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bioRxiv · 10.1101/2022.02.23.481635

Early identification of cooperative fragments for protein-protein interaction stabilization

de Vink, P. J.·Cossar, P. J.·Somsen, B. A.·Ottmann, C. O.·Brunsveld, L.
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Abstract

Modulating protein-protein interactions (PPIs) is an effective approach to drug discovery, with several drugs in the clinic that inhibit PPIs. The orthogonal approach of PPI stabilization has developed slowly, a function of the complicated dynamics of multi-component protein complexes. In contrast to PPI inhibition, where ligand affinity is the driving parameter for efficacy, cooperativity is frequently the directing variable for PPI stabilization. Here we show how STD NMR allows for early-stage detection of cooperativity using the hub protein 14-3-3, a focused library of fragments and several 14-3-3 partner proteins. Further, we validate that the observed enhancement in STD signal is a function of cooperativity of the ternary 14-3-3 complex, using mutagenesis and X-ray crystallography. Additionally, we assess the differential cooperativity of three fragments in a panel of 14-3-3 interaction partners. Finally, we demonstrate how selective 14-3-3 complex formation is a function of cooperativity effects

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de Vink, P. J., Cossar, P. J., Somsen, B. A., Ottmann, C. O., Brunsveld, L.. 2022-02-24. Early identification of cooperative fragments for protein-protein interaction stabilization. https://doi.org/10.1101/2022.02.23.481635

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