure of the protein and the ligand in terms of electrostatic and steric effects. Sometimes however, this simple interpretation can struggle to explain the experimentally observed data. A prominent case in point is the PDZ domains. These small protein domains bind to unstructured regions of other proteins and are involved in many signaling pathways. Some PDZ domains appear to be more promiscuous than others, but this has been difficult to explain purely on the basis of the composition of residues in the binding groove. In this work we examine the dynamics and conformational flexibility of five key PDZ domains and demonstrate that despite similar folds, these proteins can exhibit quite different dynamics. Furthermore the difference in the dynamic behavior appears to correlate with the observed promiscuity. Our findings suggest that knowledge of the dynamic behavior of the PDZs can be used to rationalize the extent of expected promiscuity. Such knowledge will be critical for drug design against PDZ domains.