| Autor: |
Backus KM; Department of Chemical Physiology, The Scripps Research Institute. La Jolla, California 92307, USA., Correia BE; Department of Chemical Physiology, The Scripps Research Institute. La Jolla, California 92307, USA., Lum KM; Department of Chemical Physiology, The Scripps Research Institute. La Jolla, California 92307, USA., Forli S; Department of Integrative Structural and Computational Biology, The Scripps Research Institute. La Jolla, California 92307, USA., Horning BD; Department of Chemical Physiology, The Scripps Research Institute. La Jolla, California 92307, USA., González-Páez GE; Department of Molecular and Experimental Medicine, The Scripps Research Institute. La Jolla, California 92307, USA., Chatterjee S; Department of Molecular and Experimental Medicine, The Scripps Research Institute. La Jolla, California 92307, USA., Lanning BR; Department of Chemical Physiology, The Scripps Research Institute. La Jolla, California 92307, USA., Teijaro JR; Department of Immunology and Microbial Science, The Scripps Research Institute. La Jolla, California 92307, USA., Olson AJ; Department of Integrative Structural and Computational Biology, The Scripps Research Institute. La Jolla, California 92307, USA., Wolan DW; Department of Molecular and Experimental Medicine, The Scripps Research Institute. La Jolla, California 92307, USA., Cravatt BF; Department of Chemical Physiology, The Scripps Research Institute. La Jolla, California 92307, USA. |
| Abstrakt: |
Small molecules are powerful tools for investigating protein function and can serve as leads for new therapeutics. Most human proteins, however, lack small-molecule ligands, and entire protein classes are considered 'undruggable'. Fragment-based ligand discovery can identify small-molecule probes for proteins that have proven difficult to target using high-throughput screening of complex compound libraries. Although reversibly binding ligands are commonly pursued, covalent fragments provide an alternative route to small-molecule probes, including those that can access regions of proteins that are difficult to target through binding affinity alone. Here we report a quantitative analysis of cysteine-reactive small-molecule fragments screened against thousands of proteins in human proteomes and cells. Covalent ligands were identified for >700 cysteines found in both druggable proteins and proteins deficient in chemical probes, including transcription factors, adaptor/scaffolding proteins, and uncharacterized proteins. Among the atypical ligand-protein interactions discovered were compounds that react preferentially with pro- (inactive) caspases. We used these ligands to distinguish extrinsic apoptosis pathways in human cell lines versus primary human T cells, showing that the former is largely mediated by caspase-8 while the latter depends on both caspase-8 and -10. Fragment-based covalent ligand discovery provides a greatly expanded portrait of the ligandable proteome and furnishes compounds that can illuminate protein functions in native biological systems. |
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