Regulation of endothelial cell barrier function by calcium/calmodulin-dependent protein kinase II

Am J Physiol Lung Cell Mol Physiol. 2001 May;280(5):L983-90. doi: 10.1152/ajplung.2001.280.5.L983.

Abstract

Thrombin-induced endothelial cell barrier dysfunction is tightly linked to Ca(2+)-dependent cytoskeletal protein reorganization. In this study, we found that thrombin increased Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) activities in a Ca(2+)- and time-dependent manner in bovine pulmonary endothelium with maximal activity at 5 min. Pretreatment with KN-93, a specific CaM kinase II inhibitor, attenuated both thrombin-induced increases in monolayer permeability to albumin and decreases in transendothelial electrical resistance (TER). We next explored potential thrombin-induced CaM kinase II cytoskeletal targets and found that thrombin causes translocation and significant phosphorylation of nonmuscle filamin (ABP-280), which was attenuated by KN-93, whereas thrombin-induced myosin light chain phosphorylation was unaffected. Furthermore, a cell-permeable N-myristoylated synthetic filamin peptide (containing the COOH-terminal CaM kinase II phosphorylation site) attenuated both thrombin-induced filamin phosphorylation and decreases in TER. Together, these studies indicate that CaM kinase II activation and filamin phosphorylation may participate in thrombin-induced cytoskeletal reorganization and endothelial barrier dysfunction.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Benzylamines / pharmacology
  • Calcium / metabolism
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Calcium-Calmodulin-Dependent Protein Kinases / antagonists & inhibitors
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism*
  • Capillary Permeability / drug effects
  • Capillary Permeability / physiology*
  • Cattle
  • Cell Line
  • Contractile Proteins / metabolism
  • Cytoskeleton / metabolism
  • Electric Impedance
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / metabolism*
  • Enzyme Inhibitors / pharmacology
  • Filamins
  • Microfilament Proteins / metabolism
  • Myosin Light Chains / metabolism
  • Phosphorylation / drug effects
  • Protein Transport / drug effects
  • Pulmonary Artery
  • Subcellular Fractions / metabolism
  • Sulfonamides / pharmacology
  • Thrombin / pharmacology

Substances

  • Benzylamines
  • Contractile Proteins
  • Enzyme Inhibitors
  • Filamins
  • Microfilament Proteins
  • Myosin Light Chains
  • Sulfonamides
  • KN 93
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Thrombin
  • Calcium