Mapping protein binding sites by photoreactive fragment pharmacophores

• DOI: 10.1038/s42004-024-01252-w
• Publication Date: 2024-07-31T20:04:53Z
• Authors: Péter Ábrányi-Balogh; Dávid Bajusz; Zoltán Orgován; Aaron B. Keeley; László Petri; Nikolett Péczka; Tibor Viktor Szalai; Gyula Pálfy; Márton Gadanecz; Emma K. Grant; Tímea Imre; Tamás Takács; Ivan Ranđelović; Marcell Baranyi; András Marton; Gitta Schlosser; Qirat F. Ashraf; Elvin D. de Araujo; Tamás Karancsi; László Buday; József Tóvári; András Perczel; Jacob T. Bush; György M. Keserű
• Keywords: Diazirine; Fragment (logic); Photoaffinity labeling; Target protein

Fragment screening is a popular strategy of generating viable chemical starting points especially for challenging targets. Although fragments provide better coverage space and they have typically higher chance binding, their weak affinity necessitates highly sensitive biophysical assays. Here, we introduce concept that combines evolutionary optimized fragment pharmacophores with the use photoaffinity handle enables high hit rates by LC-MS-based detection. The sensitivity our protocol was further improved target-conjugated photocatalyst. We designed, synthesized, screened 100 diazirine-tagged against three benchmark therapeutically relevant protein targets different tractability. Our therapeutic included conventional enzyme, first bromodomain BRD4, protein-protein interaction represented oncogenic KRas