Fig 1.
Screening of fragment library.
(a) Summary of results from screening of Maybridge fragment library, using thermal stability shift assay. (b) A representative normalized graph showing a shift in thermal stability of the protein TEAD, when a fragment from the library binds to and stabilizes it.
Fig 2.
Structure of TEAD-fragment complex.
(a) Overall structure of two TEAD molecules (purple) in complex with hit fragment 1 (green). (b) Overlay of the TEAD molecule from our structure (purple) with that of the human TEAD2 structure (pink, PDB code: 3L15)[39], shows no conformational changes induced by the binding of fragment 1. (c) Overlay of our structure with the mouse TEAD-YAP complex structure (PDB code: 3JUA)[37], reveals that the benzene rings of mYAP Phe54 (yellow sticks) and fragment 1 (green sticks) occupy the same hydrophobic groove on the TEAD surface (grey). (d) Fragment 1 (green sticks) binds to two molecules of TEAD and mainly form hydrophobic interactions with the surface residues of TEAD (grey and beige sticks). The carboxyl group of the fragment also forms hydrogen bonds (black dash) with the side chain of Lys369.
Table 1.
Data collection and refinement statistics for TEAD-fragment complex.
Fig 3.
Binding affinity and efficacy of fragment 1.
(a) Calorimetric titration of fragment 1 with mTEAD4 protein. The data is fitted to a standard single-binding site model. (b) Cell-based luciferase assay showing a decrease in firefly luciferase signal with increasing concentrations of fragment 1. The data suggests that fragment 1 interferes with the transcriptional activity of TEAD, probably by disrupting the interactions between YAP/TAZ and TEAD.
Fig 4.
Structure-activity relationship study of fragment derivatives.
(a) Chemical structures of the hit fragment 1 and four rationally designed fragment derivatives (2 to 5). (b) Calorimetric titration of compound 5 with mTEAD4 protein. The data shows weak binding of the compound to the protein.
Fig 5.
Computational ligand-mapping identifies cryptic binding sites in TEAD.
(a) The computationally identified high-density benzene sites (mesh) in human TEAD1 (grey surface; PDB code: 3KYS) correspond to the interaction site of Fragment 1 (green sticks) and also to sites of hydrophobic residues in YAP (yellow cartoon; hydrophobic residues shown as sticks). Additional sites around Fragment 1 are identified and could serve as anchors for optimizing Fragment 1. (b) Computational docking of compound 6 (inset; green sticks) to human TEAD1. (c) Calorimetric titration of compound 6 with mTEAD4 protein. The data shows sub-millimolar range binding affinity, comparable to Fragment 1.