Fragment-based exploration of the 14-3-3/Amot-p130 interface

Federica Centorrino; Blaž Andlovic; Peter Cossar; Luc Brunsveld; Christian Ottmann

doi:10.1016/j.crstbi.2021.12.003

Fragment-based exploration of the 14-3-3/Amot-p130 interface

Curr Res Struct Biol. 2021 Dec 29:4:21-28. doi: 10.1016/j.crstbi.2021.12.003. eCollection 2022.

Authors

Federica Centorrino¹, Blaž Andlovic¹, Peter Cossar¹, Luc Brunsveld¹, Christian Ottmann¹

Affiliation

¹ Laboratory of Chemical Biology, Department of Biomedical Engineering and Institute for Complex Molecular Systems, Eindhoven University of Technology, P. O. Box 513, 5600 MB, Eindhoven, the Netherlands.

Abstract

The modulation of protein-protein interactions (PPIs) has developed into a well-established field of drug discovery. Despite the advances achieved in the field, many PPIs are still deemed as 'undruggable' targets and the design of PPIs stabilizers remains a significant challenge. The application of fragment-based methods for the identification of drug leads and to evaluate the 'tractability' of the desired protein target has seen a remarkable development in recent years. In this study, we explore the molecular characteristics of the 14-3-3/Amot-p130 PPI and the conceptual possibility of targeting this interface using X-ray crystallography fragment-based screening. We report the first structural elucidation of the 14-3-3 binding motif of Amot-p130 and the characterization of the binding mode and affinities involved. We made use of fragments to probe the 'ligandability' of the 14-3-3/Amot-p130 composite binding pocket. Here we disclose initial hits with promising stabilizing activity and an early-stage selectivity toward the Amot-p130 motifs over other representatives 14-3-3 partners. Our findings highlight the potential of using fragments to characterize and explore proteins' surfaces and might provide a starting point toward the development of small molecules capable of acting as molecular glues.

Keywords: 14-3-3 /protein-protein interactions stabilizers; AIP4, Atrophin-1 interacting protein 4; Amot, Angiomotin; Amot-p130; AmotL1/2, Angiomotin-like 1/2; FBDD, Fragment-based drug discovery; FP, Fluorescence polarization; Fragment-based drug discovery; Lats 1/2, Large tumor suppressor 1/2; Ligandability; MST, Microscale thermophoresis; PPI, Protein-protein interaction; PTMs, post-translational modifications; X-ray crystallography; YAP1, Yes-associated protein 1.