Diperoxovanadate alters endothelial cell focal contacts and barrier function: Role of tyrosine phosphorylation

Joe G N Garcia, Kane L. Schaphorst, Alexander Dmitriyevich Verin, Suryanarayana Vepa, Carolyn E. Patterson, Viswanathan Natarajan

Research output: Contribution to journalArticle

63 Scopus citations

Abstract

Diperoxovanadate (DPV), a potent tyrosine kinase activator and protein tyrosine phosphatase inhibitor, was utilized to explore bovine pulmonary artery endothelial cell barrier regulation. DPV produced dose-dependent decreases in transendothelial electrical resistance (TER) and increases in permeability to albumin, which were preceded by brief increases in TER (peak TER effect at 10-15 min). The significant and sustained DPV-mediated TER reductions were primarily the result of decreased intercellular resistance, rather than decreased resistance between the cell and the extracellular matrix, and were reduced by pretreatment with the tyrosine kinase inhibitor genistein but not by inhibition of p42/p44 mitogen-activating protein kinases. Immunofluorescent analysis after DPV challenge revealed dramatic F-actin polymerization and stress-fiber assembly and increased colocalization of tyrosine phosphoproteins with F-actin in a circumferential pattern at the cell periphery, changes that were abolished by genistein. The phosphorylation of focal adhesion and adherens junction proteins on tyrosine residues was confirmed in immunoprecipitates of focal adhesion kinase and cadherin-associated proteins in which dramatic dose-dependent tyrosine phosphorylation was observed after DPV stimulation. We speculate that DPV enhances endothelial cell monolayer integrity via focal adhesion plaque phosphorylation and produces subsequent monolayer destabilization of adherens junctions initiated by adherens junction protein tyrosine phosphorylation catalyzed by p60(src) or Src-related tyrosine kinases.

Original languageEnglish (US)
Pages (from-to)2333-2343
Number of pages11
JournalJournal of Applied Physiology
Volume89
Issue number6
Publication statusPublished - Dec 20 2000
Externally publishedYes

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Keywords

  • Adherens junctions
  • Cadherin
  • Catenin
  • Electrical resistance

ASJC Scopus subject areas

  • Physiology
  • Endocrinology
  • Orthopedics and Sports Medicine
  • Physical Therapy, Sports Therapy and Rehabilitation

Cite this

Garcia, J. G. N., Schaphorst, K. L., Verin, A. D., Vepa, S., Patterson, C. E., & Natarajan, V. (2000). Diperoxovanadate alters endothelial cell focal contacts and barrier function: Role of tyrosine phosphorylation. Journal of Applied Physiology, 89(6), 2333-2343.