Extended Data Fig. 1: De novo design and production of Neo-4 molecules.
From: Design of cell-type-specific hyperstable IL-4 mimetics via modular de novo scaffolds
a, Structural alignment of Neo-2 and hIL-4. Residues in hIL-4 at the hIL-4/hIL-4Rα interface that were grafted onto Neo-2 are depicted with sticks, and the corresponding residues on Neo-2 are depicted with lines. b, structural alignment of hNeo-4 and mIL-4. Residues in mIL-4 at the mIL-4/mIL-4Rα interface that were grafted onto hNeo-4 are depicted with sticks, and the corresponding residues on hNeo-4 are depicted with lines. c, Amino acid sequence alignment of Neo-2, designed hNeo-4, and the final evolved hNeo-4 (henceforth denoted hNeo-4), demonstrating the evolution of the hNeo-4 sequence during the development process. Mutations introduced at each round of evolution are marked in red. d, Amino acid sequence alignment of designed hNeo-4, designed mNeo-4 and the final evolved mNeo-4 (henceforth denoted mNeo-4), demonstrating the evolution of the mNeo-4 sequence during the development process. Mutations introduced at each round of evolution are marked in red. mIL-4 cysteine at position 87, depicted in yellow, was not grafted to the hNeo-4 structure. e. SDS-PAGE analysis of purified hNeo-4 and mNeo-4. Experiment was repeated once independently with similar results. f, AlphaFold2 predictions of the structure of hNeo-4 bound to the human type I IL-4 receptor complex (peach) aligned to the IL-4Rα subunit in the experimentally determined crystal structure of the type I hIL-4 complex (gray; PDB ID 3BPL). hNeo-4 is predicted to preserve all three binding interfaces present in the hIL-4 signaling complex.
