title: Identification of a selective small-molecule inhibitor series targeting the eyes absent 2 (Eya2) phosphatase activity.
abstract: Eya proteins are essential coactivators of the Six family of homeobox transcription factors and also contain a unique protein tyrosine phosphatase activity, belonging to the haloacid dehalogenase family of phosphatases. The phosphatase activity of Eya is important for a subset of Six1-mediated transcription, making this a unique type of transcriptional control. It is also responsible for directing cells to the repair instead of apoptosis pathway upon DNA damage. Furthermore, the phosphatase activity of Eya is critical for transformation, migration, invasion, and metastasis of breast cancer cells. Thus, inhibitors of the Eya phosphatase activity may be antitumorigenic and antimetastatic, as well as sensitize cancer cells to DNA damage-inducing therapies. In this article, we identified a previously unknown chemical series using high-throughput screening that inhibits the Eya2 phosphatase activity with IC(50)s ranging from 1.8 to 79 µM. Compound activity was confirmed using an alternative malachite green assay and H2AX, a known Eya substrate. Importantly, these Eya2 phosphatase inhibitors show specificity and do not significantly inhibit several other cellular phosphatases. Our studies identify the first selective Eya2 phosphatase inhibitors that can potentially be developed into chemical probes for functional studies of Eya phosphatase or into anticancer drugs in the future.
authors: Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, CO 80045.
authors: Aaron B
authors: AB
authors: Krueger
authors: NIH Chemical Genomics Center, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD 20892.
authors: Seameen J
authors: SJ
authors: Dehdashti
authors: NIH Chemical Genomics Center, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD 20892.
authors: Noel
authors: N
authors: Southall
authors: NIH Chemical Genomics Center, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD 20892.
authors: Juan J
authors: JJ
authors: Marugan
authors: NIH Chemical Genomics Center, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD 20892.
authors: Marc
authors: M
authors: Ferrer
authors: Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, CO 80045.
authors: Xueni
authors: X
authors: Li
authors: Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, CO 80045.|Department of Pharmacology, University of Colorado School of Medicine, Aurora, CO 80045.|Department of Ob/Gyn, University of Colorado School of Medicine, Aurora, CO 80045.
authors: Heide L
authors: HL
authors: Ford
authors: NIH Chemical Genomics Center, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD 20892.
authors: Wei
authors: W
authors: Zheng
authors: Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, CO 80045.
authors: Rui
authors: R
authors: Zhao
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000737:chemistry*; D005452:Fluoresceins / Q000737:chemistry; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D007700:Kinetics; D008904:Miniaturization; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D010659:Phenylhydrazines / Q000737:chemistry; D010749:Phosphoprotein Phosphatases / Q000737:chemistry; D000071636:Protein Phosphatase 2C; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1 / Q000737:chemistry; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors* / Q000737:chemistry; D054852:Small Molecule Libraries; D013050:Spectrometry, Fluorescence
chemical_list: C032079:3-O-methylfluorescein phosphate; D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D005452:Fluoresceins; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D010659:Phenylhydrazines; D054852:Small Molecule Libraries; D010749:Phosphoprotein Phosphatases; D000071636:Protein Phosphatase 2C; C106938:EYA2 protein, human; C516607:PTPN1 protein, human; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D017027:Protein Tyrosine Phosphatases