An Exploration
of Chemical Properties Required for
Cooperative Stabilization of the 14-3‑3 Interaction with NF-κBUtilizing
a Reversible Covalent Tethering Approach
posted on 2021-06-02, 15:09authored byMadita Wolter, Dario Valenti, Peter J. Cossar, Stanimira Hristeva, Laura M. Levy, Thorsten Genski, Torsten Hoffmann, Luc Brunsveld, Dimitrios Tzalis, Christian Ottmann
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.