title: The protein phosphatase inhibitor cantharidin alters vascular endothelial cell permeability.
abstract: In this study, we characterized the effects of the protein phosphatases type 1 (PP 1) and type 2A (PP 2A) inhibitor cantharidin in endothelial cells. We identified catalytic subunits of PP 1alpha, PP 2Aalpha, and PP 2Abeta immunologically in bovine aortic endothelial cells. Moreover, we detected mRNAs coding for catalytic subunits of PP 1alpha, PP 1beta, and PP 2Aalpha by hybridization with specific DNA probes in total RNA from these cells. Okadaic acid and cantharidin inhibited the activities of catalytic subunits of PP 1 (okadaic acid, 0.01-1 microM; cantharidin, 1-100 microM) and PP 2A (okadaic acid, 0.1 nM to 1 microM; cantharidin, 0.1-100 microM) separated by column chromatography in a concentration-dependent manner. Moreover, cantharidin (1 microM to 1 mM) increased the phosphorylation state of endothelial proteins including the regulatory light chains of myosin without affecting cytosolic calcium concentrations. Cantharidin (5-100 microM) increased the permeability of cultured endothelial cells in a time- and concentration-dependent manner. We suggest that inhibition of PP 1 and PP 2A activities by cantharidin increases endothelial permeability by enhancing the phosphorylation state of endothelial regulatory proteins. Thus, cantharidin might be a useful tool to study the function of protein phosphatases in endothelial barrier function.
authors.affiliation: Institut für Pharmakologie und Toxikologie, Westfälische Wilhelms-Universität Münster, Germany. jknapp@unimuenster.de
authors.forename: J
authors.initials: J
authors.lastname: Knapp
authors.forename: P
authors.initials: P
authors.lastname: Bokník
authors.forename: I
authors.initials: I
authors.lastname: Lüss
authors.forename: S
authors.initials: S
authors.lastname: Huke
authors.forename: B
authors.initials: B
authors.lastname: Linck
authors.forename: H
authors.initials: H
authors.lastname: Lüss
authors.forename: F U
authors.initials: FU
authors.lastname: Müller
authors.forename: T
authors.initials: T
authors.lastname: Müller
authors.forename: P
authors.initials: P
authors.lastname: Nacke
authors.forename: T
authors.initials: T
authors.lastname: Noll
authors.forename: H M
authors.initials: HM
authors.lastname: Piper
authors.forename: W
authors.initials: W
authors.lastname: Schmitz
authors.forename: U
authors.initials: U
authors.lastname: Vahlensieck
authors.forename: J
authors.initials: J
authors.lastname: Neumann
mesh_terms: D000818:Animals; D001011:Aorta; D002193:Cantharidin / Q000494:pharmacology*; D002417:Cattle; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007700:Kinetics; D018994:Myosin Light Chains / Q000302:isolation & purification / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000235:genetics*; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D004791:Enzyme Inhibitors; D018994:Myosin Light Chains; D010750:Phosphoproteins; D012333:RNA, Messenger; D012710:Serum Albumin, Bovine; D010749:Phosphoprotein Phosphatases; D002193:Cantharidin