Phospholipase D2 restores endothelial barrier function by promoting PTPN14-mediated VE-cadherin dephosphorylation

J Biol Chem. 2020 May 29;295(22):7669-7685. doi: 10.1074/jbc.RA119.011801. Epub 2020 Apr 23.


Increased permeability of vascular lung tissues is a hallmark of acute lung injury and is often caused by edemagenic insults resulting in inflammation. Vascular endothelial (VE)-cadherin undergoes internalization in response to inflammatory stimuli and is recycled at cell adhesion junctions during endothelial barrier re-establishment. Here, we hypothesized that phospholipase D (PLD)-generated phosphatidic acid (PA) signaling regulates VE-cadherin recycling and promotes endothelial barrier recovery by dephosphorylating VE-cadherin. Genetic deletion of PLD2 impaired recovery from protease-activated receptor-1-activating peptide (PAR-1-AP)-induced lung vascular permeability and potentiated inflammation in vivo In human lung microvascular endothelial cells (HLMVECs), inhibition or deletion of PLD2, but not of PLD1, delayed endothelial barrier recovery after thrombin stimulation. Thrombin stimulation of HLMVECs increased co-localization of PLD2-generated PA and VE-cadherin at cell-cell adhesion junctions. Inhibition of PLD2 activity resulted in prolonged phosphorylation of Tyr-658 in VE-cadherin during the recovery phase 3 h post-thrombin challenge. Immunoprecipitation experiments revealed that after HLMVECs are thrombin stimulated, PLD2, VE-cadherin, and protein-tyrosine phosphatase nonreceptor type 14 (PTPN14), a PLD2-dependent protein-tyrosine phosphatase, strongly associate with each other. PTPN14 depletion delayed VE-cadherin dephosphorylation, reannealing of adherens junctions, and barrier function recovery. PLD2 inhibition attenuated PTPN14 activity and reversed PTPN14-dependent VE-cadherin dephosphorylation after thrombin stimulation. Our findings indicate that PLD2 promotes PTPN14-mediated dephosphorylation of VE-cadherin and that redistribution of VE-cadherin at adherens junctions is essential for recovery of endothelial barrier function after an edemagenic insult.

Keywords: VE-cadherin; cadherin; endothelial barrier restoration; endothelial cell; endothelial dysfunction; lipid signaling; phosphatidic acid signaling; phospholipase D2 (PLD2); phosphotyrosine signaling; protein tyrosine phosphatase non-receptor type 14 (PTPN14).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adherens Junctions / metabolism
  • Animals
  • Antigens, CD / metabolism*
  • Blood-Air Barrier / cytology
  • Blood-Air Barrier / metabolism*
  • Cadherins / metabolism*
  • Endothelial Cells / cytology
  • Endothelial Cells / metabolism*
  • Female
  • Humans
  • Male
  • Mice
  • Phospholipase D / metabolism*
  • Phosphorylation / drug effects
  • Protein Tyrosine Phosphatases, Non-Receptor / metabolism*
  • Thrombin / pharmacology


  • Antigens, CD
  • Cadherins
  • cadherin 5
  • PTPN14 protein, human
  • PTPN14 protein, mouse
  • Protein Tyrosine Phosphatases, Non-Receptor
  • phospholipase D2
  • Phospholipase D
  • Thrombin