Fig. 2: Concept, synthesis, and general screening workflow of the pharmacophore-optimized photoaffinity (PhP) library.
From: Mapping protein binding sites by photoreactive fragment pharmacophores
a Primary amines from the Enamine collection were compiled with the SpotXplorer workflow to optimize diversity and coverage against 117 unique 2- and 3-point fragment binding pharmacophores. b HATU coupling in plate-based parallel synthesis and subsequent purification yielded 100 (out of 160) fragments equipped with a diazirine tag. c The screening paradigm employs 10 min of irradiation with the protein target and subsequent MS detection of the anchored protein-fragment complex. d Screening results were categorized by the analysis of their mass spectra as strong binders (>5% labeling), weak binders (1–5% labeling), multiple binders (multiple labeling), oxidized samples (having only protein +16 or multiplied peaks) and non-binders (<1% labeling) samples showing no change in the MS spectra.
