Objectives: Reconstitute EGFR and LTK (SMALPs) and soluble BRAF complexes; capture AfBP-locked conformations; map dimer and substrate interfaces; identify hotspot residues that report on interface engagement; cross-validate structural classes with smFRET states and feed geometric constraints back to WP1 assay design.
References: ³² Barniol-Xicota, M. et al. Commun. Biol. 4, 218 (2021); ³³ Barniol-Xicota, M. et al. J. Am. Chem. Soc. 140, 14557–14561 (2018); ³⁴ Merz, M. L. et al. Nat. Chem. Biol. 20, 624–633 (2024); ³⁵ Kong, X.-D. et al. Nat. Biomed. Eng. 4, 560–571 (2020); ³⁶ Kale, S. S. et al. Nat. Chem. 10, 715–723 (2018); ³⁷ Bosc, N. et al. ACS Chem. Biol. 15, 1566–1574 (2020).
Expected results: Medium-resolution cryo-EM maps and focused classifications of low-occupancy interface states; a hotspot/interfacial-geometry map for all three kinases; a methodological paper on SMALP-enabled cryo-EM for dynamic kinase assemblies plus FAIR-deposited structural classes and metadata.
Secondments: UPF: SMALP membrane-reconstitution workflows (M19–22; 4 months); INS: In-cellulo validation (M30–32; 3 months)
Integration: Supplies structural determinants guiding WP1 assay geometry, calibrates WP2 smFRET interpretations and defines actionable interface hotspots for WP3 ligand optimisation.
Interdisciplinarity: Combines cryo-EM, membrane-protein biophysics (SMALPs) and integration with smFRET and cellular validation.
