title: Cell-active dual specificity phosphatase inhibitors identified by high-content screening.
abstract: Phosphorylation of extracellular signal-regulated kinase (Erk) is tightly controlled by dual specificity phosphatases (DSPases), but few inhibitors of Erk dephosphorylation have been identified. Using a high-content, fluorescence-based cellular assay and the National Cancer Institute's 1990 agent Diversity Set, we identified ten compounds (0.5%) that significantly increased phospho-Erk cytonuclear differences in intact cells. Three of the ten positive compounds inhibited the mitogen-activated protein kinase phosphatase-3 (MKP-3/PYST-1) in vitro without affecting VHR or PTP1B phosphatases. The most potent inhibitor of MKP-3 had an IC50 of < 10 microM and inhibited MKP-3 in a novel, fluorescence-based multiparameter chemical complementation assay. These results suggest that the phospho-Erk nuclear accumulation assay may be a useful tool to discover DSPase inhibitors with biological activity.
authors.affiliation: Department of Pharmacology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.
authors.forename: Andreas
authors.initials: A
authors.lastname: Vogt
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Cooley
authors.forename: Marni
authors.initials: M
authors.lastname: Brisson
authors.forename: Michael G
authors.initials: MG
authors.lastname: Tarpley
authors.forename: Peter
authors.initials: P
authors.lastname: Wipf
authors.forename: John S
authors.initials: JS
authors.lastname: Lazo
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001572:Benzofurans / Q000494:pharmacology; D054642:Dual Specificity Phosphatase 6 / Q000037:antagonists & inhibitors; D054637:Dual-Specificity Phosphatases / Q000037:antagonists & inhibitors*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D006367:HeLa Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D051379:Mice; D041681:NIH 3T3 Cells; D020687:cdc25 Phosphatases / Q000037:antagonists & inhibitors
chemical_list: D000970:Antineoplastic Agents; D001572:Benzofurans; D007093:Imidazoles; C483753:NSC 357756; D048049:Extracellular Signal-Regulated MAP Kinases; D054642:Dual Specificity Phosphatase 6; D054637:Dual-Specificity Phosphatases; D020687:cdc25 Phosphatases