Extended Data Fig. 4: GHP-88309-016 target mapping through photoaffinity labeling.
From: Orally efficacious broad-spectrum allosteric inhibitor of paramyxovirus polymerase
GHP-88309-016 target mapping through photoaffinity labeling. a, Structure of GHP-88309-016. b, GHP-88309-016 is bioactive, potently inhibiting MeV and HPIV3 replication without appreciable cytotoxicity (n=3). EC50 values from 4-parameter variable slope regression models are shown with 95% CIs. c, SDS-PAGE fractionation of purified MeV L1708 constructs used in UV crosslinking and BLI studies on 7.5% gels, followed by Coomassie blue staining. Polymerase complexes were purified once in sufficient quantity to meet the needs of the project. d, 2D-schematic of the MeV L protein with locations of known resistance mutations (top) and peptides identified by photoaffinity labeling (bottom; black bars). Cyan, green, yellow, orange, and red depict the RdRP, capping, connector, MTase, and C-terminal domains. e, Homology model of MeV L showing the locations of the peptides crosslinked to GHP-88309-016 (black spheres). Confirmed GHP-88309 resistance sites are shown in red. Peptides 1 and 2 are located on the exterior of the polymerase. f, Only residues of peptide 3 (black) are exposed to the interior channels of the polymerase in proximity of the resistance sites (red). The homology model was based on the coordinates reported for HPIV5 L (PDB: 6v85).
