title: c-Abl mediated tyrosine phosphorylation of paxillin regulates LPS-induced endothelial dysfunction and lung injury.
abstract: Paxillin is phosphorylated at multiple residues; however, the role of tyrosine phosphorylation of paxillin in endothelial barrier dysfunction and acute lung injury (ALI) remains unclear. We used siRNA and site-specific nonphosphorylable mutants of paxillin to abrogate the function of paxillin to determine its role in lung endothelial permeability and ALI. In vitro, lipopolysaccharide (LPS) challenge of human lung microvascular endothelial cells (HLMVECs) resulted in enhanced tyrosine phosphorylation of paxillin at Y31 and Y118 with no significant change in Y181 and significant barrier dysfunction. Knockdown of paxillin with siRNA attenuated LPS-induced endothelial barrier dysfunction and destabilization of VE-cadherin. LPS-induced paxillin phosphorylation at Y31 and Y118 was mediated by c-Abl tyrosine kinase, but not by Src and focal adhesion kinase. c-Abl siRNA significantly reduced LPS-induced endothelial barrier dysfunction. Transfection of HLMVECs with paxillin Y31F, Y118F, and Y31/118F double mutants mitigated LPS-induced barrier dysfunction and VE-cadherin destabilization. In vivo, the c-Abl inhibitor AG957 attenuated LPS-induced pulmonary permeability in mice. Together, these results suggest that c-Abl mediated tyrosine phosphorylation of paxillin at Y31 and Y118 regulates LPS-mediated pulmonary vascular permeability and injury.
authors: Department of Pharmacology, University of Illinois, Chicago, Illinois; pfu@uic.edu.
authors: Panfeng
authors: P
authors: Fu
authors: Department of Pharmacology, University of Illinois, Chicago, Illinois;
authors: Peter V
authors: PV
authors: Usatyuk
authors: Department of Pharmacology, University of Illinois, Chicago, Illinois;
authors: Abhishek
authors: A
authors: Lele
authors: Department of Pediatrics, University of Illinois, Chicago, Illinois;
authors: Anantha
authors: A
authors: Harijith
authors: Department of Cellular and Molecular Medicine, The University of Arizona College of Medicine, Tucson, Arizona;
authors: Carol C
authors: CC
authors: Gregorio
authors: Department of Medicine, The University of Arizona College of Medicine, Tucson, Arizona; and.
authors: Joe G N
authors: JG
authors: Garcia
authors: Department of Medicine, University of Chicago, Chicago, Illinois.
authors: Ravi
authors: R
authors: Salgia
authors: Department of Pharmacology, University of Illinois, Chicago, Illinois; Department of Medicine, College of Medicine, University of Illinois, Chicago, Illinois;
authors: Viswanathan
authors: V
authors: Natarajan
mesh_terms: D055371:Acute Lung Injury / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D000818:Animals; D015703:Antigens, CD; D015820:Cadherins; D002199:Capillary Permeability / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000378:metabolism* / Q000473:pathology; D051417:Focal Adhesion Kinase 1 / Q000235:genetics / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D008070:Lipopolysaccharides / Q000633:toxicity*; D008297:Male; D051379:Mice; D051419:Paxillin / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation / Q000235:genetics; D016315:Proto-Oncogene Proteins c-abl / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D020032:Tyrphostins / Q000494:pharmacology; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D051417:Focal Adhesion Kinase 1; D008070:Lipopolysaccharides; D051419:Paxillin; D016315:Proto-Oncogene Proteins c-abl; D020032:Tyrphostins; D000099152:Cadherin 5; C496061:PXN protein, human; C496062:Pxn protein, mouse; C104965:tyrphostin AG957; C496050:PTK2 protein, human; C496051:Ptk2 protein, mouse
keywords: c-Abl; cytoskeleton; endothelial dysfunction; lipopolysaccharide; paxillin; permeability; tyrosine kinase