title: Allosteric inhibitors of the Eya2 phosphatase are selective and inhibit Eya2-mediated cell migration.
abstract: Eya proteins are essential co-activators of the Six family of transcription factors and contain a unique tyrosine phosphatase domain belonging to the haloacid dehalogenase family of phosphatases. The phosphatase activity of Eya is important for the transcription of a subset of Six1-target genes, and also directs cells to the repair rather than apoptosis pathway upon DNA damage. Furthermore, Eya phosphatase activity has been shown to mediate transformation, invasion, migration, and metastasis of breast cancer cells, making it a potential new drug target for breast cancer. We have previously identified a class of N-arylidenebenzohydrazide compounds that specifically inhibit the Eya2 phosphatase. Herein, we demonstrate that these compounds are reversible inhibitors that selectively inhibit the phosphatase activity of Eya2, but not Eya3. Our mutagenesis results suggest that this class of compounds does not bind to the active site and the binding does not require the coordination with Mg(2+). Moreover, these compounds likely bind within a site on the opposite face of the active site, and function as allosteric inhibitors. We also demonstrate that this class of compounds inhibits Eya2 phosphatase-mediated cell migration, setting the foundation for these molecules to be developed into chemical probes for understanding the specific function of the Eya2 phosphatase and to serve as a prototype for the development of Eya2 phosphatase specific anti-cancer drugs.
authors.affiliation: From the Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, Colorado 80045.
authors.forename: Aaron B
authors.initials: AB
authors.lastname: Krueger
authors.affiliation: the Department of Pharmacology, University of Colorado School of Medicine, Aurora, Colorado 80045.
authors.forename: David J
authors.initials: DJ
authors.lastname: Drasin
authors.affiliation: the National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland 20892, and.
authors.forename: Wendy A
authors.initials: WA
authors.lastname: Lea
authors.affiliation: the Department of Pharmacology, University of Colorado School of Medicine, Aurora, Colorado 80045.
authors.forename: Aaron N
authors.initials: AN
authors.lastname: Patrick
authors.affiliation: the National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland 20892, and.
authors.forename: Samarjit
authors.initials: S
authors.lastname: Patnaik
authors.affiliation: the Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado School of Pharmacy, Aurora, Colorado 80045.
authors.forename: Donald S
authors.initials: DS
authors.lastname: Backos
authors.affiliation: the Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado School of Pharmacy, Aurora, Colorado 80045.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Matheson
authors.affiliation: the National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland 20892, and.
authors.forename: Xin
authors.initials: X
authors.lastname: Hu
authors.affiliation: the National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland 20892, and.
authors.forename: Elena
authors.initials: E
authors.lastname: Barnaeva
authors.affiliation: From the Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, Colorado 80045.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Holliday
authors.affiliation: From the Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, Colorado 80045.
authors.forename: Melanie A
authors.initials: MA
authors.lastname: Blevins
authors.affiliation: the Department of Pharmacology, University of Colorado School of Medicine, Aurora, Colorado 80045.
authors.forename: Tyler P
authors.initials: TP
authors.lastname: Robin
authors.affiliation: From the Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, Colorado 80045.
authors.forename: Elan Z
authors.initials: EZ
authors.lastname: Eisenmesser
authors.affiliation: the National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland 20892, and.
authors.forename: Marc
authors.initials: M
authors.lastname: Ferrer
authors.affiliation: the National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland 20892, and.
authors.forename: Anton
authors.initials: A
authors.lastname: Simeonov
authors.affiliation: the National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland 20892, and.
authors.forename: Noel
authors.initials: N
authors.lastname: Southall
authors.affiliation: the Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado School of Pharmacy, Aurora, Colorado 80045.
authors.forename: Philip
authors.initials: P
authors.lastname: Reigan
authors.affiliation: the National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland 20892, and maruganj@mail.nih.gov.
authors.forename: Juan
authors.initials: J
authors.lastname: Marugan
authors.affiliation: the Department of Pharmacology, University of Colorado School of Medicine, Aurora, Colorado 80045, heide.ford@ucdenver.edu.
authors.forename: Heide L
authors.initials: HL
authors.lastname: Ford
authors.affiliation: From the Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, Colorado 80045, rui.zhao@ucdenver.edu.
authors.forename: Rui
authors.initials: R
authors.lastname: Zhao
mesh_terms: D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D002151:Calorimetry; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology*; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D008274:Magnesium / Q000378:metabolism; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D011485:Protein Binding; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D017386:Sequence Homology, Amino Acid; D013056:Spectrophotometry, Ultraviolet
chemical_list: D004791:Enzyme Inhibitors; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; C106938:EYA2 protein, human; D017027:Protein Tyrosine Phosphatases; D008274:Magnesium
keywords: Anticancer Drug; Enzyme Inhibitor; Migration; Phosphatase; Transcription Coactivator
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