Статья

Structural and Functional Analyses of an Allosteric EYA2 Phosphatase Inhibitor That Has On-Target Effects in Human Lung Cancer Cells

Jothi Anantharajan1Experimental Drug Discovery Centre, A*STAR, Singapore, SingaporeHengbo Zhou2Department of Pharmacology, University of Colorado Denver Anschutz Medical Campus, Aurora, ColoradoLingdi Zhang3Department of Biochemistry and Molecular Genetics, University of Colorado Denver Anschutz Medical Campus, Aurora, ColoradoTaylor Hotz2Department of Pharmacology, University of Colorado Denver Anschutz Medical Campus, Aurora, ColoradoMelanie Y. Vincent2Department of Pharmacology, University of Colorado Denver Anschutz Medical Campus, Aurora, ColoradoMelanie A. Blevins3Department of Biochemistry and Molecular Genetics, University of Colorado Denver Anschutz Medical Campus, Aurora, ColoradoAnna Jansson1Experimental Drug Discovery Centre, A*STAR, Singapore, SingaporeJohn Wee Liang Kuan1Experimental Drug Discovery Centre, A*STAR, Singapore, SingaporeElizabeth Yihui Ng1Experimental Drug Discovery Centre, A*STAR, Singapore, SingaporeYee Khoon Yeo1Experimental Drug Discovery Centre, A*STAR, Singapore, SingaporeNithya Baburajendran1Experimental Drug Discovery Centre, A*STAR, Singapore, SingaporeGrace Lin1Experimental Drug Discovery Centre, A*STAR, Singapore, SingaporeAlvin W. Hung1Experimental Drug Discovery Centre, A*STAR, Singapore, SingaporeJoma Joy1Experimental Drug Discovery Centre, A*STAR, Singapore, SingaporeSamarjit Patnaik4National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MarylandJuan Marugán4National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MarylandPratyaydipta Rudra5Department of Biostatistics and Informatics, University of Colorado Anschutz Medical Campus, Aurora, ColoradoDebashis Ghosh5Department of Biostatistics and Informatics, University of Colorado Anschutz Medical Campus, Aurora, ColoradoJeffrey Hill1Experimental Drug Discovery Centre, A*STAR, Singapore, SingaporeThomas H. Keller1Experimental Drug Discovery Centre, A*STAR, Singapore, SingaporeRui Zhao3Department of Biochemistry and Molecular Genetics, University of Colorado Denver Anschutz Medical Campus, Aurora, ColoradoHeide L. Ford2Department of Pharmacology, University of Colorado Denver Anschutz Medical Campus, Aurora, ColoradoCongBao Kang1Experimental Drug Discovery Centre, A*STAR, Singapore, Singapore

2019en

ABI

Аннотация

Abstract EYA proteins (EYA1-4) are critical developmental transcriptional cofactors that contain an EYA domain (ED) harboring Tyr phosphatase activity. EYA proteins are largely downregulated after embryogenesis but are reexpressed in cancers, and their Tyr phosphatase activity plays an important role in the DNA damage response and tumor progression. We previously identified a class of small-molecule allosteric inhibitors that specifically inhibit the Tyr phosphatase activity of EYA2. Herein, we determined the crystal structure of the EYA2 ED in complex with NCGC00249987 (a representative compound in this class), revealing that it binds to an induced pocket distant from the active site. NCGC00249987 binding leads to a conformational change of the active site that is unfavorable for Mg2+ binding, thereby inhibiting EYA2′s Tyr phosphatase activity. We demonstrate, using genetic mutations, that migration, invadopodia formation, and invasion of lung adenocarcinoma cells are dependent on EYA2 Tyr phosphatase activity, whereas growth and survival are not. Further, we demonstrate that NCGC00249987 specifically targets migration, invadopodia formation, and invasion of lung cancer cells, but that it does not inhibit cell growth or survival. The compound has no effect on lung cancer cells carrying an EYA2 F290Y mutant that abolishes compound binding, indicating that NCGC00249987 is on target in lung cancer cells. These data suggest that the NCGC00249987 allosteric inhibitor can be used as a chemical probe to study the function of the EYA2 Tyr phosphatase activity in cells and may have the potential to be developed into an antimetastatic agent for cancers reliant on EYA2′s Tyr phosphatase activity.

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Идентификаторы

DOI: 10.1158/1535-7163.mct-18-1239

Цитирования и источники

Цитирований: 2Использованных источников: 0

Показатели — AkademScholar