Discovery and pharmacophoric characterization of chemokine network inhibitors using phage-display, saturation mutagenesis and computational modelling
Vales S., Kryukova J., Chandra S., Smagurauskaite G., Payne M., Clark CJ., Hafner K., Mburu P., Denisov S., Davies G., Outeiral C., Deane CM., Morris GM., Bhattacharya S.
CC and CXC-chemokines are the primary drivers of chemotaxis in inflamma- tion, but chemokine network redundancy thwarts pharmacological interven- tion. Tick evasins promiscuously bind CC and CXC-chemokines, overcoming redundancy. Here we show that short peptides that promiscuously bind both chemokine classes can be identified from evasins by phage-display screening performed with multiple chemokines in parallel. We identify two conserved motifs within these peptides and show using saturation-mutagenesis phage- display and chemotaxis studies of an exemplar peptide that an anionic patch in the first motif and hydrophobic, aromatic and cysteine residues in the second are functionally necessary. AlphaFold2-Multimer modelling suggests that the peptide occludes distinct receptor-binding regions in CC and in CXC-chemo- kines, with the first and second motifs contributing ionic and hydrophobic interactions respectively. Our results indicate that peptides with broad- spectrum anti-chemokine activity and therapeutic potential may be identified from evasins, and the pharmacophore characterised by phage display, saturation mutagenesis and computational modelling.