Histamine stimulates phosphorylation of adherens junction proteins and alters their link to vimentin

Abstract

Histamine increases microvascular permeability by creating small transitory (100-400 nm) gaps between adjacent endothelial cells at sites of vascular endothelial (VE)-cadherin-based adhesion. We examined the effects of histamine on the proteins within the VE-cadherin-based adherens junction in primary human umbilical vein endothelial cells. VE-cadherin is linked not only by beta- and alpha-catenin to cortical actin but also by gamma-catenin to the intermediate filament vimentin. In mature human umbilical vein cultures, the VE-cadherin immunoprecipitate contained equivalent amounts of alpha- and beta-catenin, 130% as much beta- as gamma-catenin, and 50% as much actin as vimentin. Within 60 s, histamine decreased the fraction of VE-cadherin in the insoluble portion of the cell lysate by 35 +/- 1.5%. At the same time, histamine decreased the amount of vimentin that immunoprecipitated with VE-cadherin by 50 +/- 6%. Histamine did not affect the amount of actin or the amount of alpha-, beta-, or gamma-catenin that immunoprecipitated with VE-cadherin. Within 60 s, histamine simulated a doubling in the phosphorylation of VE-cadherin and beta- and gamma-catenin. The VE-cadherin immunoprecipitate contained kinase activity that phosphorylated VE-cadherin and gamma-catenin in vitro.

Src kinase phosphorylates vascular endothelial-cadherin in response to vascular endothelial growth factor: identification of tyrosine 685 as the unique target site

“…Tyrosine phosphorylation of VE-cadherin has been described in cells stimulated by several agents such as VEGF, thrombin, histamine, PAF and it was correlated with a rapid dissociation of adherens junctions, a critical step in angiogenesis and in inflammatory processes Zhao and Davis, 1998;Calcerrada et al, 2002;Shasby et al, 2002;Hudry-Clergeon et al, 2005). Yet, the specific tyrosine kinases involved in this process and the target sites of phosphorylation were still elusive.…”

“…Interestingly, the cytoplasmic domain of VE-cadherin contains nine tyrosine that represent potential target sites for tyrosine kinases. Indeed, VE-cadherin tyrosine phosphorylation has been reported in endothelial cells under several conditions including stimulation by VEGF Zhao and Davis, 1998;Shasby et al, 2002;Su et al, 2002). We recently reported that plateletactivating factor (PAF) increased VE-cadherin tyrosine phosphorylation in mouse endothelial cells and its association with the phosphatidylinositol 3 0 -(PtdIns3 0 )-kinase (Hudry-Clergeon et al, 2005).…”

Regulation of vimentin intermediate filaments in endothelial cells by hypoxia

“…Vimentin and keratin are the major intermediate filament proteins in endothelial cells. Endothelial barrierdisrupting agents such as histamine induce vimentin phosphorylation in HUVEC, and cause its dissociation from adhesive complexes (18). Vimentin has also been shown to regulate focal adhesions in a bone marrow endothelial cell line (23).…”

Acute regulation of tight junction ion selectivity in human airway epithelia

“…Fourth, previous studies indicated that histamine acutely increases transepithelial electrical conductance (G t ) in airway epithelia (19,22). In endothelia and cell lines, activation of histamine receptors disrupts the interaction of vascular endothelial-cadherin with the vimentin cytoskeleton to increase paracellular conductance (G p ) (23). Those studies focused on histamine's effect on adherens junctions, but whether tight junction function is also altered was not addressed.…”