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

Talaibek Borbiev, Alexander Dmitriyevich Verin, Shu Shi, Feng Liu, Joe G N Garcia

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

Thrombin-induced endothelial cell barrier dysfunction is tightly linked to Ca2+-dependent cytoskeletal protein reorganization. In this study, we found that thrombin increased Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) activities in a Ca2+- 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.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume280
Issue number5 25-5
StatePublished - May 1 2001
Externally publishedYes

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Calcium-Calmodulin-Dependent Protein Kinase Type 2
Calcium-Calmodulin-Dependent Protein Kinases
Thrombin
Endothelial Cells
Filamins
Phosphorylation
Electric Impedance
Myosin Light Chains
Cytoskeletal Proteins
Protein Kinase Inhibitors
Endothelium
Albumins
Permeability
Lung
Peptides

Keywords

  • Filamin
  • Myosin light chain phosphorylation
  • Thrombin
  • Transendothelial electrical resistance

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Cell Biology
  • Physiology
  • Physiology (medical)

Cite this

Regulation of endothelial cell barrier function by calcium/calmodulin-dependent protein kinase II. / Borbiev, Talaibek; Verin, Alexander Dmitriyevich; Shi, Shu; Liu, Feng; Garcia, Joe G N.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 280, No. 5 25-5, 01.05.2001.

Research output: Contribution to journalArticle

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