Regulation of reactive oxygen species-induced endothelial cell-cell and cell-matrix contacts by focal adhesion kinase and adherens junction proteins.
Oxidants, generated by activated neutrophils, have been implicated in the pathophysiology of vascular disorders and lung injury; however, mechanisms of oxidant-mediated endothelial barrier dysfunction are unclear. Here, we have investigated the role of focal adhesion kinase (FAK) in regulating hydrogen peroxide (H(2)O(2))-mediated tyrosine phosphorylation of intercellular adhesion proteins and barrier function in endothelium. Treatment of bovine pulmonary artery endothelial cells (BPAECs) with H(2)O(2) increased tyrosine phosphorylation of FAK, paxillin, beta-catenin, and vascular endothelial (VE)-cadherin and decreased transendothelial electrical resistance (TER), an index of cell-cell adhesion and/or cell-matrix adhesion. To study the role of FAK in H(2)O(2)-induced TER changes, BPAECs were transfected with vector or FAK wild-type or FAK-related non-kinase (FRNK) plasmids. Overexpression of FRNK reduced FAK expression and attenuated H(2)O(2)-mediated tyrosine phosphorylation of FAK, paxillin, beta-catenin, and VE-cadherin and cell-cell adhesion. Additionally, FRNK prevented H(2)O(2)-induced distribution of FAK, paxillin, beta-catenin, or VE-cadherin toward focal adhesions and cell-cell adhesions but not actin stress fiber formation. These results suggest that activation of FAK by H(2)O(2) is an important event in oxidant-mediated VE barrier function regulated by cell-cell and cell-matrix contacts.
Usatyuk
Peter V
PV
Dept. of Medicine, Section of Pulmonary and Critical Care Medicine, University of Chicago, C/S Bldg., Rm. 408, 929 E. 57th Street, Chicago, IL 60637, USA.
Natarajan
Viswanathan
V
D008841:Actin Cytoskeleton / Q000378:metabolism; D022005:Adherens Junctions / Q000378:metabolism*; D000818:Animals; D015703:Antigens, CD; D015820:Cadherins / Q000378:metabolism; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002450:Cell Communication / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D017097:Electric Impedance; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D016877:Oxidants / Q000494:pharmacology; D051419:Paxillin / Q000378:metabolism; D011651:Pulmonary Artery / Q000166:cytology / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism; D000099152:Cadherin 5
D015703:Antigens, CD; D015820:Cadherins; D051416:Focal Adhesion Protein-Tyrosine Kinases; D006861:Hydrogen Peroxide; D016877:Oxidants; D051419:Paxillin; D017382:Reactive Oxygen Species; D051176:beta Catenin; D000099152:Cadherin 5