SHP2 association with VE-cadherin complexes in human endothelial cells is regulated by thrombin.
Thrombin-mediated changes in endothelial cell adherens junctions modulate vascular permeability. We demonstrate that the nonreceptor protein-tyrosine phosphatase SHP2 co-precipitates with VE-cadherin complexes in confluent, quiescent human umbilical vein endothelial cells. Ligand-binding blots using a SHP2-glutathione S-transferase fusion peptide established that SHP2 associates selectively with beta-catenin in VE-cadherin complexes. Thrombin treatment of human umbilical vein endothelial cells promotes SHP2 tyrosine phosphorylation and dissociation from VE-cadherin complexes. The loss of SHP2 from the cadherin complexes correlates with a dramatic increase in the tyrosine phosphorylation of beta-catenin, gamma-catenin, and p120-catenin complexed with VE-cadherin. We propose that thrombin regulates the tyrosine phosphorylation of VE-cadherin-associated beta-catenin, gamma-catenin, and p120-catenin by modulating the quantity of SHP2 associated with VE-cadherin complexes. Such changes in adherens junction complex composition likely underlie thrombin-elicited alterations in endothelial monolayer permeability.
Ukropec
J A
JA
Department of Physiology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107-6799, USA.
Hollinger
M K
MK
Salva
S M
SM
Woolkalis
M J
MJ
D015703:Antigens, CD; D015820:Cadherins / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004730:Endothelium, Vascular / Q000201:enzymology* / Q000378:metabolism; D006490:Hemostatics / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D010766:Phosphorylation; D011233:Precipitin Tests; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology*; D013997:Time Factors; D015534:Trans-Activators; D014471:Umbilical Veins / Q000201:enzymology / Q000378:metabolism; D051176:beta Catenin; D000099152:Cadherin 5
D015703:Antigens, CD; D015820:Cadherins; D003598:Cytoskeletal Proteins; D006490:Hemostatics; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases; D011993:Recombinant Fusion Proteins; D013917:Thrombin; D015534:Trans-Activators; D051176:beta Catenin; D000099152:Cadherin 5; C495270:CTNNB1 protein, human; C516724:PTPN11 protein, human; C509249:PTPN6 protein, human