Wang, YujiaDix, Melissa M.Bianco, GiuliaRemsberg, Jarrett R.Lee, Hsin-YuKalocsay, MarianGygi, Steven P.Forli, StefanoVite, GregoryLawrence, R. MichaelParker, Christopher G.Cravatt, Benjamin F.2022-08-182019-10-28Wang, Yujia, Melissa M. Dix, Giulia Bianco, Jarrett R. Remsberg, Hsin-Yu Lee, Marian Kalocsay, Steven P. Gygi, Stefano Forli, Gregory Vite, R Michael Lawrence, Christopher G. Parker, and Benjamin F .Cravatt. 2019. Expedited Mapping of the Ligandable Proteome Using Fully Functionalized Enantiomeric Probe Pairs. Nature Chemistry 11, no. 12: 1113-123.1755-43301755-4349https://nrs.harvard.edu/URN-3:HUL.INSTREPOS:37373016A fundamental challenge in chemical biology and medicine is to understand and expand the fraction of the human proteome that can be targeted by small molecules. We recently described a strategy that integrates fragment-based ligand discovery with chemical proteomics to furnish global portraits of reversible small molecule-protein interactions in human cells. Excavating clear structure-activity relationships from these “ligandability” maps, however, was confounded by the distinct physicochemical properties and corresponding overall protein-binding potential of individual fragments. Here, we describe a compelling solution to this problem by introducing a next-generation set of fully functionalized fragments (FFFs) differing only in absolute stereochemistry. Using these enantiomeric probe pairs, or “enantioprobes”, we identify numerous stereoselective protein-fragment interactions in cells and show that these interactions occur at functional sites on proteins from diverse classes. Our findings thus indicate that incorporating chirality into FFF libraries provides a robust and streamlined method to discover ligandable proteins in cells.en-USGeneral ChemistryGeneral Chemical EngineeringJournal Article2022-08-1810.1038/s41557-019-0351-5