Objectives: Design and synthesise AfBP-anchored bisubstrate probes across 3 kinases with cleavable linkers and FRET pairs; establish substrate-displacement readouts that distinguish ATP-site binders from substrate-site competitors; benchmark transferability; implement AfBP-based counterselection; publish kit-style SOPs.
Expected results: A generalisable bisubstrate FRET format with construct-design rules, per-kinase benchmarking datasets and open, transferable SOPs; short methods paper + technical note demonstrating ATP-occlusion vs substrate-displacement discrimination; FAIR-deposited data.
Secondments: UoC: smFRET comparison of apo vs ATP-locked kinases (substrate-cleft geometry) (M10–12; 3 months); CNRS: Assay-based small-molecule hit triage for substrate binders (M28–32; 5 months).
Integration: Complements DC1 by providing the second WP1 “common filter”; supplies WP3 screening cascades and WP4 validation with substrate-site readouts.
Interdisciplinarity: Combines chemical biology, assay engineering, HTS screening and biophysics (smFRET, kinetic profiling).
