Change in Allosteric Network Affects Binding Affinities of PDZ Domains: Analysis through Perturbation Response Scanning

Abstract

The allosteric mechanism plays a key role in cellular functions of several PDZ domain proteins (PDZs) and is directly linked to pharmaceutical applications; however, it is a challenge to elaborate the nature and extent of these allosteric interactions. One solution to this problem is to explore the dynamics of PDZs, which may provide insights about how intramolecular communication occurs within a single domain. Here, we develop an advancement of perturbation response scanning (PRS) that couples elastic network models with linear response theory (LRT) to predict key residues in allosteric transitions of the two most studied PDZs (PSD-95 PDZ3 domain and hPTP1E PDZ2 domain). With PRS, we first identify the residues that give the highest mean square fluctuation response upon perturbing the binding sites. Strikingly, we observe that the residues with the highest mean square fluctuation response agree with experimentally determined residues involved in allosteric transitions. Second, we construct the allosteric pathways by linking the residues giving the same directional response upon perturbation of the binding sites. The predicted intramolecular communication pathways reveal that PSD-95 and hPTP1E have different pathways through the dynamic coupling of different residue pairs. Moreover, our analysis provides a molecular understanding of experimentally observed hidden allostery of PSD-95. We show that removing the distal third alpha helix from the binding site alters the allosteric pathway and decreases the binding affinity. Overall, these results indicate that (i) dynamics plays a key role in allosteric regulations of PDZs, (ii) the local changes in the residue interactions can lead to significant changes in the dynamics of allosteric regulations, and (iii) this might be the mechanism that each PDZ uses to tailor their binding specificities regulation.

Author Summary

PDZ domain proteins (PDZs) act as adapters in organizing functional protein complexes. Through dynamic interactions, PDZs play a key role in mediating key cellular functions in the cell, and they are linked to currently challenging diseases including Alzheimer's, Parkinson's and cancer. Moreover, they are associated with allosteric regulations in mediating signaling. Therefore, it is critical to have knowledge of how the allosteric transition occurs in PDZs. We investigate the allosteric response of the two most studied PDZs, PSD-95 and hPTP1E, using the perturbation response scanning (PRS) approach. The method treats the protein as an elastic network and uses linear response theory (LRT) to obtain residue fluctuations upon exerting directed random forces on selected residues. With this efficient and fast approach, we identify the key residues that mediate long-range communication and find the allosteric pathways. Although the structures of PSD-95 and hPTP1E are very similar, our analysis predicts that their allosteric pathways are different. We also observe a significant change in allosteric pathways and a decrease in binding affinity upon removal of the distal α3 helix of PSD-95. This approach enables us to understand how dynamic interactions play an important role in allosteric regulations.