Regulation of reactive oxygen species-induced endothelial cell-cell and cell-matrix contacts by focal adhesion kinase and adherens junction proteins

• Published in: American journal of physiology. Lung cellular and molecular physiology 2005-12, Vol.289 (6), p.L999-L1010
• Main Authors: Usatyuk, Peter V, Natarajan, Viswanathan
• Format: Article
• Language: English
• Subjects: Actin Cytoskeleton - metabolism; Adherens Junctions - metabolism; Animals; Antigens, CD; beta Catenin - metabolism; Cadherins - metabolism; Cattle; Cell Adhesion - drug effects; Cell Adhesion - physiology; Cell Communication - drug effects; Cell Communication - physiology; Cells, Cultured; Electric Impedance; Endothelial Cells - cytology; Endothelial Cells - metabolism; Extracellular Matrix - metabolism; Focal Adhesion Protein-Tyrosine Kinases - metabolism; Hydrogen Peroxide - pharmacology; Oxidants - pharmacology; Paxillin - metabolism; Pulmonary Artery - cytology; Pulmonary Artery - metabolism; Reactive Oxygen Species - metabolism
• ISSN: 1040-0605; 1522-1504
• DOI: 10.1152/ajplung.00211.2005

Section of Pulmonary and Critical Care Medicine, University of Chicago, Chicago, Illinois Submitted 10 May 2005 ; accepted in final form 18 July 2005 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, -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, -catenin, and VE-cadherin and cell-cell adhesion. Additionally, FRNK prevented H 2 O 2 -induced distribution of FAK, paxillin, -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. focal adhesion kinase-related non-kinase; vascular endothelial-cadherin; -catenin; hydrogen peroxide; transendothelial electrical resistance Address for reprint requests and other correspondence: V. Natarajan, Dept. of Medicine, Section of Pulmonary and Critical Care Medicine, Univ. of Chicago, C/S Bldg., Rm. 408, 929 E. 57th St., Chicago, IL 60637 (e-mail: vnataraj{at}medicine.bsd.uchicago.edu )