Proteomic discovery of chemical probes that perturb protein complexes in human cells
Ontology highlight
ABSTRACT: Most proteins in the human proteome 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.
INSTRUMENT(S): Orbitrap Fusion
ORGANISM(S): Homo Sapiens (human)
TISSUE(S): B Cell, Cell Suspension Culture, Epithelial Cell, Cell Culture
DISEASE(S): Prostate Adenocarcinoma,Colon Cancer
SUBMITTER: Jarrett Remsberg
LAB HEAD: Benjamin F. Cravatt
PROVIDER: PXD029655 | Pride | 2023-04-20
REPOSITORIES: Pride
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