An Exploration of Chemical Properties Required for Cooperative Stabilization of the 14-3-3 Interaction with NF-κB-Utilizing a Reversible Covalent Tethering Approach

Protein-protein modulation has emerged as a proven approach to drug discovery. While significant progress has been gained in developing protein- protein interaction (PPI) inhibitors, the orthogonal approach of PPI stabilization lacks established methodologies for drug design. Here, we report the systematic bottom-up development of a reversible covalent PPI stabilizer. An imine bond was employed to anchor the stabilizer at the interface of the 14-3-3/p65 complex, leading to a molecular glue that elicited an 81-fold increase in complex stabilization. Utilizing protein crystallography and biophysical assays, we deconvoluted how chemical properties of a stabilizer translate to structural changes in the ternary 14-3-3/p65/molecular glue complex. Furthermore, we explore how this leads to high cooperativity and increased stability of the complex.

Automated summary

This study describes the systematic bottom-up development of a reversible covalent PPI stabilizer and investigates how its chemical properties affect the structural changes in the ternary 14-3-3/p65/molecular glue complex, ultimately leading to increased stability and high cooperativity.