Novel Allosteric Effectors Targeting Human Transcription Factor TEAD

by Selleckchem | Oct 15 2023

The Hippo pathway is an evolutionary conserved signaling network involved in several cellular regulatory processes. Dephosphorylation and overexpression of Yes-associated proteins (YAPs) in the Hippo-off state are common in several types of solid tumors. YAP overexpression results in its nuclear translocation and interaction with transcriptional enhanced associate domain 1-4 (TEAD1-4) transcription factors. Covalent and non-covalent inhibitors have been developed to target several interaction sites between TEAD and YAP. The most targeted and effective site for these developed inhibitors is the palmitate-binding pocket in the TEAD1-4 proteins. Screening of a DNA-encoded library against the TEAD central pocket was performed experimentally to identify six new allosteric inhibitors. Inspired by the structure of the TED-347 inhibitor, chemical modification was performed on the original inhibitors by replacing secondary methyl amide with a chloromethyl ketone moiety. Various computational tools, including molecular dynamics, free energy perturbation, and Markov state model analysis, were employed to study the effect of ligand binding on the protein conformational space. Four of the six modified ligands were associated with enhanced allosteric communication between the TEAD4 and YAP1 domains indicated by the relative free energy perturbation to original molecules. Phe229, Thr332, Ile374, and Ile395 residues were revealed to be essential for the effective binding of the inhibitors.

Comments:

The Hippo pathway is a signaling network that is highly conserved across evolution and plays a crucial role in regulating various cellular processes. In several types of solid tumors, there is a common occurrence of dephosphorylation and overexpression of Yes-associated proteins (YAPs) in the Hippo-off state. This leads to the nuclear translocation of YAP and its interaction with transcriptional enhanced associate domain 1-4 (TEAD1-4) transcription factors.

Researchers have developed inhibitors, both covalent and non-covalent, to target the interaction between TEAD and YAP. Among these inhibitors, the most effective and targeted site is the palmitate-binding pocket in the TEAD1-4 proteins. To identify new allosteric inhibitors, a screening of a DNA-encoded library against the TEAD central pocket was conducted experimentally, resulting in the discovery of six new inhibitors.

Building upon the structure of the TED-347 inhibitor, chemical modifications were made to the original inhibitors by replacing the secondary methyl amide with a chloromethyl ketone moiety. To study the impact of ligand binding on protein conformational space, various computational tools such as molecular dynamics, free energy perturbation, and Markov state model analysis were employed.

The computational analysis revealed that four of the modified ligands exhibited enhanced allosteric communication between the TEAD4 and YAP1 domains, as indicated by their relative free energy perturbation compared to the original molecules. The residues Phe229, Thr332, Ile374, and Ile395 were identified as essential for the effective binding of these inhibitors.

Overall, this research provides insights into the development and optimization of allosteric inhibitors targeting the TEAD1-4 proteins, which could have potential applications in regulating the Hippo pathway and combating certain types of solid tumors.