title: ACTH Modulates PTP-PEST Activity and Promotes Its Interaction With Paxillin.
abstract: Adrenocorticotropic hormone (ACTH) treatment has been proven to promote paxillin dephosphorylation and increase soluble protein tyrosine phosphatase (PTP) activity in rat adrenal zona fasciculata (ZF). Also, in-gel PTP assays have shown the activation of a 115-kDa PTP (PTP115) by ACTH. In this context, the current work presents evidence that PTP115 is PTP-PEST, a PTP that recognizes paxillin as substrate. PTP115 was partially purified from rat adrenal ZF and PTP-PEST was detected through Western blot in bioactive samples taken in each purification step. Immunohistochemical and RT-PCR studies revealed PTP-PEST expression in rat ZF and Y1 adrenocortical cells. Moreover, a PTP-PEST siRNA decreased the expression of this phosphatase. PKA phosphorylation of purified PTP115 isolated from non-ACTH-treated rats increased KM and VM . Finally, in-gel PTP assays of immunoprecipitated paxillin from control and ACTH-treated rats suggested a hormone-mediated increase in paxillin-PTP115 interaction, while PTP-PEST and paxillin co-localize in Y1 cells. Taken together, these data demonstrate PTP-PEST expression in adrenal ZF and its regulation by ACTH/PKA and also suggest an ACTH-induced PTP-PEST-paxillin interaction. J. Cell. Biochem. 117: 2170-2181, 2016. © 2016 The Authors. Journal of Cellular Biochemistry Published by Wiley Periodicals, Inc.
authors.affiliation: Department of Biochemistry, School of Medicine, Institute for Biomedical Research (INBIOMED), University of Buenos Aires-CONICET, Buenos Aires, Argentina.
authors.forename: Alejandra Beatriz
authors.initials: AB
authors.lastname: Gorostizaga
authors.affiliation: Department of Biochemistry, School of Medicine, Institute for Biomedical Research (INBIOMED), University of Buenos Aires-CONICET, Buenos Aires, Argentina.
authors.forename: M Mercedes
authors.initials: MM
authors.lastname: Mori Sequeiros Garcia
authors.affiliation: Department of Biochemistry, School of Medicine, Institute for Biomedical Research (INBIOMED), University of Buenos Aires-CONICET, Buenos Aires, Argentina.|Pharmacology Unit, School of Dentistry, University of Buenos Aires, Buenos Aires, Argentina.
authors.forename: Andrea B
authors.initials: AB
authors.lastname: Acquier
authors.affiliation: Institute of Cell Biology and Neurosciences "Prof. E. De Robertis", University of Buenos Aires-CONICET, Buenos Aires, Argentina.
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Lopez-Costa
authors.affiliation: Department of Biochemistry, School of Medicine, Institute for Biomedical Research (INBIOMED), University of Buenos Aires-CONICET, Buenos Aires, Argentina.|Pharmacology Unit, School of Dentistry, University of Buenos Aires, Buenos Aires, Argentina.
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Mendez
authors.affiliation: Department of Biochemistry, School of Medicine, Institute for Biomedical Research (INBIOMED), University of Buenos Aires-CONICET, Buenos Aires, Argentina.
authors.forename: Paula M
authors.initials: PM
authors.lastname: Maloberti
authors.affiliation: Department of Biochemistry, School of Medicine, Institute for Biomedical Research (INBIOMED), University of Buenos Aires-CONICET, Buenos Aires, Argentina.
authors.forename: Cristina
authors.initials: C
authors.lastname: Paz
mesh_terms: D000324:Adrenocorticotropic Hormone / Q000494:pharmacology*; D000818:Animals; D045744:Cell Line, Tumor; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D051379:Mice; D051419:Paxillin / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D054594:Protein Tyrosine Phosphatase, Non-Receptor Type 12 / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D015383:Zona Fasciculata / Q000166:cytology / Q000378:metabolism*
chemical_list: D051419:Paxillin; C496063:Pxn protein, rat; D000324:Adrenocorticotropic Hormone; D017868:Cyclic AMP-Dependent Protein Kinases; D054594:Protein Tyrosine Phosphatase, Non-Receptor Type 12
keywords: ACTH; ADRENAL ZONA FASCICULATA; PAXILLIN; PHOSPHORYLATION; PROTEIN TYROSINE PHOSPHATASE