title: H(2)O(2)-mediated permeability II: importance of tyrosine phosphatase and kinase activity.
abstract: We previously reported that exposure of endothelial cells to H(2)O(2) results in a loss of cell-cell apposition and increased endothelial solute permeability. The purpose of this study was to determine how tyrosine phosphorylation and tyrosine phosphatases contribute to oxidant-mediated disorganization of endothelial cell junctions. We found that H(2)O(2) caused a rapid decrease in total cellular phosphatase activity that facilitates a compensatory increase in cellular phosphotyrosine residues. H(2)O(2) exposure also results in increased endothelial monolayer permeability, which was attenuated by pp60, an inhibitor of src kinase. Inhibition of protein tyrosine phosphatase activity by phenylarsine oxide (PAO) demonstrated a similar permeability profile compared with H(2)O(2), suggesting that tyrosine phosphatase activity is important in maintaining a normal endothelial solute barrier. Immunofluorescence shows that H(2)O(2) exposure caused a loss of pan-reactive cadherin and beta-catenin from cell junctions that was not blocked by the src kinase inhibitor PP1. H(2)O(2) also caused beta-catenin to dissociate from the endothelial cytoskeleton, which was not prevented by PP1. Finally, we determined that PP1 did not prevent cadherin internalization. These data suggest that oxidants like H(2)O(2) produce biological effects through protein phosphotyrosine modifications by decreasing total cellular phosphatase activity combined with increased src kinase activity, resulting in increased endothelial solute permeability.
authors.affiliation: Department of Genomics and Pathobiology, University of Alabama Birmingham, 35294, USA.
authors.forename: C G
authors.initials: CG
authors.lastname: Kevil
authors.forename: N
authors.initials: N
authors.lastname: Okayama
authors.forename: J S
authors.initials: JS
authors.lastname: Alexander
mesh_terms: D000818:Animals; D001152:Arsenicals / Q000494:pharmacology; D015820:Cadherins / Q000378:metabolism; D002417:Cattle; D002463:Cell Membrane Permeability / Q000502:physiology*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D016877:Oxidants / Q000494:pharmacology; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism*; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011651:Pulmonary Artery / Q000033:anatomy & histology; D015534:Trans-Activators; D051176:beta Catenin; D019061:src-Family Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D001152:Arsenicals; D015820:Cadherins; D003598:Cytoskeletal Proteins; D004791:Enzyme Inhibitors; D016877:Oxidants; D015534:Trans-Activators; D051176:beta Catenin; C029341:oxophenylarsine; D019000:Phosphotyrosine; D006861:Hydrogen Peroxide; D019061:src-Family Kinases; D017027:Protein Tyrosine Phosphatases