Inhibition of serine-threonine protein phosphatases decreases barrier function of rat pulmonary microvascular endothelial cells

Journal of Cellular Physiology
A H DiwanW J Thompson

Abstract

The flux of multisized fluorescein-isothiocyanate-labeled hydroxy ethyl starch (FITC-HES) macromolecules was used to assess changes in barrier function of rat pulmonary microvascular endothelial cell (RPMVEC) monolayers exposed to protein phosphatase (PP) inhibitors or cGMP analogs and atriopeptin (ANF). Two potent PP inhibitors, calyculin A (CalA) and okadaic acid (OA), increased RPMVEC permeability in a dose- and time-dependent manner, and CalA had a higher intrinsic activity than OA. In contrast, ANF and potent cGMP analogs had no effect on basal RPMVEC permeability. The phosphohistone PP activity contained in RPMVEC sonicates was inhibited by OA with an inhibition profile that suggested at least two components were present, with PP2A accounting for approximately 70% of the OA-inhibitable phosphohistone phosphatase activity. Following separation with heparin-Sepharose chromatography, PP activity exhibited equipotent inhibition by CalA and differential inhibition by OA. Differential inhibition of PP1 and PP2A by OA suggested that PP1 is involved in regulating RPMVEC barrier function. Permeabilized RPMVEC showed increased phosphorylation of several proteins in the presence of phosphatase inhibitors. Treatment with KT 5926, a m...Continue Reading

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Citations

Apr 16, 2013·Microvascular Research·Kyung-mi KimAlexander D Verin
Jun 1, 2005·Toxicon : Official Journal of the International Society on Toxinology·Céline Huynh-DelermeAmaury de Luze
Jul 7, 2007·Canadian Journal of Physiology and Pharmacology·Yunbo KeR John Solaro
May 25, 2004·Journal of Cellular Biochemistry·Krisztina TarAlexander D Verin
Jan 5, 2010·Vascular Pharmacology·Nagavedi S UmapathyAlexander D Verin
Jul 6, 2004·American Journal of Physiology. Lung Cellular and Molecular Physiology·Irina A KolosovaAlexander D Verin
Feb 14, 2006·Journal of Cellular Biochemistry·Krisztina TarAlexander D Verin
Aug 19, 2007·American Journal of Physiology. Lung Cellular and Molecular Physiology·Csilla CsortosAlexander D Verin
Nov 29, 2002·American Journal of Physiology. Heart and Circulatory Physiology·David R MuchaRichard C Schaeffer
Jul 17, 1999·The American Journal of Physiology·T StevensW J Thompson
Aug 16, 2000·American Journal of Physiology. Cell Physiology·T NollH M Piper
Jul 18, 2000·American Journal of Physiology. Cell Physiology·J M CarbajalR C Schaeffer
Aug 16, 2001·American Journal of Physiology. Lung Cellular and Molecular Physiology·H LumO Holian
Nov 24, 1999·The American Journal of Physiology·J M Carbajal, R C Schaeffer
Sep 10, 2003·Journal of Applied Physiology·Robert L ConhaimBruce A Harms
Dec 14, 2001·American Journal of Physiology. Lung Cellular and Molecular Physiology·Peter L GoldbergPeter A Vincent
Sep 23, 2000·American Journal of Physiology. Lung Cellular and Molecular Physiology·T M MooreT Stevens
Jun 8, 2000·The Journal of Immunology : Official Journal of the American Association of Immunologists·H E MathenyJ M Cook-Mills

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