A Chemoproteomic Strategy for Direct and Proteome-Wide Covalent Inhibitor Target-Site Identification

• DOI: 10.1021/jacs.8b07911
• Publication Date: 2018-12-06T05:29:21Z
• Authors: Christopher M. Br...; Baishan Jiang; Scott B. Ficarro; Zainab M. Doctor; Jared L. Johnson; Joseph D. Card; Sindhu Carmen Siv...; William M. Alexander; Tomer M. Yaron; Charles J. Murphy; Nicholas P. Kwiat...; Tinghu Zhang; Lewis C. Cantley; Nathanael S. Gray; Jarrod A. Marto
• Keywords: Proteome; Drug target; Drug Development

Despite recent clinical successes for irreversible drugs, potential toxicities mediated by unpredictable modification of off-target cysteines represents a major hurdle expansion covalent drug programs. Understanding the proteome-wide binding profile inhibitors can significantly accelerate their development; however, current mass spectrometry strategies typically do not provide direct, amino acid level readout activity complex, selective inhibitors. Here we report development CITe-Id, novel chemoproteomic approach that employs pharmacologic as enrichment reagents in combination with an optimized proteomic platform to directly quantify dose-dependent at cysteine-thiols across proteome. CITe-Id analysis our CDK inhibitor THZ1 identified several unexpected kinases, including previously unannotated cysteine (C840) on understudied kinase PKN3. These data streamlined JZ128 new Using probe compound, PKN3 substrates, thus offering initial molecular view cellular activity. provides powerful complement platforms characterize selectivity inhibitors, identify new, pharmacologically addressable cysteine-thiols, and inform structure-based design