Proteomic discovery of chemical probes that perturb protein complexes in human cells
Proteomic discovery of chemical probes that perturb protein complexes in human cells
Most human proteins lack chemical probes, and several large-scale and generalizable small-molecule binding assays have been introduced to address this problem. How compounds discovered in such binding-first assays affect protein function, nonetheless, often remains unclear. Here, we describe a function-first proteomic strategy that uses size exclusion chromatography (SEC) to assess the global impact of electrophilic compounds on protein complexes in human cells. Integrating the SEC data with cysteine-directed activity-based protein profiling identifies changes in protein-protein interactions that are caused by site-specific liganding events, including the stereoselective engagement of cysteines in PSME1 and SF3B1 that disrupt the PA28 proteasome regulatory complex and stabilize a dynamic state of the spliceosome, respectively. Our findings thus show how multidimensional proteomic analysis of focused libraries of electrophilic compounds can expedite the discovery of chemical probes with site-specific functional effects on protein complexes in human cells.
Jaeger Martin、DeMeester Kristen、Njomen Evert、Hogg Simon、Rahman Jahan、Whitby Landon、Schafroth Michael、Ogasawara Daisuke、Hanigan Thomas、Simon Gabriel、Abdel-Wahab Omar、Cravatt Benjamin、Saghatelian Alan、Reinhardt Christopher、Won Sang Joon、Lindsey Garrett、Her Hsuan-lin、Germain Jason、Rothamel Katherine、Ghosh Brahma、Lazear Michael、Vartabedian Vincent、Vaughan Joan、Melillo Bruno、Teijaro John、Ahn Kay、Heo Inha、Barbas Sabrina、Dix Melissa、Schreiber Stuart、Li Haoxin、Koo Seong Joo、Yokoyama Minoru、Yeo Gene、Remsberg Jarrett
生物科学研究方法、生物科学研究技术生物化学分子生物学
Jaeger Martin,DeMeester Kristen,Njomen Evert,Hogg Simon,Rahman Jahan,Whitby Landon,Schafroth Michael,Ogasawara Daisuke,Hanigan Thomas,Simon Gabriel,Abdel-Wahab Omar,Cravatt Benjamin,Saghatelian Alan,Reinhardt Christopher,Won Sang Joon,Lindsey Garrett,Her Hsuan-lin,Germain Jason,Rothamel Katherine,Ghosh Brahma,Lazear Michael,Vartabedian Vincent,Vaughan Joan,Melillo Bruno,Teijaro John,Ahn Kay,Heo Inha,Barbas Sabrina,Dix Melissa,Schreiber Stuart,Li Haoxin,Koo Seong Joo,Yokoyama Minoru,Yeo Gene,Remsberg Jarrett.Proteomic discovery of chemical probes that perturb protein complexes in human cells[EB/OL].(2025-03-28)[2025-06-03].https://www.biorxiv.org/content/10.1101/2022.12.12.520090.点此复制
评论