Extended Data Fig. 2: 3P10 binds GFRAL at its interface with RET and GFRAL Arg294 is required for 3P10 binding.
From: Antibody-mediated inhibition of GDF15–GFRAL activity reverses cancer cachexia in mice
a, A 2.97 Å crystal structure of 3P10 in complex with GFRAL ECD, with GDF15 docked from 5VZ4.pdb. b, The GFRAL ECD interface with 3P10 shares 5 common residues (Arg294, Thr295, Thr297, Gln298, Ser299) with the GFRAL ECD interface with RET ECD from 6Q2J.pdb, based on structural docking. c, 3P10 HC-Tyr100 and 3P10 LC-Tyr53 in an edge-to-face pi-pi stacking interaction with GFRAL Arg294 (yellow). d, FACS plots of 3P10 binding to cells expressing wildtype GFRAL but not to cells expressing Arg294Glu mutant GFRAL. 25M22-FITC binding to cells expressing both wildtype GFRAL and the Arg294Glu mutant GFRAL are shown as a control for cell surface expression. Data representative of two independent experiments. e, GDF15 reporter gene assay of cells expressing wildtype or Arg294Glu mutant GFRAL incubated with increasing concentrations of 3P10. N = 3 per condition. Data represent mean ± s.d.
