Skip to main content

ICAM-1-mediated, Src- and Pyk2-dependent vascular endothelial cadherin tyrosine phosphorylation is required for leukocyte transendothelial migration.

Publication ,  Journal Article
Allingham, MJ; van Buul, JD; Burridge, K
Published in: J Immunol
September 15, 2007

Leukocyte transendothelial migration (TEM) has been modeled as a multistep process beginning with rolling adhesion, followed by firm adhesion, and ending with either transcellular or paracellular passage of the leukocyte across the endothelial monolayer. In the case of paracellular TEM, endothelial cell (EC) junctions are transiently disassembled to allow passage of leukocytes. Numerous lines of evidence demonstrate that tyrosine phosphorylation of adherens junction proteins, such as vascular endothelial cadherin (VE-cadherin) and beta-catenin, correlates with the disassembly of junctions. However, the role of tyrosine phosphorylation in the regulation of junctions during leukocyte TEM is not completely understood. Using human leukocytes and EC, we show that ICAM-1 engagement leads to activation of two tyrosine kinases, Src and Pyk2. Using phospho-specific Abs, we show that engagement of ICAM-1 induces phosphorylation of VE-cadherin on tyrosines 658 and 731, which correspond to the p120-catenin and beta-catenin binding sites, respectively. These phosphorylation events require the activity of both Src and Pyk2. We find that inhibition of endothelial Src with PP2 or SU6656 blocks neutrophil transmigration (71.1 +/- 3.8% and 48.6 +/- 3.8% reduction, respectively), whereas inhibition of endothelial Pyk2 also results in decreased neutrophil transmigration (25.5 +/- 6.0% reduction). Moreover, overexpression of the nonphosphorylatable Y658F or Y731F mutants of VE-cadherin impairs transmigration of neutrophils compared with overexpression of wild-type VE-cadherin (32.7 +/- 7.1% and 38.8 +/- 6.5% reduction, respectively). Our results demonstrate that engagement of ICAM-1 by leukocytes results in tyrosine phosphorylation of VE-cadherin, which is required for efficient neutrophil TEM.

Duke Scholars

Published In

J Immunol

DOI

ISSN

0022-1767

Publication Date

September 15, 2007

Volume

179

Issue

6

Start / End Page

4053 / 4064

Location

United States

Related Subject Headings

  • src-Family Kinases
  • beta Catenin
  • Up-Regulation
  • Tyrosine
  • Tumor Necrosis Factor-alpha
  • Signal Transduction
  • Protein Binding
  • Phosphorylation
  • Phosphoproteins
  • Leukocytes
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Allingham, M. J., van Buul, J. D., & Burridge, K. (2007). ICAM-1-mediated, Src- and Pyk2-dependent vascular endothelial cadherin tyrosine phosphorylation is required for leukocyte transendothelial migration. J Immunol, 179(6), 4053–4064. https://doi.org/10.4049/jimmunol.179.6.4053
Allingham, Michael J., Jaap D. van Buul, and Keith Burridge. “ICAM-1-mediated, Src- and Pyk2-dependent vascular endothelial cadherin tyrosine phosphorylation is required for leukocyte transendothelial migration.J Immunol 179, no. 6 (September 15, 2007): 4053–64. https://doi.org/10.4049/jimmunol.179.6.4053.
Allingham, Michael J., et al. “ICAM-1-mediated, Src- and Pyk2-dependent vascular endothelial cadherin tyrosine phosphorylation is required for leukocyte transendothelial migration.J Immunol, vol. 179, no. 6, Sept. 2007, pp. 4053–64. Pubmed, doi:10.4049/jimmunol.179.6.4053.

Published In

J Immunol

DOI

ISSN

0022-1767

Publication Date

September 15, 2007

Volume

179

Issue

6

Start / End Page

4053 / 4064

Location

United States

Related Subject Headings

  • src-Family Kinases
  • beta Catenin
  • Up-Regulation
  • Tyrosine
  • Tumor Necrosis Factor-alpha
  • Signal Transduction
  • Protein Binding
  • Phosphorylation
  • Phosphoproteins
  • Leukocytes